Researcher basic information

• 獣医学博士, 北海道大学

• https://researchmap.jp/prion?lang=en

• https://jglobal.jst.go.jp/detail?JGLOBAL_ID=201301012090071059
• http://kaken.nii.ac.jp/d/r/30219216.ja.html

• 201301012090071059

• Campylobacter
• Comprehensive gene expression analysis
• プリオン
• スクレイピー
• BSE
• 伝達性海綿状脳症
• PrP
• アストロサイト
• 神経変性
• 構造転換
• 合成ペプチド
• PrPSc
• 猫パルボウイルス亜群
• モンゴル
• 再生医療
• バイエル氏板
• 走化性
• 消化管付髄リンパ装置
• 濾胞樹状細胞
• モノクロナール抗体
• 神経細胞死
• 骨髄由来間葉系幹細胞
• 分子進化
• MEV
• 遺伝子型
• ALYマウス
• microglia
• siRNA
• PCR
• FPLV
• Neuro2a

• Life Science, Veterinary medical science
• Nanotechnology/Materials, Chemistry and chemical methodology of biomolecules
• Life Science, Virology
• Nanotechnology/Materials, Polymer chemistry

• Bachelor's degree program, School of Veterinary Medicine
• Doctoral (PhD) degree program, Graduate School of Infectious Diseases

Research activity information

• Strain Traits of Intracranially Administered L-Type Bovine Spongiform Encephalopathy Prions Are not Significantly Modified During Intraspecies Transmission in Cynomolgus Monkeys.
  Ken'ichi Hagiwara; Hiroaki Shibata; Minoru Tobiume; Yuko Sato; Keiko Ohto; Sachi Okabayashi; Nozomi Nakano; Motohiro Horiuchi; Fumiko Ono
  Microbiology and immunology, 19 Jan. 2026, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Among the three prion strains of bovine spongiform encephalopathy (BSE), classical BSE (C-BSE) prions are known causative agents of variant Creutzfeldt-Jakob disease. By contrast, human infections with L-type (L-) or H-type (H-) BSE prions have not been reported. Nonetheless, the zoonotic potential of L-BSE prions is supported by their successful primary transmission from cattle to cynomolgus macaque (Macaca fascicularis) monkeys via intracranial challenge. To assess whether the defining strain traits of L-BSE prions remain stable following secondary intraspecies transmission, we prepared brain homogenates from a diseased macaque that had previously undergone primary transmission of L-BSE prions, and intracranially administered them into two naïve macaques. Both animals succumbed to the disease within humane endpoints comparable to those observed in the primary transmission. Histopathological and immunohistochemical analyses of brain tissues showed no significant changes relative to primary transmission, including severe vacuolation and fine synaptic distribution of disease-associated forms of prion protein (PrPSc) in the cerebrum, and sparse PrPSc plaques in the cerebellum. In bioassays using C57BL/6 J mice, cattle-derived L-BSE prions and those passaged once or twice in macaques failed to transmit to mice, whereas cattle-derived C-BSE prions and their macaque-passaged counterparts were transmissible. These findings refine our understanding of L-BSE pathogenesis and confirm the stability of L-BSE prions following intracranial transmission in a nonhuman primate model.
• Antimicrobial Susceptibility of Campylobacter spp. Isolated from Cattle in Mongolia.
  Erdenebat Bulgan; Zolzaya Byambajav; Batsukh Naranchimeg; Batsaikhan Chantsal; Tsognemekh Bolormaa; Badrakh Sandagdorj; Purevdorj Nyam-Osor; Eisaku Kikuchi; Akio Suzuki; Jirachaya Toyting-Hiraishi; Toyotaka Sato; Motohiro Horiuchi
  Veterinary sciences, 12, 10, 24 Sep. 2025, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal, Poultry and cattle are the major reservoirs of Campylobacter infection in humans. However, no information is available on Campylobacter spp. in cattle in Mongolia. Thus, this study aimed to assess their prevalence and antimicrobial resistance. Between 2019 and 2023, rectal swabs were collected from cattle on dairy farms around Ulaanbaatar city and in total, 35 Campylobacter spp., including 23 C. jejuni, 7 C. hyointestinalis, 4 C. fetus, and 1 C. lari, were isolated. Multilocus sequence typing of C. jejuni cattle isolates revealed substantial genetic diversity and identified 7 sequence types (STs) including ST61, which is known to be associated with cattle and sheep. Interestingly, the antimicrobial resistance patterns of the C. jejuni cattle isolates completely differed from those of previously reported chicken isolates. Excluding one ciprofloxacin-resistant isolate, all isolates were susceptible to tetracycline and ciprofloxacin. This is the first report on the characterization of Campylobacter spp. in cattle in Mongolia. Although no official statistics of human campylobacteriosis are currently available in Mongolia, data on Campylobacter spp. in food-producing animals represent valuable information for investigating potential sources and infection routes to humans.
• Potential human health risk of carbapenem-non-susceptible Pseudomonas aeruginosa from companion animals.
  Jirachaya Toyting-Hiraishi; Toyotaka Sato; Mana Tohyama; Taro Fujino; Kaho Okada; Kazuyoshi Sasaoka; Nozomu Yokoyama; Kana Torii; Akio Suzuki; Yuzo Tsuyuki; Kensuke Nakamura; Mitsuyoshi Takiguchi; Motohiro Horiuchi
  The Journal of antimicrobial chemotherapy, 19 Sep. 2025, [Peer-reviewed], [Last author], [International Magazine]
  English, Scientific journal, BACKGROUND AND OBJECTIVES: The close bond between companion animals and humans may accelerate the spread of antimicrobial-resistant bacteria. Pseudomonas aeruginosa, an opportunistic pathogen in both, poses a public health threat due to antimicrobial resistance (AMR) and diverse virulence factors. However, One Health-based comparison remains limited. This study investigated the current AMR status and molecular characteristics of P. aeruginosa in companion animals in Japan to assess potential human health risks. METHODS: We examined 197 P. aeruginosa clinical isolates from companion animals [dogs (n = 99) and cats (n = 98)] across Japan in 2024. Antimicrobial susceptibility to human clinical antibiotics was evaluated. In carbapenem-non-susceptible isolates, multilocus sequence typing and detection of resistance genes and virulence factors were performed. RESULTS: Ciprofloxacin (20.3%) and piperacillin (10.7%) showed the highest resistance rates, with 5.6% of isolates being multidrug-resistant. Carbapenem resistance rates were 6.1% for imipenem and 1.0% for meropenem. Thirty-five isolates (17.8%) exhibited carbapenem non-susceptibility but remained susceptible to cefepime, ciprofloxacin, or amikacin. Of 27 identified sequence types, 20 (77.1% of carbapenem-non-susceptible isolates) were known in humans, including two high-risk clones (ST233 and ST298; 8.6%) reported for the first time in Japanese companion animals. These isolates carried mutations in efflux pump-related genes and multiple virulence factors. One showed close genetic relatedness to a human isolate, suggesting possible interspecies transmission. CONCLUSIONS: Our findings highlight the potential cross-species transmission risk of antimicrobial-resistant P. aeruginosa. Identification of shared high-risk clones with multiple virulence factors emphasizes the need for continuous vigilance and actions within the One Health framework.
• Design, Synthesis, and Biological Evaluation of Lipid-Modified Derivatives of Tunicamycins.
  Kazuki Yamamoto; Toyotaka Sato; Ellene H Mashalidis; Seok-Yong Lee; Motohiro Horiuchi; Satoshi Ichikawa
  Chemistry (Weinheim an der Bergstrasse, Germany), e02296, 15 Sep. 2025, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Naturally occurring antibiotic tunicamycin targets bacterial peptidoglycan biosynthesis by inhibiting bacterial phospho-N-acetylmuraminic acid (MurNAc)-pentapeptide translocase (MraY), but it also inhibits human UDP-N-acetylglucosamine (GlcNAc): polyprenol phosphate translocase (GPT), leading to cytotoxicity. This study thoroughly investigated the structure-activity relationship of the fatty acyl side chain of tunicamycin to develop MraY-selective inhibitors, based on structural differences between MraY and GPT. Longer alkyl chains and flexible structures were found to favor MraY inhibitory activity, and benzene rings were acceptable for binding. The hybrid analogue containing oleoyl group, which contributed most significantly to MraY inhibitory activity, and the MurNAc moiety, which is important for MraY-selective inhibition, showed enhanced MraY inhibitory activity as well as improved antibacterial activity against S. aureus and E. faecium.
• Unveiling the Genetic Diversity and Antimicrobial Resistance Profiles of Salmonella Population From 2016 to 2020 in Thai Canal Water.
  Jirachaya Toyting-Hiraishi; Toyotaka Sato; Neunghatai Supha; Yuwanda Thongpanich; Motohiro Horiuchi; Jeewan Thapa; Chie Nakajima; Yasuhiko Suzuki; Fuangfa Utrarachkij
  Environmental microbiology reports, 17, 4, e70160, Aug. 2025, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Salmonella is one of the important pathogens causing acute gastroenteritis, and antimicrobial-resistant Salmonella raises a critical public health concern. Canals in Bangkok, Thailand, play a vital role as sources of agricultural and daily water usage. By employing whole genome sequencing to analyse 351 Salmonella genomes isolated between 2016 and 2020, we expanded the understanding of the characteristics and antimicrobial resistance properties of Salmonella enterica found in Bangkok canals, an underrepresented biome in research. Salmonella Agona was the dominant serotype, while S. Typhimurium and its monophasic variant were periodically found. Seven new sequence types (STs) were identified, including STs 11,346, 11,347, 11,348, 11,349, 11,350, 11,351, and 11,352. Seven chromosomal-mediated gene mutations and 50 antimicrobial resistance genes were detected. The three most common resistance genes were tet(A), blaTEM-1B, and qnrS1. The tet(X4) was first identified in the Salmonella population in Thailand, and mcr-3.1 was also detected. In total, 39.0% of the strains were potentially multidrug-resistant. The strains carried double amino acid substitutions in GyrA and ParC, and a strain with GyrA substitutions and qnrS1 exhibited the strongest resistance to nalidixic acid and ciprofloxacin. Most of the ceftazidime, ceftriaxone, and cefotaxime-resistant strains (66.7%) harboured blaCTX-M-55. Col(pHAD28) was the predominant plasmid replicon type. Phylogenetic analysis of Salmonella STs 34 and 213 from canal water and the strains from databases showed the possibility of circulation of STs 34 and 213 between canal water and humans in Thailand and worldwide. These findings shed light on the circulation of antimicrobial-resistant pathogens in the environmental water and advocate for incorporating environmental sampling into comprehensive AMR surveillance programmes within a One Health framework.
• Seasonal changes in the yield and composition of camel milk in Mongolia
  Bolormaa Tsognemekh; Ganzorig Sumiya; Takai Shinji; Horiuchi Motohiro; Nyam-Osor Purevdorj
  PASTORALISM-RESEARCH POLICY AND PRACTICE, 15, 17 Jul. 2025, [Peer-reviewed]
  English, Scientific journal
• Detection of Brucella spp. from milk by quantitative PCR as a monitoring method for brucellosis in cattle in Mongolia.
  Batsukh Naranchimeg; Batsaikhan Chantsal; Badrakh Sandagdorj; Tsognemekh Bolormaa; Purevdorj Ulzii-Orshikh; Adilbish Altanchimeg; Sumiya Ganzorig; Vanaabaatar Batbaatar; Motohiro Horiuchi; Purevdorj Nyam-Osor
  BMC veterinary research, 21, 1, 466, 466, 15 Jul. 2025, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal, BACKGROUND: Brucellosis in livestock is endemic in Mongolia, and efficient monitoring is required for clarifying its prevalence. Milk can be obtained noninvasively and is useful for monitoring brucellosis in livestock. However, the usefulness of milk in monitoring brucellosis should be clarified. MATERIALS AND METHODS: Serum and milk samples were obtained from 326 cows from six farms near Ulaanbaatar City between 2020 and 2022. Serum and milk were assessed using the Rose Bengal Test (RBT) and Milk Ring Test (MRT), respectively. All milk samples were further subjected to bacterial isolation and DNA extraction. DNA samples were analyzed with quantitative PCR (qPCR) targeting Brucella genus-specific IS711 insertion sequence to detect and estimate Brucella spp. levels in milk. qPCR-positive samples were further subjected to single nucleotide polymorphism (SNP) assay to discriminate B. abortus field strains from the S19 vaccine strain. RESULTS: Of the 326 milk samples, 108 (33.1%) were revealed to be positive for Brucella spp. by qPCR, whereas only 5 samples (1.5%) were deemed positive via the bacterial isolation. No S19 vaccine strain was identified in the IS711 qPCR-positive milk samples by the SNP assay. Although qPCR detected Brucella spp. from milk, which was obtained from cows in three lactation stages, the detection ratio was significantly higher in the early lactation stage than in the middle lactation stage. Additionally, the five milk samples from which Brucella spp, were isolated exhibited the top 5 estimated colony forming units among the IS711 qPCR-positive samples, indicating that detection sensitivity of the IS711 qPCR is extremely higher than that of bacterial culture. There was a tendency that milk samples from RBT- and MRT-positive cows are more likely to be positive by IS711 qPCR. CONCLUSION: The results revealed that analysis of milk with qPCR is easy and sensitive monitoring method for detecting Brucella infection in livestock.
• 国境を越えて:留学生・ポスドクの挑戦 2016年から2020年までのタイの運河水におけるSalmonellaの遺伝的多様性と抗菌薬耐性(Beyond Borders: Challenges for International Students and Postdocs Genetic diversity and antimicrobial resistance of Salmonella in Thai canal water from 2016 to 2020)
  Toyting Jirachaya; 佐藤 豊孝; Supha Neunghatai; Thongpanich Yuwanda; 堀内 基広; Thapa Jeewan; 中島 千絵; 鈴木 定彦; Utrarachkij Fuangfa
  日本細菌学雑誌, 80, 2, 34, 34, 日本細菌学会, Apr. 2025
  English
• Rapid and Integrated Bacterial Evolution Analysis unveils gene mutations and clinical risk of Klebsiella pneumoniae.
  Kojiro Uemura; Toyotaka Sato; Soh Yamamoto; Noriko Ogasawara; Jirachaya Toyting; Kotaro Aoki; Akira Takasawa; Masayuki Koyama; Atsushi Saito; Takayuki Wada; Kaho Okada; Yurie Yoshida; Koji Kuronuma; Chie Nakajima; Yasuhiko Suzuki; Motohiro Horiuchi; Kenichi Takano; Satoshi Takahashi; Hirofumi Chiba; Shin-Ichi Yokota
  Nature communications, 16, 1, 2917, 2917, 25 Mar. 2025, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Bacteria continually evolve. Previous studies have evaluated bacterial evolution in retrospect, but this approach is based on only speculation. Cohort studies are reliable but require a long duration. Additionally, identifying which genetic mutations that have emerged during bacterial evolution possess functions of interest to researchers is an exceptionally challenging task. Here, we establish a Rapid and Integrated Bacterial Evolution Analysis (RIBEA) based on serial passaging experiments using hypermutable strains, whole-genome and transposon-directed sequencing, and in vivo evaluations to monitor bacterial evolution in a cohort for one month. RIBEA reveals bacterial factors contributing to serum and antimicrobial resistance by identifying gene mutations that occurred during evolution in the major respiratory pathogen Klebsiella pneumoniae. RIBEA also enables the evaluation of the risk for the progression and the development of invasive ability from the lung to blood and antimicrobial resistance. Our results demonstrate that RIBEA enables the observation of bacterial evolution and the prediction and identification of clinically relevant high-risk bacterial strains, clarifying the associated pathogenicity and the development of antimicrobial resistance at genetic mutation level.
• Prevalence, antimicrobial susceptibility, and virulence profiles of fluoroquinolone-resistant <i>Escherichia coli</i> isolated from companion animals in Sapporo, Japan
  Aiko MAEDA; Toyotaka SATO; Jirachaya TOYTING-HIRAISHI; Akio SUZUKI; Yuuji HOSHINO; Shingo TORIGOE; Keiichiro SAKAKIBARA; Satoshi TAMAI; Tooru TACHIBANA; Motohiro HORIUCHI
  Journal of Veterinary Medical Science, Japanese Society of Veterinary Science, 2025, [Peer-reviewed], [Last author]
  Scientific journal
• Lack of Evidence for Transmission of Atypical H-Type Bovine Spongiform Encephalopathy Prions (H-BSE Prions) by Intracranial and Oral Challenges to Nonhuman Primates.
  Hiroaki Shibata; Fumiko Ono; Yuko Sato; Keiko Ohto; Nozomi Nakano; Morikazu Imamura; Motohiro Horiuchi; Minoru Tobiume; Ken'ichi Hagiwara
  Microbiology and immunology, 69, 1, 25, 34, Jan. 2025, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Bovine spongiform encephalopathy (BSE) is a prion disease in cattle caused by classical-type (C-), L-type (L-), or H-type (H-) BSE prions. While C-BSE prions are zoonotic agents responsible for variant Creutzfeldt-Jakob disease, L- and H-BSE prions are believed not to be connected to human prion diseases. However, L-BSE prions have been shown to transmit to cynomolgus monkeys (Macaca fascicularis), suggesting they may have zoonotic potential. In the present study, we examined whether H-BSE prions are transmissible to cynomolgus monkeys. The monkeys were injected intracranially (n = 2) or given orally (n = 2) with brain homogenates from a cow infected with H-BSE prions. After asymptomatic observation periods of 4-6 years, the monkeys were euthanized for autopsy. Histological examination of the brain did not reveal any pathological changes. Immunohistochemical and Western blot analyses did not detect disease-associated forms of prion protein (PrPSc) in the brain, peripheral neurons, or lymphatic tissues. The unsuccessful transmission indicates an effective barrier against the transmission of cattle H-BSE prions to cynomolgus monkeys. Based on the results obtained in this nonhuman primate model, we estimated that the potential transmission of H-BSE prions to humans is substantially lower than C- and L-BSE prions.
• Antimicrobial resistance and self-reported hand hygiene awareness before and after an infection prevention and control programme: A 7-year analysis in a small animal veterinary teaching hospital.
  Kazuyoshi Sasaoka; Toyotaka Sato; Keitaro Morishita; Kenji Hosoya; Nozomu Yokoyama; Takachika Sato; Motohiro Horiuchi; Mitsuyoshi Takiguchi
  Veterinary journal (London, England : 1997), 306, 106154, 106154, Aug. 2024, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Infection prevention and control (IPC) in veterinary medicine is crucial to protect patients, owners, staff, and the public. An IPC programme is recommended for every animal hospital. The objective of this retrospective longitudinal study was to describe the changes in bacterial and multidrug-resistant (MDR) bacterial isolates and self-reported hand hygiene awareness and practices after an IPC programme to assess the long-term effect of this programme in small animal veterinary medicine. The IPC programme was implemented at our veterinary teaching hospital in April 2018, which included the establishment of an infection control task force, regular IPC lectures and poster campaigns, infrastructure improvement, and manual refinement. Laboratory-based surveillance was retrospectively conducted before and after the programme (January 2016-December 2022). Level and slope changes in bacterial isolates were evaluated using interrupted time-series analysis. Self-reported hand hygiene awareness and practices were assessed using an annual questionnaire. Additionally, hygiene product purchases during the study period were investigated. The monthly number of total and MDR bacterial isolates decreased significantly after the programme (MDR level change: -0.426; 95% confidence interval: -0.744, -0.109; P = 0.009; and MDR slope change: -0.035; 95% confidence interval: -0.058, -0.011; P = 0.003). Additionally, awareness of hand hygiene before touching animals improved after the programme. Overall self-reported hand hygiene practices improved, and hygiene product purchases significantly increased. These results suggested that the IPC programme may have long-term effects regarding reducing total and MDR bacterial isolates and improving hand hygiene awareness in veterinary medicine.
• Characterization of Shiga Toxin-producing Escherichia coli Isolated from Cattle Around Ulaanbaatar City, Mongolia.
  Erdenebat Bulgan; Zolzaya Byambajav; Narantuya Ayushjav; Yuji Hirai; Misaki Tanaka; Nyam-Osor Purevdorj; Sandagdorj Badrakh; Akio Suzuki; Yusuke Komatsu; Toyotaka Sato; Motohiro Horiuchi
  Journal of food protection, 87, 7, 100294, 100294, Jul. 2024, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal, Shiga toxin-producing Escherichia coli (STEC) are associated with severe infections including hemorrhagic colitis and hemolytic uremic syndrome in humans. Ruminants are known as reservoirs of STEC; however, no data are available on STEC in ruminants in Mongolia, where more than 5 million cattle and 25 million sheep are raised. To disclose the existence and characteristics of STEC in Mongolia, in this study, we isolated and characterized STEC from cattle in Mongolia. We collected 350 rectal swabs of cattle from 30 farms near Ulaanbaatar city and isolated 45 STEC from 21 farms. Rectal swabs were precultured with modified Escherichia coli broth and then inoculated to Cefixime-Tellurite Sorbitol MacConkey agar plate and/or CHROMagar STEC agar plate for the isolation of STEC. The isolation ratios in each farm were from 0% to 40%. Multiplex PCR for the estimation of O- and H-serotypes identified 12 O-genotypes (Og-types) and 11 H-genotypes (Hg-types) from 45 isolates; however, Og-types of 19 isolates could not be determined. Stx gene subtyping by PCR identified 2 stx1 subtypes (1a and 1c) and 4 stx2 subtypes (2a, 2c, 2d, and 2g). Forty-five isolates were divided into 21 different groups based on the Og- and Hg-types, stx gene subtypes and the existence of virulence factors, ehxA, eae, and saa, which includes several major serotypes associated with human illness such as O26:H11 and O157:H7. The most dominant isolate, OgUT:H19 [stx1a (+), stx2a (+), ehxA (+) and saa (+)], was isolated from eight farms. This is the first report on the characterization of STEC in cattle in Mongolia, and the results suggest the importance of further monitoring of STEC contamination in the food chains as well as STEC infection in humans.
• Development of a natural product optimization strategy for inhibitors against MraY, a promising antibacterial target.
  Kazuki Yamamoto; Toyotaka Sato; Aili Hao; Kenta Asao; Rintaro Kaguchi; Shintaro Kusaka; Radhakrishnam Raju Ruddarraju; Daichi Kazamori; Kiki Seo; Satoshi Takahashi; Motohiro Horiuchi; Shin-Ichi Yokota; Seok-Yong Lee; Satoshi Ichikawa
  Nature communications, 15, 1, 5085, 5085, 14 Jun. 2024, [Peer-reviewed], [International Magazine]
  English, Scientific journal, MraY (phospho-N-acetylmuramoyl-pentapeptide-transferase) inhibitory natural products are attractive molecules as candidates for a new class of antibacterial agents to combat antimicrobial-resistant bacteria. Structural optimization of these natural products is required to improve their drug-like properties for therapeutic use. However, chemical modifications of these natural products are painstaking tasks due to complex synthetic processes, which is a bottleneck in advancing natural products to the clinic. Here, we develop a strategy for a comprehensive in situ evaluation of the build-up library, which enables us to streamline the preparation of the analogue library and directly assess its biological activities. We apply this approach to a series of MraY inhibitory natural products. Through construction and evaluation of the 686-compound library, we identify promising analogues that exhibit potent and broad-spectrum antibacterial activity against highly drug-resistant strains in vitro as well as in vivo in an acute thigh infection model. Structures of the MraY-analogue complexes reveal distinct interaction patterns, suggesting that these analogues represent MraY inhibitors with unique binding modes. We further demonstrate the generality of our strategy by applying it to tubulin-binding natural products to modulate their tubulin polymerization activities.
• Traces of pandemic fluoroquinolone-resistant Escherichia coli clone ST131 transmitted from human society to aquatic environments and wildlife in Japan.
  Toyotaka Sato; Kojiro Uemura; Mitsuru Yasuda; Aiko Maeda; Toshifumi Minamoto; Kazuki Harada; Michiyo Sugiyama; Shiori Ikushima; Shin-Ichi Yokota; Motohiro Horiuchi; Satoshi Takahashi; Testuo Asai
  One health (Amsterdam, Netherlands), 18, 100715, 100715, Jun. 2024, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Transmission of antimicrobial-resistant bacteria among humans, animals, and the environment is a growing concern worldwide. The distribution of an international high-risk fluoroquinolone-resistant Escherichia coli clone, ST131, has been documented in clinical settings. However, the transmission of ST131 from humans to surrounding environments remains poorly elucidated. To comprehend the current situation and identify the source of ST131 in nature, we analyzed the genetic features of ST131 isolates from the aquatic environment (lake/river water) and wildlife (fox, raccoon, raccoon dog, and deer) and compared them with the features of isolates from humans in Japan using accessory and core genome single nucleotide polymorphism (SNP) analyses. We identified ST131 isolates belonging to the same phylotype and genome clusters (four of eight clusters were concomitant) with low SNP distance between the human isolates and those from the aquatic environment and wildlife. These findings warn of ST131 transmission between humans and the surrounding environment in Japan.
• Excitatory neuron-prone prion propagation and excitatory neuronal loss in prion-infected mice.
  Temuulen Erdenebat; Yusuke Komatsu; Nozomi Uwamori; Misaki Tanaka; Takashi Hoshika; Takeshi Yamasaki; Ayano Shimakura; Akio Suzuki; Toyotaka Sato; Motohiro Horiuchi
  Frontiers in molecular neuroscience, 17, 1498142, 1498142, 2024, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal, The accumulation of a disease-specific isoform of prion protein (PrPSc) and histopathological lesions, such as neuronal loss, are unevenly distributed in the brains of humans and animals affected with prion diseases. This distribution varies depending on the diseases and/or the combinations of prion strain and experimental animal. The brain region-dependent distribution of PrPSc and neuropathological lesions suggests a neuronal cell-type-dependent prion propagation and vulnerability to prion infection. However, the underlying mechanism is largely unknown. In this study, we provided evidence that the prion 22L strain propagates more efficiently in excitatory neurons than inhibitory neurons and that excitatory neurons in the thalamus are vulnerable to prion infection. PrPSc accumulation was less intense in the striatum, where GABAergic inhibitory neurons predominate, compared to the cerebral cortex and thalamus, where glutamatergic excitatory neurons are predominant, in mice intracerebrally or intraperitoneally inoculated with the 22L strain. PrPSc stains were observed along the needle track after stereotaxic injection into the striatum, whereas they were also observed away from the needle track in the thalamus. Consistent with inefficient prion propagation in the striatum, the 22L prion propagated more efficiently in glutamatergic neurons than GABAergic neurons in primary neuronal cultures. RNAscope in situ hybridization revealed a decrease in Vglut1- and Vglut2-expressing neurons in the ventral posterolateral nuclei of the thalamus in 22L strain-infected mice, whereas no decrease in Vgat-expressing neurons was observed in the adjacent reticular nucleus, mainly composed of Vgat-expressing interneurons. The excitatory neuron-prone prion propagation and excitatory neuronal loss in 22L strain-infected mice shed light on the neuropathological mechanism of prion diseases.
• High prevalence of colistin heteroresistance in specific species and lineages of Enterobacter cloacae complex derived from human clinical specimens.
  Shota Fukuzawa; Toyotaka Sato; Kotaro Aoki; Soh Yamamoto; Noriko Ogasawara; Chie Nakajima; Yasuhiko Suzuki; Motohiro Horiuchi; Satoshi Takahashi; Shin-Ichi Yokota
  Annals of clinical microbiology and antimicrobials, 22, 1, 60, 60, 15 Jul. 2023, [Peer-reviewed], [International Magazine]
  English, Scientific journal, BACKGROUND: Colistin (CST) is a last-line drug for multidrug-resistant Gram-negative bacterial infections. CST-heteroresistant Enterobacter cloacae complex (ECC) has been isolated. However, integrated analysis of epidemiology and resistance mechanisms based on the complete ECC species identification has not been performed. METHODS: Clinical isolates identified as "E. cloacae complex" by MALDI-TOF MS Biotyper Compass in a university hospital in Japan were analyzed. Minimum inhibitory concentrations of CST were determined by the broth microdilution method. The population analysis profiling (PAP) was performed for detecting the heteroresistant phenotype. The heat shock protein 60 (hsp60) cluster was determined from its partial nucleotide sequence. From the data of whole-genome sequencing, average nucleotide identity (ANI) for determining ECC species, multilocus sequence type, core genome single-nucleotide-polymorphism-based phylogenetic analysis were performed. phoPQ-, eptA-, and arnT-deleted mutants were established to evaluate the mechanism underlying colistin heteroresistance. The arnT mRNA expression levels were determined by reverse transcription quantitative PCR. RESULTS: Thirty-eight CST-resistant isolates, all of which exhibited the heteroresistant phenotype by PAP, were found from 138 ECC clinical isolates (27.5%). The prevalence of CST-resistant isolates did not significantly differ among the origin of specimens (29.0%, 27.8%, and 20.2% for respiratory, urine, and blood specimens, respectively). hsp60 clusters, core genome phylogeny, and ANI revealed that the CST-heteroresistant isolates were found in all or most of Enterobacter roggenkampii (hsp60 cluster IV), Enterobacter kobei (cluster II), Enterobacter chuandaensis (clusters III and IX), and Enterobacter cloacae subspecies (clusters XI and XII). No heteroresistant isolates were found in Enterobacter hormaechei subspecies (clusters VIII, VI, and III) and Enterobacter ludwigii (cluster V). CST-induced mRNA upregulation of arnT, which encodes 4-amino-4-deoxy-L-arabinose transferase, was observed in the CST-heteroresistant isolates, and it is mediated by phoPQ pathway. Isolates possessing mcr-9 and mcr-10 (3.6% and 5.6% of total ECC isolates, respectively) exhibited similar CST susceptibility and PAP compared with mcr-negative isolates. CONCLUSIONS: Significant prevalence (approximately 28%) of CST heteroresistance is observed in ECC clinical isolates, and they are accumulated in specific species and lineages. Heteroresistance is occurred by upregulation of arnT mRNA induced by CST. Acquisition of mcr genes contributes less to CST resistance in ECC.
• Colistin-resistant bacteria poses few risks under physiological conditions
  Soh Yamamoto; Masaru Usui; Noriko Ogasawara; Wataru Hayashi; Masato Suzuki; Noriyuki Nagano; Chie Nakajima; Yasuhiko Suzuki; Motohiro Horiuchi; Satoshi Takahashi; Shin-ichi Yokota; Yutaka Tamura; Toyotaka Sato
  Research Square, Research Square Platform LLC, 21 Jun. 2023, [Peer-reviewed]
  Scientific journal, Abstract
  
  Globally, 5.0 million people die annually from infections associated with antimicrobial-resistant bacteria, most commonly Escherichia coli¹. As colistin is a last-resort antibiotic for multidrug-resistant bacterial infections, the global spread of plasmid-mediated colistin resistance genes (mcr) gene is considered a major public health risk^2-4. However, the actual health risks of colistin resistance in hazardous bacteria have never been evaluated under physiological conditions. Here, we show that the fitness/virulence and colistin resistance of the pandemic multidrug-resistant E. coli clone ST131⁵ very depending on the acquired colistin resistance determinants and differ between physiological and in vitro conditions. The fitness/virulence of ST131 was unaffected by chromosomal-gene (pmrB) mutations or the acquisition of mcr-5-harbouring plasmids in mouse models. However, the acquisition of mcr-1- or mcr-3-harbouring plasmids attenuated fitness/virulence and promoted colistin susceptibility in human serum. We identified two virulence attenuation factors (vafA and vafB) on the pIncI2_mcr-1 plasmid that hijacked the ST131 transcriptome and inhibited nucleotide synthesis, attenuating colistin resistance. Our results demonstrate that colistin resistance poses much less of a threat than believed^6,7. We suggest that “nonresistance genes,” rather than resistance genes, are important antimicrobial resistance determinants for human health because they determine fitness/virulence and ultimately antimicrobial susceptibility under physiological conditions.
• Pseudo-outbreak of Mycobacterium lentiflavum at a general hospital in Japan
  Yutaro Nagano; Koji Kuronuma; Yasuo Kitamura; Kanami Nagano; Hayato Yabe; Sayaka Kudo; Toyotaka Sato; Shinya Nirasawa; Mami Nakae; Motohiro Horiuchi; Shin-ichi Yokota; Yoshihiro Fujiya; Atsushi Saito; Satoshi Takahashi; Hirofumi Chiba
  Infection Control & Hospital Epidemiology, 44, 11, 1809, 1815, Cambridge University Press (CUP), 25 Apr. 2023, [Peer-reviewed]
  Scientific journal, Abstract
  
  Background:
  
  Mycobacterium lentiflavum is a slow-growing nontuberculous mycobacterium that is widely distributed in soil and water systems, but it is sometimes pathogenic to humans. Although cases of M. lentiflavum infections are rare, 22 isolates of M. lentiflavum were identified at a single hospital in Japan. We suspected a nosocomial outbreak; thus, we conducted transmission pattern and genotype analyses.
  
  Methods:
  
  Cases of M. lentiflavum isolated at Kushiro City General Hospital in Japan between May 2020 and April 2021 were analyzed. The patient samples and environmental culture specimens underwent whole-genome sequencing (WGS). Additionally, we retrospectively collected clinical data from patient medical records.
  
  Results:
  
  Altogether, 22 isolates of M. lentiflavum were identified from sputum and bronchoalveolar lavage samples. Clinically, the instances with M. lentiflavum isolates were considered contaminants. In the WGS analysis, 19 specimens, including 18 patient samples and 1 environmental culture from the hospital’s faucet, showed genetic similarity. The frequency of M. lentiflavum isolation decreased after we prohibited the use of taps where M. lentiflavum was isolated.
  
  Conclusions:
  
  WGS analysis identified that the cause of M. lentiflavum pseudo-outbreak was the water used for patient examinations, including bronchoscopy.
• Discovery of Biologically Optimized Polymyxin Derivatives Facilitated by Peptide Scanning and In Situ Screening Chemistry.
  Rintaro Kaguchi; Akira Katsuyama; Toyotaka Sato; Satoshi Takahashi; Motohiro Horiuchi; Shin-Ichi Yokota; Satoshi Ichikawa
  Journal of the American Chemical Society, 145, 6, 3665, 3681, 15 Feb. 2023, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Peptides can be converted to highly active compounds by introducing appropriate substituents on the suitable amino acid residue. Although modifiable residues in peptides can be systematically identified by peptide scanning methodologies, there is no practical method for optimization at the "scanned" position. With the purpose of using derivatives not only for scanning but also as a starting point for further chemical functionalization, we herein report the "scanning and direct derivatization" strategy through chemoselective acylation of embedded threonine residues by a serine/threonine ligation (STL) with the help of in situ screening chemistry. We have applied this strategy to the optimization of the polymyxin antibiotics, which were selected as a model system to highlight the power of the rapid derivatization of active scanning derivatives. Using this approach, we explored the structure-activity relationships of the polymyxins and successfully prepared derivatives with activity against polymyxin-resistant bacteria and those with Pseudomonas aeruginosa selective antibacterial activity. This strategy opens up efficient structural exploration and further optimization of peptide sequences.
• Synthesis of macrocyclic nucleoside antibacterials and their interactions with MraY.
  Takeshi Nakaya; Miyuki Yabe; Ellene H Mashalidis; Toyotaka Sato; Kazuki Yamamoto; Yuta Hikiji; Akira Katsuyama; Motoko Shinohara; Yusuke Minato; Satoshi Takahashi; Motohiro Horiuchi; Shin-Ichi Yokota; Seok-Yong Lee; Satoshi Ichikawa
  Nature communications, 13, 1, 7575, 7575, 20 Dec. 2022, [Peer-reviewed], [International Magazine]
  English, Scientific journal, The development of new antibacterial drugs with different mechanisms of action is urgently needed to address antimicrobial resistance. MraY is an essential membrane enzyme required for bacterial cell wall synthesis. Sphaerimicins are naturally occurring macrocyclic nucleoside inhibitors of MraY and are considered a promising target in antibacterial discovery. However, developing sphaerimicins as antibacterials has been challenging due to their complex macrocyclic structures. In this study, we construct their characteristic macrocyclic skeleton via two key reactions. Having then determined the structure of a sphaerimicin analogue bound to MraY, we use a structure-guided approach to design simplified sphaerimicin analogues. These analogues retain potency against MraY and exhibit potent antibacterial activity against Gram-positive bacteria, including clinically isolated drug resistant strains of S. aureus and E. faecium. Our study combines synthetic chemistry, structural biology, and microbiology to provide a platform for the development of MraY inhibitors as antibacterials against drug-resistant bacteria.
• Extended application of the rapid post-mortem test kit for bovine spongiform encephalopathy to chronic wasting disease.
  Yuichi Matsuura; Kohtaro Miyazawa; Motohiro Horiuchi; Akio Suzuki; Mayumi Yokoyama; Morikazu Imamura; Keigo Ikeda; Yoshifumi Iwamaru
  Microbiology and immunology, 66, 5, 212, 215, May 2022, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Chronic wasting disease (CWD) is a prion disease affecting cervid species primarily in the United States of America and Canada; however, it is now emerging in Scandinavian countries. Although CWD cases have not been reported in Japan, in case of a CWD outbreak occuring, it is critical to prepare for testing a large number of specimens. The present study showed that a rapid post-mortem test kit, which is used for bovine spongiform encephalopathy surveillance in Japan, is valid for the detection of CWD prion.
• Solid-Phase Total Synthesis of Plusbacin A₃
  Kazuki Takashina; Akira Katsuyama; Rintaro Kaguchi; Kazuki Yamamoto; Toyotaka Sato; Satoshi Takahashi; Motohiro Horiuchi; Shin-ichi Yokota; Satoshi Ichikawa
  Organic Letters, 24, 11, 2253, 2257, American Chemical Society (ACS), 16 Mar. 2022, [Peer-reviewed]
  Scientific journal
• Colistin Susceptibility in Companion Animal-Derived Escherichia coli, Klebsiella spp., and Enterobacter spp. in Japan: Frequent Isolation of Colistin-Resistant Enterobacter cloacae Complex.
  Toyotaka Sato; Kazuki Harada; Masaru Usui; Shin-Ichi Yokota; Motohiro Horiuchi
  Frontiers in cellular and infection microbiology, 12, 946841, 946841, 2022, [International Magazine]
  English, Scientific journal, Transmission of colistin-resistant Enterobacterales from companion animals to humans poses a clinical risk as colistin is a last-line antimicrobial agent for treatment of multidrug-resistant Gram-negative bacteria including Enterobacterales. In this study, we investigated the colistin susceptibility of 285 Enterobacterales (including 140 Escherichia coli, 86 Klebsiella spp., and 59 Enterobacter spp.) isolated from companion animals in Japan. We further characterized colistin-resistant isolates by multilocus sequence typing (MLST), phylogenetic analysis of hsp60 sequences, and population analysis profiling, to evaluate the potential clinical risk of companion animal-derived colistin-resistant Enterobacterales to humans in line with the One Health approach. All E. coli isolates were susceptible to colistin, and only one Klebsiella spp. isolate (1.2%, 1/86 isolates) was colistin resistant. Enterobacter spp. isolates were frequently colistin resistant (20.3%, 12/59 isolates). In colistin-resistant Enterobacter spp., all except one isolate exhibited colistin heteroresistance by population analysis profiling. These colistin-heteroresistant isolates belonged to clusters I, II, IV, VIII, and XII based on hsp60 phylogeny. MLST analysis revealed that 12 colistin-resistant Enterobacter spp. belonged to the Enterobacter cloacae complex; five Enterobacter kobei (four ST591 and one ST1577), three Enterobacter asburiae (one ST562 and two ST1578), two Enterobacter roggenkampii (ST606 and ST1576), and Enterobacter hormaechei (ST1579) and E. cloacae (ST765) (each one strain). Forty-two percent of the colistin-resistant E. cloacae complex isolates (predominantly ST562 and ST591) belonged to lineages with human clinical isolates. Four E. kobei ST591 isolates were resistant to third-generation cephalosporines, aminoglycosides, and fluroquinolones but remained susceptible to carbapenems. In conclusion, our study is the first to our knowledge to report the frequent isolation of the colistin-resistant E. cloacae complex from companion animals. Furthermore, a subset of isolates belonged to human-associated lineages with resistance to multiple classes of antibiotics. These data warrant monitoring carriage of the colistin-resistant E. cloacae complex in companion animals as part of a domestic infection control procedure in line with the One Health approach.
• Isolation of Human Lineage, Fluoroquinolone-Resistant and Extended-β-Lactamase-Producing Escherichia coli Isolates from Companion Animals in Japan
  Toyotaka Sato; Shin-ichi Yokota; Tooru Tachibana; Satoshi Tamai; Shigeki Maetani; Yutaka Tamura; Motohiro Horiuchi
  Antibiotics, 10, 12, 1463, 1463, MDPI AG, 28 Nov. 2021, [Peer-reviewed]
  Scientific journal, An increase in human and veterinary fluoroquinolone-resistant Escherichia coli is a global concern. In this study, we isolated fluoroquinolone-resistant E. coli isolates from companion animals and characterized them using molecular epidemiological analysis, multiplex polymerase chain reaction to detect E. coli ST131 and CTX-M type extended-spectrum β-lactamases (ESBL), and multi-locus sequence typing analysis. Using plain-CHROMagar ECC, 101 E. coli isolates were isolated from 34 rectal swabs of dogs and cats. The prevalence of resistance to fluoroquinolone and cefotaxime was 27.7% and 24.8%, respectively. The prevalence of fluoroquinolone-resistant isolates (89.3%) was higher when CHROMagar ECC with CHROMagar ESBL supplement was used for E. coli isolation. The prevalence of cefotaxime resistance was also higher (76.1%) when 1 mg/L of ciprofloxacin-containing CHROMagar ECC was used for isolation. The cefotaxime-resistant isolates possessed CTX-M type β-lactamase genes (CTX-M-14, CTX-M-15, or CTX-M-27). Seventy-five percent of fluoroquinolone-resistant isolates were sequence types ST131, ST10, ST1193, ST38, or ST648, which are associated with extensive spread in human clinical settings. In addition, we isolated three common fluoroquinolone-resistant E. coli lineages (ST131 clade C1-M-27, C1-nM27 and ST2380) from dogs and their respective owners. These observations suggest that companion animals can harbor fluoroquinolone-resistant and/or ESBL-producing E. coli, in their rectums, and that transmission of these isolates to their owners can occur.
• Refeeding activates neurons in the dorsomedial hypothalamus to inhibit food intake and promote positive valence.
  Daigo Imoto; Izumi Yamamoto; Hirokazu Matsunaga; Toya Yonekura; Ming-Liang Lee; Kan X Kato; Takeshi Yamasaki; Shucheng Xu; Taiga Ishimoto; Satoshi Yamagata; Ken-Ichi Otsuguro; Motohiro Horiuchi; Norifumi Iijima; Kazuhiro Kimura; Chitoku Toda
  Molecular metabolism, 54, 101366, 101366, 30 Oct. 2021, [Peer-reviewed], [International Magazine]
  English, Scientific journal, OBJECTIVE: The regulation of food intake is a major research area in the study of obesity, which plays a key role in the development of metabolic syndrome. Gene targeting studies have clarified the roles of hypothalamic neurons in feeding behavior, but the deletion of a gene has a long-term effect on neurophysiology. Our understanding of short-term changes such as appetite under physiological conditions is therefore still limited. METHODS: Targeted recombination in active populations (TRAP) is a newly developed method for labeling active neurons by using tamoxifen-inducible Cre recombination controlled by the promoter of activity-regulated cytoskeleton-associated protein (Arc/Arg3.1), a member of immediate early genes. Transgenic mice for TRAP were fasted overnight, re-fed with normal diet, and injected with 4-hydroxytamoxifen 1 h after the refeeding to label the active neurons. The role of labeled neurons was examined by expressing excitatory or inhibitory designer receptors exclusively activated by designer drugs (DREADDs). The labeled neurons were extracted and RNA sequencing was performed to identify genes that are specifically expressed in these neurons. RESULTS: Fasting-refeeding activated and labeled neurons in the compact part of the dorsomedial hypothalamus (DMH) that project to the paraventricular hypothalamic nucleus. Chemogenetic activation of the labeled DMH neurons decreased food intake and developed place preference, an indicator of positive valence. Chemogenetic activation or inhibition of these neurons had no influence on the whole-body glucose metabolism. The labeled DMH neurons expressed prodynorphin (pdyn), gastrin-releasing peptide (GRP), cholecystokinin (CCK), and thyrotropin-releasing hormone receptor (Trhr) genes. CONCLUSIONS: We identified a novel cell type of DMH neurons that can inhibit food intake and promote feeding-induced positive valence. Our study provides insight into the role of DMH and its molecular mechanism in the regulation of appetite and emotion.
• Monitoring of chronic wasting disease using real-time quaking-induced conversion assay in Japan.
  Akio Suzuki; Kazuhei Sawada; Temuulen Erdenebat; Takeshi Yamasaki; Minoru Tobiume; Kinuyo Suga; Motohiro Horiuchi
  The Journal of veterinary medical science, 24 Sep. 2021, [Peer-reviewed], [Corresponding author], [Domestic magazines]
  English, Scientific journal, There has been no report on Chronic wasting disease (CWD) cases in Japan to date; however, there is concern about the geographic spread of CWD. To clarify the CWD status in Japan, we conducted CWD monitoring using real-time quaking-induced conversion (RT-QuIC) assay which can detect the low level of CWD prions. A total of 690 obex samples collected from sika deer and Reeves's muntjac in Hokkaido and Honshu was tested for CWD prions. No CWD-positive cases were found, suggesting that CWD is nonexistent in Japan. Our results also indicate that RT-QuIC assay is useful for continuous monitoring of CWD. Furthermore, nucleotide sequence analysis of the PrP gene revealed sika deer in Japan harbor CWD susceptible allele.
• Research Note: Antimicrobial resistance of Campylobacter species isolated from chickens near Ulaanbaatar city, Mongolia.
  Zolzaya Byambajav; Erdenebat Bulgan; Yuji Hirai; Momoko Nakayama; Misaki Tanaka; Yurika Nitta; Akio Suzuki; Takashi Umemura; Bold Altankhuu; Alimaa Tsagaan; Batbaatar Vanaabaatar; Erdenebaatar Janchivdorj; Nyam-Osor Purevdorj; Narantuya Ayushjav; Takeshi Yamasaki; Motohiro Horiuchi
  Poultry science, 100, 3, 100916, 100916, Mar. 2021, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal, There has been no report on the prevalence of Campylobacter spp. in farm animals in Mongolia. To uncover the prevalence of Campylobacter spp. in chickens in Mongolia and their antimicrobial resistance, in this study, we isolated and characterized Campylobacter spp. from chickens in Mongolia. We collected 71 cloacal swabs of chickens from 5 farms including 4 layer farms and one broiler farm near Ulaanbaatar city and isolated 25 Campylobacter jejuni and 6 Campylobacter coli isolates. All isolates were resistant to tetracycline, and 3 C. coli isolates were resistant to erythromycin. The C. coli isolates possessed either the erm(B) gene or nucleotide substitution at nt 2,075 of 23S rDNA, both of which are known to be associated with erythromycin resistance. Sixteen of the 31 C. jejuni/C. coli isolates (51.6%) were resistant to nalidixic acid and fluoroquinolones. All the fluoroquinolone-resistant isolates possessed amino acid substitution from threonine to isoleucine at codon 86 (nucleotide substitution: ACA to ATA). Multilocus sequence typing and phylogenetic analyses showed a variation in C. jejuni/C. coli in chickens in Mongolia. In addition, some of the C. jejuni isolates seemed to be phylogenetically close to isolates in Asian and Oceanian countries. This is the first report on the characterization of antimicrobial resistance of Campylobacter spp. in farm animals in Mongolia and is valuable for implementation of measures for a prudent use of antimicrobials in farm animals.
• Relationship between lactational performance and metabolic parameters of Mongolian native grazing mares
  Badrakh SANDAGDORJ; Tserenpil BAIGALMAA; Sedhuu BURENJARGAL; Motohiro HORIUCHI; Munkhbat ENKHDALAI; Davaakhuu BAYANBAT; Dashdorj JANCHIV; Ooyo JAMYANDORJ; Purevdorj ULZII-ORSHIKH; Purevdorj NYAM-OSOR
  Journal of Equine Science, 32, 3, 91, 98, Japan Society of Equine Science, 2021, [Peer-reviewed]
  Scientific journal
• Involvement of N- and C-terminal region of recombinant cervid prion protein in its reactivity to CWD and atypical BSE prions in real-time quaking-induced conversion reaction in the presence of high concentrations of tissue homogenates.
  Akio Suzuki; Kazuhei Sawada; Takeshi Yamasaki; Nathaniel D Denkers; Candace K Mathiason; Edward A Hoover; Motohiro Horiuchi
  Prion, 14, 1, 283, 295, Dec. 2020, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal, The real-time quaking-induced conversion (RT-QuIC) reaction is a sensitive and specific method for detecting prions. However, inhibitory factors present in tissue homogenates can easily interfere with this reaction. To identify the RT-QuIC condition under which low levels of chronic wasting disease (CWD) and bovine spongiform encephalopathy (BSE) prions can be detected in the presence of high concentrations of brain tissue homogenates, reactivities of various recombinant prion proteins (rPrPs) were tested. Among the tested rPrPs, recombinant cervid PrP (rCerPrP) showed a unique reactivity: the reactivity of rCerPrP to CWD and atypical BSE prions was not highly affected by high concentrations of normal brain homogenates. The unique reactivity of rCerPrP disappeared when the N-terminal region (aa 25-93) was truncated. Replacement of aa 23-149 of mouse (Mo) PrP with the corresponding region of CerPrP partially restored the unique reactivity of rCerPrP in RT-QuIC. Replacement of the extreme C-terminal region of MoPrP aa 219-231 to the corresponding region of CerPrP partially conferred the unique reactivity of rCerPrP to rMoPrP, suggesting the involvement of both N- and C-terminal regions. Additionally, rCerN-Mo-CerCPrP, a chimeric PrP comprising CerPrP aa 25-153, MoPrP aa 150-218, and CerPrP aa 223-233, showed an additive effect of the N- and C-terminal regions. These results provide a mechanistic implication for detecting CWD and atypical BSE prions using rCerPrP and are useful for further improvements of RT-QuIC.
• Selective neuronal vulnerability is involved in cerebellar lesions of Guinea pigs infected with bovine spongiform encephalopathy (BSE) prions: Immunohistochemical and electron microscopic investigations.
  Shoichi Sakaguchi; Sayo Shintani; Kyohei Kamio; Akio Sekiya; Satomi Kato; Yoshikage Muroi; Motohiro Horiuchi; Hidefumi Furuoka
  Neuropathology : official journal of the Japanese Society of Neuropathology, 40, 2, 167, 179, Apr. 2020, [Peer-reviewed], [International Magazine]
  English, Scientific journal, The cerebellar lesions of bovine spongiform encephalopathy (BSE)-infected guinea pigs were characterized as severe atrophy of the cerebellar cortex associated with the loss of granule cells, decrease in the width of the molecular layer, and intense protease-resistant prion protein (PrPSc ) accumulations that are similar to cerebellar lesions in kuru and the VV2 type of sporadic Creutzfeldt-Jakob disease. The aim of this study is to assess the relationships between the distribution and localization of PrPSc and synapses expressing neurotransmitter transporters in order to reveal the pathogenesis of the disease. We used cell-type-specific immunohistochemical makers recognizing glutamatergic and γ-aminobutylic acid (GABA)ergic terminals to identify terminals impaired with PrPSc accumulations. The distribution of PrPSc accumulations and immunoreactivity of synaptic vesicles were studied throughout the neuroanatomical pathways in cerebellar lesions. Time course study demonstrated that PrPSc accumulation showed a tendency to spread from granular layer to molecular layer. The immunoreactivity of vesicular glutamate transporter 1 (VGluT1) was localized in axon terminals of cerebellar granule cells, and decreased in association with the severity of PrPSc accumulations and loss of granule cells. Immunoreactivities of vesicular glutamate transporter 2 (VGluT2) and vesicular GABA transporter (VGAT) that exist in axon terminals of inferior olivary neurons and GABAergic synapses of Purkinje cells, respectively, were preserved well in these lesions. In brainstem, VGluT1 immunoreactivity decreased selectively in pontine nuclei that are a component of the pontocerebellar pathway, although other neurotransmitter immunoreactivities were preserved well. Our findings suggest that the selective loss of VGluT1-immunoreactive synapses subsequent to PrPSc accumulations can contribute to the pathogenesis of cerebellar lesions of BSE-infected guinea pigs.
• Enhanced phosphorylation of PERK in primary cultured neurons as an autonomous neuronal response to prion infection.
  Misaki Tanaka; Takeshi Yamasaki; Rie Hasebe; Akio Suzuki; Motohiro Horiuchi
  PloS one, 15, 6, e0234147, 2020, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal, Conversion of cellular prion protein (PrPC) into the pathogenic isoform of prion protein (PrPSc) in neurons is one of the key pathophysiological events in prion diseases. However, the molecular mechanism of neurodegeneration in prion diseases has yet to be fully elucidated because of a lack of suitable experimental models for analyzing neuron-autonomous responses to prion infection. In the present study, we used neuron-enriched primary cultures of cortical and thalamic mouse neurons to analyze autonomous neuronal responses to prion infection. PrPSc levels in neurons increased over the time after prion infection; however, no obvious neuronal losses or neurite alterations were observed. Interestingly, a finer analysis of individual neurons co-stained with PrPSc and phosphorylated protein kinase RNA-activated-like endoplasmic reticulum (ER) kinase (p-PERK), the early cellular response of the PERK-eukaryotic initiation factor 2 (eIF2α) pathway, demonstrated a positive correlation between the number of PrPSc granular stains and p-PERK granular stains, in cortical neurons at 21 dpi. Although the phosphorylation of PERK was enhanced in prion-infected cortical neurons, there was no sign of subsequent translational repression of synaptic protein synthesis or activations of downstream unfolded protein response (UPR) in the PERK-eIF2α pathway. These results suggest that PrPSc production in neurons induces ER stress in a neuron-autonomous manner; however, it does not fully activate UPR in prion-infected neurons. Our findings provide insights into the autonomous neuronal responses to prion propagation and the involvement of neuron-non-autonomous factor(s) in the mechanisms of neurodegeneration in prion diseases.
• Bergeyella zoohelcum isolated from oral cavities of therapy dogs.
  Yasukazu Muramatsu; Nami Haraya; Kazuki Horie; Leo Uchida; Takanori Kooriyama; Akio Suzuki; Motohiro Horiuchi
  Zoonoses and public health, 66, 8, 936, 942, Dec. 2019, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Bergeyella zoohelcum causes rare but severe human clinical diseases, which mostly arise from animal bites. Notably, Bergeyella infections can also occur in older people after prolonged exposure to dogs or cats without biting. We detected B. zoohelcum in oral cavities of therapy dogs in close contact with older people residing in nursing homes. Twenty-two bacterial isolates were identified as B. zoohelcum by using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) and 16S rRNA gene sequencing. Our results showed that MALDI-TOF MS is an effective tool for rapid identification of rarely isolated, difficult-to-identify microorganisms, such as B. zoohelcum, derived from not only human clinical samples but also animal samples. To our knowledge, this is the first report on detection of B. zoohelcum from therapy dogs. We have provided information on dog-assisted therapy to improve the relationship between humans and animals in ageing societies, particularly for preventive healthcare of older people living in nursing care facilities.
• The first isolation and identification of canine parvovirus (CPV) type 2c variants during 2016-2018 genetic surveillance of dogs in Mongolia
  Uyangaa Temuujin; Ariunaa Tserendorj; Jumpei Fujiki; Yoshihiro Sakoda; Erdene-Ochir Tseren-Ochir; Masatoshi Okamatsu; Keita Matsuno; Tumenjargal Sharav; Motohiro Horiuchi; Takashi Umemura; Tungalag Chultemdorj
  INFECTION GENETICS AND EVOLUTION, 73, 269, 275, Sep. 2019, [Peer-reviewed], [International Magazine]
  English, Scientific journal
• Estimation of prion infectivity in tissues of cattle infected with atypical BSE by real time-quaking induced conversion assay.
  Kazuhei Sawada; Akio Suzuki; Takeshi Yamasaki; Yoshifumi Iwamaru; Yuichi Matsuura; Kohtaro Miyazawa; Kentaro Masujin; Ryuichiro Atarashi; Motohiro Horiuchi
  The Journal of veterinary medical science, 81, 6, 846, 850, 06 Jun. 2019, [Peer-reviewed], [Corresponding author], [Domestic magazines]
  English, Scientific journal, Atypical bovine spongiform encephalopathy (BSE), first identified in 2004, poses a threat due to the potential to spread the disease to cattle and other animals, including humans. Here, we estimated prion titers in various tissues of cattle infected with atypical BSE using a real-time quaking-induced conversion assay that detects amyloid seeding activity of a disease-specific prion protein, PrPSc, a major component of prions. PrPSc was detected both in and outside of nerve tissues, and some of the peripheral nerve tissues contained relatively high prion titers. Low titers of prions were also observed in masseter, jejunum, and adrenal glands. Quantitative data on prion infectivity in tissues of atypical BSE-affected cattle is useful to assess the risk of atypical BSE.
• Enhancement of binding avidity by bivalent binding enables PrPSc-specific detection by anti-PrP monoclonal antibody 132.
  Akio Suzuki; Takeshi Yamasaki; Rie Hasebe; Motohiro Horiuchi
  PloS one, 14, 6, e0217944, 2019, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal
• Tracking and clarifying differential traits of classical- and atypical L-type bovine spongiform encephalopathy prions after transmission from cattle to cynomolgus monkeys.
  Ken'ichi Hagiwara; Yuko Sato; Yoshio Yamakawa; Hideyuki Hara; Minoru Tobiume; Yuko Okemoto-Nakamura; Tetsutaro Sata; Motohiro Horiuchi; Hiroaki Shibata; Fumiko Ono
  PloS one, 14, 5, e0216807, 2019, [Peer-reviewed], [International Magazine]
  English, Scientific journal, Classical- (C-) and atypical L-type bovine spongiform encephalopathy (BSE) prions cause different pathological phenotypes in cattle brains, and the disease-associated forms of each prion protein (PrPSc) has a dissimilar biochemical signature. Bovine C-BSE prions are the causative agent of variant Creutzfeldt-Jakob disease. To date, human infection with L-BSE prions has not been reported, but they can be transmitted experimentally from cows to cynomolgus monkeys (Macaca fascicularis), a non-human primate model. When transmitted to monkeys, C- and L-BSE prions induce different pathological phenotypes in the brain. However, when isolated from infected brains, the two prion proteins (PrPSc) have similar biochemical signatures (i.e., electrophoretic mobility, glycoforms, and resistance to proteinase K). Such similarities suggest the possibility that L-BSE prions alter their virulence to that of C-BSE prions during propagation in monkeys. To clarify this possibility, we conducted bioassays using inbred mice. C-BSE prions with or without propagation in monkeys were pathogenic to mice, and exhibited comparable incubation periods in secondary passage in mice. By contrast, L-BSE prions, either with or without propagation in monkeys, did not cause the disease in mice, indicating that the pathogenicity of L-BSE prions does not converge towards a C-BSE prion type in this primate model. These results suggest that, although C- and L-BSE prions propagated in cynomolgus monkeys exhibit similar biochemical PrPSc signatures and consist of the monkey amino acid sequence, the two prions maintain strain-specific conformations of PrPSc in which they encipher and retain unique pathogenic traits.
• Retrograde Transport by Clathrin-Coated Vesicles is Involved in Intracellular Transport of PrPSc in Persistently Prion-Infected Cells.
  Takeshi Yamasaki; Akio Suzuki; Rie Hasebe; Motohiro Horiuchi
  Scientific reports, 8, 1, 12241, 12241, 16 Aug. 2018, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal
• Flow Cytometric Detection of PrPSc in Neurons and Glial Cells from Prion-Infected Mouse Brains.
  Takeshi Yamasaki; Akio Suzuki; Rie Hasebe; Motohiro Horiuchi
  Journal of virology, 92, 1, 01 Jan. 2018, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal
• Therapeutic effect of autologous compact bone-derived mesenchymal stem cell transplantation on prion disease.
  Zhifu Shan; Yuji Hirai; Momoko Nakayama; Ryo Hayashi; Takeshi Yamasaki; Rie Hasebe; Chang-Hyun Song; Motohiro Horiuchi
  The Journal of general virology, 98, 10, 2615, 2627, Oct. 2017, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal
• Listeria monocytogenes serotype 4b strains replicate in monocytes/macrophages more than the other serotypes
  Rie Hasebe; Ryo Nakao; Aiko Ohnuma; Takeshi Yamasaki; Hirofumi Sawa; Shinji Takai; Motohiro Horiuchi
  JOURNAL OF VETERINARY MEDICAL SCIENCE, 79, 6, 962, 969, Jun. 2017, [Peer-reviewed]
  English, Scientific journal
• Complement factors alter the amount of PrP(Sc) in primary-cultured mouse cortical neurons associated with increased membrane permeability.
  Rie Hasebe; Misaki Tanaka; Akio Suzuki; Takeshi Yamasaki; Motohiro Horiuchi
  Virology, 496, 9, 20, Sep. 2016, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal
• Comparison of abnormal isoform of prion protein in prion-infected cell lines and primary-cultured neurons by PrPSc-specific immunostaining.
  Misaki Tanaka; Ai Fujiwara; Akio Suzuki; Takeshi Yamasaki; Rie Hasebe; Kentaro Masujin; Motohiro Horiuchi
  The Journal of general virology, 97, 8, 2030, 2042, Aug. 2016, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal
• Establishment of a simple cell-based ELISA for the direct detection of abnormal isoform of prion protein from prion-infected cells without cell lysis and proteinase K treatment.
  Zhifu Shan; Takeshi Yamasaki; Akio Suzuki; Rie Hasebe; Motohiro Horiuchi
  Prion, 10, 4, 305, 18, 03 Jul. 2016, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal
• PrPSc-Specific Antibody Reveals C-Terminal Conformational Differences between Prion Strains.
  Eri Saijo; Andrew G Hughson; Gregory J Raymond; Akio Suzuki; Motohiro Horiuchi; Byron Caughey
  Journal of virology, 90, 10, 4905, 4913, 15 May 2016, [Peer-reviewed], [International Magazine]
  English, Scientific journal
• Prion Protein Binds to Aldolase A Produced by Bovine Intestinal M Cells
  Yuya Nagasawa; Yu Takahashi; Wataru Itani; Hitoshi Watanabe; Yusuke Hidaka; Shotaro Morita; Kei Suzuki; Kouichi Watanabe; Shyuichi Ohwada; Haruki Kitazawa; Morikazu Imamura; Takashi Yokoyama; Motohiro Horiuchi; Suehiro Sakaguchi; Shirou Mohri; Michael T. Rose; Tomonori Nochi; Hisashi Aso
  Open Journal of Veterinary Medicine, 05, 03, 43, 60, Scientific Research Publishing, Inc., 2015, [Peer-reviewed]
  Scientific journal
• Temporary upregulation of anti-inflammatory cytokine IL-13 expression in the brains of CD14 deficient mice in the early stage of prion infection.
  Rie Hasebe; Akio Suzuki; Takeshi Yamasaki; Motohiro Horiuchi
  Biochemical and biophysical research communications, 454, 1, 125, 30, 07 Nov. 2014, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal
• Genetic diversity in the prion protein gene (PRNP) of domestic cattle and water buffaloes in Vietnam, Indonesia and Thailand.
  Leo Uchida; Agus Heriyanto; Chalermchaikit Thongchai; Tran Thi Hanh; Motohiro Horiuchi; Kanako Ishihara; Yutaka Tamura; Yasukazu Muramatsu
  The Journal of veterinary medical science, 76, 7, 1001, 8, Jul. 2014, [Peer-reviewed], [Domestic magazines]
  English, Scientific journal
• Characterization of intracellular dynamics of inoculated PrP-res and newly generated PrP(Sc) during early stage prion infection in Neuro2a cells.
  Takeshi Yamasaki; Gerald S Baron; Akio Suzuki; Rie Hasebe; Motohiro Horiuchi
  Virology, 450-451, 324, 35, Feb. 2014, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal
• Comparison of the anti-prion mechanism of four different anti-prion compounds, anti-PrP monoclonal antibody 44B1, pentosan polysulfate, chlorpromazine, and U18666A, in prion-infected mouse neuroblastoma cells.
  Takeshi Yamasaki; Akio Suzuki; Rie Hasebe; Motohiro Horiuchi
  PloS one, 9, 9, e106516, 2014, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal
• Absence of CD14 delays progression of prion diseases accompanied by increased microglial activation.
  Keiko Sakai; Rie Hasebe; Yusuke Takahashi; Chang-Hyun Song; Akio Suzuki; Takeshi Yamasaki; Motohiro Horiuchi
  Journal of virology, 87, 24, 13433, 45, Dec. 2013, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal
• Therapeutic effect of peripheral administration of an anti-prion protein antibody on mice infected with prions.
  Natsuo Ohsawa; Chang-Hyun Song; Akio Suzuki; Hidefumi Furuoka; Rie Hasebe; Motohiro Horiuchi
  Microbiology and immunology, 57, 4, 288, 97, Apr. 2013, [Peer-reviewed], [Corresponding author], [International Magazine]
  English, Scientific journal
• Cutting Edge: Brucella abortus exploits a cellular prion protein on intestinal M cells as an invasive receptor.
  Gaku Nakato; Koji Hase; Michio Suzuki; Masanobu Kimura; Manabu Ato; Misaho Hanazato; Minoru Tobiume; Motohiro Horiuchi; Ryuichiro Atarashi; Noriyuki Nishida; Masahisa Watarai; Koichi Imaoka; Hiroshi Ohno
  Journal of immunology (Baltimore, Md. : 1950), 189, 4, 1540, 4, 15 Aug. 2012, [Peer-reviewed], [International Magazine]
  English, Scientific journal
• Alternative BSE Risk Assessment Methodology for Beef and Beef Offal Imported into Japan
  Yasuhiro Yoshikawa; Motohiro Horiuchi; Naotaka Ishiguro; Mutsuyo Kadohira; Satoshi Kai; Hidehiro Mizusawa; Chisato Nagata; Takashi Onodera; Tetsutaro Sata; Toshiyuki Tsutsui; Masahito Yamada; Shigeki Yamamoto
  JOURNAL OF VETERINARY MEDICAL SCIENCE, 74, 8, 959, 968, Aug. 2012, [Peer-reviewed]
  English
• Characterization of intracellular localization of PrPSc in prion-infected cells using a mAb that recognizes the region consisting of aa 119-127 of mouse PrP
  Takeshi Yamasaki; Akio Suzuki; Takeshi Shimizu; Masahisa Watarai; Rie Hasebe; Motohiro Horiuchi
  JOURNAL OF GENERAL VIROLOGY, 93, 3, 668, 680, Mar. 2012, [Peer-reviewed], [Corresponding author]
  English, Scientific journal
• Reaction of complement factors varies with prion strains in vitro and in vivo
  Rie Hasebe; Gregory J. Raymond; Motohiro Horiuchi; Byron Caughey
  VIROLOGY, 423, 2, 205, 213, Feb. 2012, [Peer-reviewed]
  English, Scientific journal
• Identification of Chemoattractive Factors Involved in the Migration of Bone Marrow-Derived Mesenchymal Stem Cells to Brain Lesions Caused by Prions
  Chang-Hyun Song; Osamu Honmou; Hidefumi Furuoka; Motohiro Horiuchi
  JOURNAL OF VIROLOGY, 85, 21, 11069, 11078, Nov. 2011, [Peer-reviewed], [Corresponding author]
  English, Scientific journal
• Predominant Involvement of the Cerebellum in Guinea Pigs Infected with Bovine Spongiform Encephalopathy (BSE)
  H. Furuoka; M. Horiuchi; Y. Yamakawa; T. Sata
  JOURNAL OF COMPARATIVE PATHOLOGY, 144, 4, 269, 276, May 2011, [Peer-reviewed]
  English, Scientific journal
• Successive Deaths of a Captive Snow Leopard (Uncia uncia) and a Serval (Leptailurus serval) by Infection with Feline Panleukopenia Virus at Sapporo Maruyama Zoo
  Yukiko Sassa; Hideaki Yamamoto; Masami Mochizuki; Takashi Umemura; Motohiro Horiuchi; Naotaka Ishiguro; Takayuki Miyazawa
  JOURNAL OF VETERINARY MEDICAL SCIENCE, 73, 4, 491, 494, Apr. 2011, [Peer-reviewed], [Domestic magazines]
  English, Scientific journal
• The effects of lysosomal and proteasomal inhibitors on abnormal forms of prion protein degradation in murine macrophages
  Yukiko Sassa; Takeshi Yamasaki; Motohiro Horiuchi; Yasuo Inoshima; Naotaka Ishiguro
  MICROBIOLOGY AND IMMUNOLOGY, 54, 12, 763, 768, Dec. 2010, [Peer-reviewed]
  English, Scientific journal
• Transcellular transport of West Nile virus-like particles across human endothelial cells depends on residues 156 and 159 of envelope protein
  Rie Hasebe; Tadaki Suzuki; Yoshinori Makino; Manabu Igarashi; Satoko Yamanouchi; Akihiko Maeda; Motohiro Horiuchi; Hirofumi Sawa; Takashi Kimura
  BMC MICROBIOLOGY, 10, 165, Jun. 2010, [Peer-reviewed]
  English, Scientific journal
• ImmunoAT method: An initial assessment for the detection of abnormal isoforms of prion protein in formalin-fixed and paraffin-embedded tissues
  Yuko Sato; Nozomi Shimonohara; Ken-Ichi Hanaki; Motoki Goto; Yoshio Yamakawa; Motohiro Horiuchi; Hidehiro Takahashi; Tetsutaro Sata; Noriko Nakajima
  JOURNAL OF VIROLOGICAL METHODS, 165, 2, 261, 267, May 2010, [Peer-reviewed]
  English, Scientific journal
• A novel copper(II) coordination at His186 in full-length murine prion protein
  Yasuko Watanabe; Wakako Hiraoka; Manabu Igarashi; Kimihito Ito; Yuhei Shimoyama; Motohiro Horiuchi; Tohru Yamamoria; Hironobu Yasui; Mikinori Kuwabara; Fuyuhiko Inagaki; Osamu Inanami
  BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 394, 3, 522, 528, Apr. 2010, [Peer-reviewed]
  English, Scientific journal
• Cell density-dependent increase in the level of protease-resistant prion protein in prion-infected Neuro2a mouse neuroblastoma cells
  Satoshi Nakamitsu; Aya Kurokawa; Takeshi Yamasaki; Masahide Uryu; Rie Hasebe; Motohiro Horiuchi
  JOURNAL OF GENERAL VIROLOGY, 91, 2, 563, 569, Feb. 2010, [Peer-reviewed], [Corresponding author]
  English, Scientific journal
• Confomational Analysis of Soluble Oligomers of GFP Tagged Prion Protein By Fluorescence Fluctuation Spectroscopy
  Hiroshi Sakata; Motohiro Horiuchi; Izumi Takahashi; Masataka Kinjo
  CURRENT PHARMACEUTICAL BIOTECHNOLOGY, 11, 1, 87, 95, Jan. 2010, [Peer-reviewed]
  English
• Generation of monoclonal antibody that distinguishes PrPSc from PrPC and neutralizes prion infectivity
  Motohiro Horiuchi; Ayako Karino; Hidefumi Furuoka; Naotaka Ishiguro; Kumiko Kimura; Morikazu Shinagawa
  VIROLOGY, 394, 2, 200, 207, Nov. 2009, [Peer-reviewed], [Corresponding author]
  English, Scientific journal
• Effect of Transplantation of Bone Marrow-Derived Mesenchymal Stem Cells on Mice Infected with Prions
  Chang-Hyun Song; Osamu Honmou; Natsuo Ohsawa; Kiminori Nakamura; Hirofumi Hamada; Hidefumi Furuoka; Rie Hasebe; Motohiro Horiuchi
  JOURNAL OF VIROLOGY, 83, 11, 5918, 5927, Jun. 2009, [Peer-reviewed], [Corresponding author]
  English, Scientific journal
• The Region Approximately between Amino Acids 81 and 137 of Proteinase K-Resistant PrPSc Is Critical for the Infectivity of the Chandler Prion Strain
  Ryo Shindoh; Chan-Lan Kim; Chang-Hyun Song; Rie Hasebe; Motohiro Horiuchi
  JOURNAL OF VIROLOGY, 83, 8, 3852, 3860, Apr. 2009, [Peer-reviewed], [Corresponding author]
  English, Scientific journal
• 1SA2-02 Intracellular localization of abnormal isoform of prion protein(1SA2 BSJ&ABA Joint Symposium, "Prion and Virus Infections",The 47th Annual Meeting of the Biophysical Society of Japan)
  Horiuchi Motohiro
  Seibutsu Butsuri, 49, supplement, S3, The Biophysical Society of Japan General Incorporated Association, 2009
  English
• Heat shock cognate protein 70 contributes to Brucella invasion into trophoblast giant cells that cause infectious abortion
  Kenta Watanabe; Masato Tachibana; Satoshi Tanaka; Hidefumi Furuoka; Motohiro Horiuchi; Hiroshi Suzuki; Masahisa Watarai
  BMC MICROBIOLOGY, 8, 212, Dec. 2008, [Peer-reviewed]
  English, Scientific journal
• Evaluation of Methods for Removing Central Nervous System Tissue Contamination from the Surface of Beef Carcasses after Splitting
  Naoko Takada; Motohiro Horiuchi; Tetslitaro Sata; Yasushi Sawada
  JOURNAL OF VETERINARY MEDICAL SCIENCE, 70, 11, 1225, 1230, Nov. 2008, [Peer-reviewed]
  English, Scientific journal
• Effect of intraventricular infusion of anti-prion protein monoclonal antibodies on disease progression in prion-infected mice
  Chang-Hyun Song; Hidefumi Furuoka; Chan-Lan Kim; Michiko Ogino; Akio Suzuki; Rie Hasebe; Motohiro Horiuchi
  JOURNAL OF GENERAL VIROLOGY, 89, 6, 1533, 1544, Jun. 2008, [Peer-reviewed], [Corresponding author]
  English, Scientific journal
• Frequencies of PRNP Gene Polymorphisms in Vietnamese Dairy Cattle for Potential Association with BSE
  Y. Muramatsu; Y. Sakemi; M. Horiuchi; T. Ogawa; K. Suzuki; M. Kanameda; T. T. Hanh; Y. Tamura
  Zoonoses and Public Health, 55, 5, 267, 273, Wiley, 09 Apr. 2008, [Peer-reviewed]
  English, Scientific journal
• Instability of familial spongiform encephalopathy-related prion mutants
  Yasuko Watanabe; Wakako Hiraoka; Yuhei Shimoyama; Motohiro Horiuchi; Mikinori Kuwabara; Osamu Inanami
  BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 366, 1, 244, 249, Feb. 2008, [Peer-reviewed]
  English, Scientific journal
• Frequencies of PrP genotypes in meat breeds of Japanese sheep and trail of selective breeding in experimental sheep flock
  Jiro Ohara; Tetsuro Togari; Aya Kurokawa; Junko Maeda; Naotaka Ishiguro; Hidefumi Furuoka; Motohiro Horiuchi
  JOURNAL OF VETERINARY MEDICAL SCIENCE, 69, 12, 1325, 1329, Dec. 2007, [Peer-reviewed], [Corresponding author]
  English, Scientific journal
• Detection of prion protein immune complex for bovine spongiform encephalopathy diagnosis using fluorescence correlation spectroscopy and fluorescence cross-correlation spectroscopy
  Fumihiko Fujii; Motohiro Horiuchi; Masayoshi Ueno; Hiroshi Sakata; Issel Nagao; Mamoru Tamura; Masataka Kinjo
  ANALYTICAL BIOCHEMISTRY, 370, 2, 131, 141, Nov. 2007, [Peer-reviewed]
  English, Scientific journal
• [Prion diseases in animals--bovine spongiform encephalopathy].
  Horiuchi M; Nakamitsu S
  Nihon rinsho. Japanese journal of clinical medicine, 65, 1513, 1520, 8, Aug. 2007, [Peer-reviewed]
• Characterization of prion susceptibility in Neuro2a mouse neuroblastoma cell subclones
  Masahide Uryu; Ayako Karino; Yukiko Kamihara; Motohiro Horiuchi
  MICROBIOLOGY AND IMMUNOLOGY, 51, 7, 661, 669, 2007, [Peer-reviewed], [Corresponding author]
  English, Scientific journal
• Species-specificity of a panel of prion protein antibodies for the immunohistochemical study of animal and human prion disease
  H. Furuoka; A. Yabuzoe; M. Horiuchi; Y. Tagawa; T. Yokoyama; Y. Yamakawa; M. Shinagawa; T. Sata
  JOURNAL OF COMPARATIVE PATHOLOGY, 136, 1, 9, 17, Jan. 2007, [Peer-reviewed]
  English, Scientific journal
• Identification of pH-sensitive regions in the mouse prion by the cysteine-scanning spin-labeling ESR technique
  Yasuko Watanabe; Osamu Inanami; Motohiro Horiuchi; Wakako Hiraoka; Yuhei Shimoyama; Fuyuhiko Inagaki; Mikinori Kuwabara
  BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 350, 3, 549, 556, Nov. 2006, [Peer-reviewed]
  English, Scientific journal
• Inhibition of PrPSc formation by synthetic O-sulfated glycopyranosides and their polymers
  Satoko Yamaguchi; Yoshihiro Nishida; Kenji Sasaki; Mikie Kambara; Chan-Lan Kim; Naotaka Ishiguro; Takehiro Nagatsuka; Hirotaka Uzawa; Motohiro Horiuchi
  BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 349, 2, 485, 491, Oct. 2006, [Peer-reviewed]
  English, Scientific journal
• Distribution of PrPSc in cattle with bovine spongifonn encephalopathy slaughtered at abattoirs in Japan
  N Iwata; Y Sato; Y Higuchi; K Nohtomi; N Nagata; H Hasegawa; M Tobiume; Y Nakamura; K Hagiwara; H Furuoka; M Horiuchi; Y Yamakawa; T Sata
  JAPANESE JOURNAL OF INFECTIOUS DISEASES, 59, 2, 100, 107, Apr. 2006, [Peer-reviewed]
  English, Scientific journal
• Alymphoplasia mice are resistant to prion infection via oral route.
  M Horiuchi; H Furuoka; N Kitamura; M Shinagawa
  JAPANESE JOURNAL OF VETERINARY RESEARCH, 53, 3-4, 149, 157, Feb. 2006, [Peer-reviewed]
  English, Scientific journal
• Sequence variation of bovine prion protein gene in Japanese cattle (Holstein and Japanese black)
  S Nakamitsu; T Miyazawa; M Horiuchi; S Onoe; Y Ohoba; H Kitagawa; N Ishiguro
  JOURNAL OF VETERINARY MEDICAL SCIENCE, 68, 1, 27, 33, Jan. 2006, [Peer-reviewed]
  English, Scientific journal
• [Prion disease as infectious disease transmissible from animals to human].
  Horiuchi M
  Nihon rinsho. Japanese journal of clinical medicine, 63, 12, 2213, 2220, 12, Dec. 2005, [Peer-reviewed]
• Conformational change in full-length mouse prion: A site-directed spin-labeling study
  O Inanami; S Hashida; D Iizuka; M Horiuchi; W Hiraoka; Y Shimoyama; H Nakamura; F Inagaki; M Kuwabara
  BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 335, 3, 785, 792, Sep. 2005, [Peer-reviewed]
  English, Scientific journal
• [Prion diseases as zoonosis].
  Horiuchi M
  Uirusu, 55, 1, 45, 53, 1, Jun. 2005, [Peer-reviewed]
  Japanese, Scientific journal, Prion diseases such as bovine spongiform encephalopathy (BSE) have been recognized as zoonosis since the existence of variant Creutzfeldt-Jakob disease (vCJD) was reported in 1996. BSE became a serious social problem even in Japan after the first BSE case was found in 2001. The incidence of BSE in EU and UK appears declining, and the vCJD incidence also shows a tendency to decrease. On the contrary, fears for the spread of BSE became actual problems: BSE occurrence outside of EU, transmission of vCJD by blood transfusion, and the first vCJD case in Japan. To prevent further spread and to reduce the risk of BSE, it is important to continue BSE screening/surveillance, removal of specified risk materials from food and feed chains, and effective feed regulation. For the disclosure and elimination of prion-contaminated blood, materials for medical and pharmaceutical products and so on, it is required to improve the sensitivity of prion detection methods. Furthermore, it is also important to establish therapeutics of human prion diseases.
• Effective antigen-retrieval method for immunohistochemical detection of abnormal isoform of prion proteins in animals
  H Furuoka; A Yabuzoe; M Horiuchi; Y Tagawa; T Yokoyama; Y Yamakawa; M Shinagawa; T Sata
  ACTA NEUROPATHOLOGICA, 109, 3, 263, 271, Apr. 2005, [Peer-reviewed]
  English, Scientific journal
• Polymorphisms of caprine PrP gene detected in Japan
  Y Kurosaki; N Ishiguro; M Horiuchi; M Shinagawa
  JOURNAL OF VETERINARY MEDICAL SCIENCE, 67, 3, 321, 323, Mar. 2005, [Peer-reviewed]
  English, Scientific journal
• Surveillance of chronic wasting disease in sika deer, Cervus nippon, from Tokachi district in Hokkaido
  N Kataoka; M Nishimura; M Horiuchi; N Ishiguro
  JOURNAL OF VETERINARY MEDICAL SCIENCE, 67, 3, 349, 351, Mar. 2005, [Peer-reviewed]
  English, Scientific journal
• Cell-surface retention of PrPC by anti-PrP antibody prevents protease-resistant PrP formation
  CL Kim; A Karino; N Ishiguro; M Shinagawa; M Sato; M Horiuchi
  JOURNAL OF GENERAL VIROLOGY, 85, 11, 3473, 3482, Nov. 2004, [Peer-reviewed]
  English, Scientific journal
• Unique amino acid polymorphisms of PrP genes in Mongolian sheep breeds
  A Gombojav; N Ishiguro; M Horiuchi; M Shinagawa
  JOURNAL OF VETERINARY MEDICAL SCIENCE, 66, 10, 1293, 1295, Oct. 2004, [Peer-reviewed]
  English, Scientific journal
• Antigenic characterization of an abnormal isoform of prion protein using a new diverse panel of monoclonal antibodies
  CL Kim; A Umetani; T Matsui; N Ishiguro; M Shinagawa; M Horiuchi
  VIROLOGY, 320, 1, 40, 51, Mar. 2004, [Peer-reviewed]
  English, Scientific journal
• Experimental transmission of abnormal prion protein (PrPsc) in the small intestinal epithelial cells of neonatal mice
  M. Okamoto; H. Furuoka; M. Horiuchi; T. Noguchi; K. Hagiwara; Y. Muramatsu; K. Tomonaga; M. Tsuji; C. Ishihara; K. Ikuta; H. Taniyama
  Veterinary Pathology, 40, 6, 723, 727, Nov. 2003, [Peer-reviewed]
  English, Scientific journal
• Cellular prion protein promotes Brucella infection into macrophages
  MH Watarai; S Kim; J Erdenebaatar; S Makino; M Horiuchi; T Shirahata; S Sakaguchi; S Katamine
  JOURNAL OF EXPERIMENTAL MEDICINE, 198, 1, 5, 17, Jul. 2003, [Peer-reviewed]
  English, Scientific journal
• Susceptibility of transgenic mice expressing chimeric sheep, bovine and human PrP genes to sheep scrapie
  A Gombojav; Shimauchi, I; M Horiuchi; N Ishiguro; M Shinagawa; T Kitamoto; Miyoshi, I; S Mohri; M Takata
  JOURNAL OF VETERINARY MEDICAL SCIENCE, 65, 3, 341, 347, Mar. 2003, [Peer-reviewed]
  English, Scientific journal
• Amino acid Polymorphisms of PrP gene in Mongolian sheep
  A Gombojav; N Ishiguro; M Horiuchi; D Serjmyadag; B Byambaa; M Shinagawa
  JOURNAL OF VETERINARY MEDICAL SCIENCE, 65, 1, 75, 81, Jan. 2003, [Peer-reviewed]
  English, Scientific journal
• Bacteriological and genetic assessment of game meat from Japanese wild boars
  Y Naya; M Horiuchi; N Ishiguro; M Shinagawa
  JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, 51, 2, 345, 349, Jan. 2003, [Peer-reviewed]
  English, Scientific journal
• A genetic method to distinguish crossbred inobuta from Japanese wild boars
  N Ishiguro; Y Naya; M Horiuchi; M Shinagawa
  ZOOLOGICAL SCIENCE, 19, 11, 1313, 1319, Nov. 2002, [Peer-reviewed]
  English, Scientific journal
• Biological and biochemical characterization of sheep scrapie in Japan
  M Horiuchi; T Nemoto; N Ishiguro; H Furuoka; S Mohri; M Shinagawa
  JOURNAL OF CLINICAL MICROBIOLOGY, 40, 9, 3421, 3426, Sep. 2002, [Peer-reviewed], [Corresponding author]
  English, Scientific journal
• Multiple nuclear pseudogenes of mitochondrial DNA exist in the canine genome
  N Ishiguro; A Nakajima; M Horiuchi; M Shinagawa
  MAMMALIAN GENOME, 13, 7, 365, 372, Jul. 2002, [Peer-reviewed]
  English, Scientific journal
• Glycidol Degrades Scrapie Mouse Prion Protein
  Mari Yamamoto; Motohiro Horiuchi; Naotaka Ishiguro; Morikazu Shinagawa; Takato Matsuo; Kenji Kaneko
  Journal of Veterinary Medical Science, 63, 9, 983, 990, Japanese Society of Veterinary Science, Sep. 2001, [Peer-reviewed]
  English, Scientific journal
• Inhibition of Interactions and Interconversions of Prion Protein Isoforms by Peptide Fragments from the C-terminal Folded Domain
  Motohiro Horiuchi; Gerald S. Baron; Liang-Wen Xiong; Byron Caughey
  Journal of Biological Chemistry, 276, 18, 15489, 15497, 04 May 2001, [Peer-reviewed]
  English, Scientific journal
• Sulfated glycans and elevated temperature stimulate PrPSc-dependent cell-free formation of protease-resistant prion protein
  C.'n. Wong
  The EMBO Journal, 20, 3, 377, 386, Springer Science and Business Media LLC, 01 Feb. 2001, [Peer-reviewed]
  Scientific journal
• Interactions between heterologous forms of prion protein: Binding, inhibition of conversion, and species barriers
  Motohiro Horiuchi; Suzette A. Priola; Joëlle Chabry; Byron Caughey
  Proceedings of the National Academy of Sciences, 97, 11, 5836, 5841, Proceedings of the National Academy of Sciences, 16 May 2000, [Peer-reviewed], [Lead author]
  Scientific journal, The self-induced formation of the disease-associated, protease-resistant prion protein (PrP-res) from the normal protease-sensitive isoform (PrP-sen) appears to be a key event in the pathogenesis of transmissible spongiform encephalopathies. The amino acid sequence specificity of PrP-res formation correlates with, and may account for, the species specificity in transmission of transmissible spongiform encephalopathy agents in vivo . To analyze the mechanism controlling the sequence specificity of PrP-res formation, we compared the binding of PrP-sen to PrP-res with its subsequent acquisition of protease resistance by using cell-free systems consisting of heterologous versus homologous mouse and hamster PrP isoforms. Our studies showed that heterologous PrP-sen can bind to PrP-res with little conversion to the protease-resistant state and, in doing so, can interfere with the conversion of homologous PrP-sen. The interference occurred with molar ratios of homologous to heterologous PrP-sen molecules as low as 1:1. The interference was due primarily to the inhibition of conversion, but not the binding, of the homologous PrP-sen to PrP-res. The results provide evidence that the sequence specificity of PrP-res formation in this model is determined more by the conversion to protease resistance than by the initial binding step. These findings also imply that after the initial binding, further intermolecular interactions between PrP-sen and PrP-res are required to complete the process of conversion to the protease-resistant state.
• Prion protein interconversions and the transmissible spongiform encephalopathies
  Motohiro Horiuchi; Byron Caughey
  Structure, 7, 10, R231, R240, Elsevier BV, Oct. 1999, [Peer-reviewed], [Lead author]
  English, Scientific journal
• Specific binding of normal prion protein to the scrapie form via a localized domain initiates its conversion to the protease-resistant state
  Motohiro Horiuchi; Byron Caughey
  EMBO Journal, 18, 12, 3193, 3203, 15 Jun. 1999, [Peer-reviewed]
  English, Scientific journal
• Characterization of antibodies raised against bovine-PrP-peptides
  H Takahashi; RH Takahashi; H Hasegawa; M Horiuchi; M Shinagawa; T Yokoyama; K Kimura; M Haritani; T Kurata; K Nagashima
  JOURNAL OF NEUROVIROLOGY, 5, 3, 300, 307, Jun. 1999, [Peer-reviewed]
  English, Scientific journal
• Immunohistochemical study of some cytoskeletal proteins in hereditary myopathy of the diaphragmatic muscles in Holstein-Friesian cattle
  H Furuoka; A Murakami; M Tsuchihashi; H Yokota; T Doi; Y Kobayashi; T Matsui; M Horiuchi; H Taniyama
  ACTA NEUROPATHOLOGICA, 97, 2, 177, 184, Feb. 1999, [Peer-reviewed]
  English, Scientific journal
• [9] Assays of protease-resistant prion protein and its formation
  Byron Caughey; Motohiro Horiuchi; Rémi Demaimay; Gregory J. Raymond
  Methods in Enzymology, 122, 133, Elsevier, 1999, [Peer-reviewed]
  In book
• Detection methods of possible prion contaminants in collagen and gelatin
  T. Nemoto; M. Horiuchi; M. Horiuchi; N. Ishiguro; M. Shinagawa
  Archives of Virology, 144, 177, 184, 01 Jan. 1999, [Peer-reviewed]
• Inhibition of protease-resistant prion protein formation by porphyrins and phthalocyanines
  Winslow S. Caughey; Lynne D. Raymond; Motohiro Horiuchi; Byron Caughey
  Proceedings of the National Academy of Sciences, 95, 21, 12117, 12122, Proceedings of the National Academy of Sciences, 13 Oct. 1998, [Peer-reviewed]
  Scientific journal, A central aspect of pathogenesis in the transmissible spongiform encephalopathies or prion diseases is the conversion of normal protease-sensitive prion protein (PrP-sen) to the abnormal protease-resistant form, PrP-res. Here we identify porphyrins and phthalocyanines as inhibitors of PrP-res accumulation. The most potent of these tetrapyrroles had IC ₅₀ values of 0.5–1 μM in scrapie-infected mouse neuroblastoma (ScNB) cell cultures. Inhibition was observed without effects on protein biosynthesis in general or PrP-sen biosynthesis in particular. Tetrapyrroles also inhibited PrP-res formation in a cell-free reaction composed predominantly of hamster PrP-res and PrP-sen. Inhibitors were found among phthalocyanines, deuteroporphyrins IX, and meso-substituted porphines; examples included compounds containing anionic, neutral protic, and cationic peripheral substituents and various metals. We conclude that certain tetrapyrroles specifically inhibit the conversion of PrP-sen to PrP-res without apparent cytotoxic effects. The inhibition observed in the cell-free conversion reaction suggests that the mechanism involved direct interactions of the tetrapyrrole with PrP-res and/or PrP-sen. These findings introduce a new class of inhibitors of PrP-res formation that represents a potential source of therapeutic agents for transmissible spongiform encephalopathies.
• Differences in the evolutionary pattern of feline panleukopenia virus and canine parvovirus
  Motohiro Horiuchi; Motohiro Horiuchi; Motohiro Horiuchi; Yumi Yamaguchi; Takashi Gojobori; Masami Mochizuki; Hideyuki Nagasawa; Yutaka Toyoda; Naotaka Ishiguro; Morikazu Shinagawa
  Virology, 249, 440, 452, 30 Sep. 1998
• Chicken monoclonal antibodies against synthetic bovine prion protein peptide
  K Matsushita; H Horiuchi; S Furusawa; M Horiuchi; M Shinagawa; H Matsuda
  JOURNAL OF VETERINARY MEDICAL SCIENCE, 60, 6, 777, 779, Jun. 1998
  English, Scientific journal
• Characterization of differentially expressed genes in the bovine T lymphoma cell line
  Hideyuki Onodera; Naotaka Ishiguro; Motohiro Horiuchi; Morikazu Shinagawa
  Veterinary Immunology and Immunopathology, 62, 209, 219, 16 Apr. 1998
• Characterization of the sheep apolipoprotein E (ApoE) gene and allelic variations of the ApoE gene in scrapie Suffolk sheep
  Yusuke Komatsu; Motohiro Horiuchi; Motohiro Horiuchi; Naotaka Ishiguro; Takane Matsui; Morikazu Shinagawa
  Gene, 208, 2, 131, 138, Elsevier BV, 22 Feb. 1998, [Peer-reviewed]
  Scientific journal
• Genomic structure of the bovine PrP gene and complete nucleotide sequence of bovine PrP cDNA
  M Horiuchi; N Ishiguro; H Nagasawa; Y Toyoda; M Shinagawa
  ANIMAL GENETICS, 29, 1, 37, 40, Feb. 1998, [Peer-reviewed], [Lead author]
  English, Scientific journal
• Virus Inactivation in Superoxide Dismutase Preparations by Ultraviolet Light Irradiation.
  Junichi HIRAYAMA; Hideki ABE; Kenji IKEBUCHI; Motohiro HORIUCHI; Morikazu SHINAGAWA; Naoki KAMO; Sadayoshi SEKIGUCHI
  Biological and Pharmaceutical Bulletin, 21, 6, 621, 623, Pharmaceutical Society of Japan, 1998, [Peer-reviewed]
  Scientific journal
• Photoinactivation of Virus Infectivity by Hypocrellin A
  Junichi Hirayama; Kenji Ikebuchi; Hideki Abe; Kil‐Won Kwon; Yoshiko Ohnishi; Motohiro Horiuchi; Morikazu Shinagawa; Kazuyoshi Ikuta; Naoki Kamo; Sadayoshi Sekiguchi
  Photochemistry and Photobiology, 66, 5, 697, 700, Wiley, Nov. 1997, [Peer-reviewed]
  Scientific journal, Abstract— We investigated the photoinactivation of virus infectivity by hypocrellin A and its mechanism. The titers of vesicular stomatitis virus (VSV) and human immunodeficiency virus type 1 (HIV‐1), both of which are enveloped viruses, were reduced upon illumination with hypocrellin A in a concentration‐dependent manner, whereas canine parvovirus, a nonenveloped virus, was not killed. The removal of oxygen or addition of sodium azide or bT‐carotene both inhibited VSV inactivation. Mannitol and superoxide dismutase had no effect on VSV inactivation. These results indicate that singlet oxygen was involved in the process of VSV inactivation. Of the three major VSV membrane proteins, peripheral membrane protein M was most damaged by the hypocrellin A phototreatment.
• Instability of the 12-nucleotide repeat in c-myb gene of bovine T-lymphoma cells
  T Shinagawa; N Ishiguro; M Horiuchi; M Shinagawa
  JOURNAL OF VETERINARY MEDICAL SCIENCE, 59, 11, 1071, 1074, Nov. 1997
  English, Scientific journal
• Deletion of c-myb exon 9 induced by insertion of repeats
  Toshie Shinagawa; Naotaka Ishiguro; Motohiro Horiuchi; Takane Matsui; Kosuke Okada; Morikazu Shinagawa
  Oncogene, 14, 23, 2775, 2783, 07 Aug. 1997, [Peer-reviewed]
• Alternative usage of exon 1 of bovine PrP mRNA
  Motohiro Horiuchi; Motohiro Horiuchi; Naotaka Ishiguro; Hideyuki Nagasawa; Yutaka Toyoda; Morikazu Shinagawa
  Biochemical and Biophysical Research Communications, 233, 650, 654, 28 Apr. 1997, [Lead author]
• Sensitive enzyme-linked immunosorbent assay for detection of PrP(Sc) in crude tissue extracts from scrapie-affected mice
  Kai Uwe; D Grathwohl; Motohiro Horiuchi; Motohiro Horiuchi; Naotaka Ishiguro; Morikazu Shinagawa
  Journal of Virological Methods, 64, 205, 216, 01 Mar. 1997, [Peer-reviewed]
• p53 mutation as a potential cellular factor for tumor development in enzootic bovine leukosis
  Naotaka Ishiguro; Hidefumi Furuoka; Takane Matsui; Motohiro Horiuchi; Morikazu Shinagawa; Masatoshi Asahina; Kousuke Okada
  Veterinary Immunology and Immunopathology, 55, 351, 358, 01 Mar. 1997, [Peer-reviewed]
• Characterization of the bovine prion protein gene: The expression requires interaction between the promoter and intron
  S Inoue; M Tanaka; M Horiuchi; N Ishiguro; M Shinagawa
  JOURNAL OF VETERINARY MEDICAL SCIENCE, 59, 3, 175, 183, Mar. 1997, [Peer-reviewed]
  English, Scientific journal
• Epitope mapping of a monoclonal antibody specific to feline panleukopenia virus and mink enteritis virus
  M Horiuchi; M Mochizuki; N Ishiguro; H Nagasawa; M Shinagawa
  JOURNAL OF VETERINARY MEDICAL SCIENCE, 59, 2, 133, 136, Feb. 1997, [Peer-reviewed], [Lead author]
  English, Scientific journal
• Characterization of differentiation antigens expressed in bovine lymphosarcomas
  Y Sasaki; N Ishiguro; M Horiuchi; M Shinagawa; S Osame; H Furuoka; T Matsui; M Asahina; K Okada
  JOURNAL OF COMPARATIVE PATHOLOGY, 116, 1, 13, 20, Jan. 1997, [Peer-reviewed]
  English, Scientific journal
• Differentiation of vaccine virus from field isolates of feline panleukopenia virus by polymerase chain reaction and restriction fragment length polymorphism analysis
  Motohiro Horiuchi; Kazuyo Yuri; Takehisa Soma; Hiromi Katae; Hideyuki Nagasawa; Morikazu Shinagawa
  Veterinary Microbiology, 53, 3-4, 283, 293, Dec. 1996, [Peer-reviewed], [Lead author]
  English, Scientific journal
• Isolation of canine parvovirus from a cat manifesting clinical signs of feline panleukopenia
  M Mochizuki; M Horiuchi; H Hiragi; M C San Gabriel; N Yasuda; T Uno
  Journal of Clinical Microbiology, 34, 9, 2101, 2105, American Society for Microbiology, Sep. 1996, [Peer-reviewed]
  Scientific journal, Twenty-seven feline parvovirus (FPV) isolates were recovered from cats clinically diagnosed with feline panleukopenia (FPL) for assessing antigenic and genomic properties of FPL viruses (FPLV) recently prevalent among cats in Japan. All isolates, with the exception of one novel isolate, FPV-314, possessed homologous properties, and their subgroups in FPVs were identified as FPLV. The FPV-314 isolate, which was from a 1.5-year-old cat which manifested clinical signs of FPL and died on the 13th day after the first medical examination, was finally identified as canine parvovirus (CPV) because it lacked a specific antigenic epitope commonly detected in FPLV and mink enteritis virus and because the nucleotide sequence of the capsid protein gene was almost identical to those of CPV-2a and -2b antigenic type strains recently prevalent among dogs in Japan. The present result together with our previous findings (M. Mochizuki, R. Harasawa, and H. Nakatani. Vet. Microbiol. 38:1-10, 1993) indicates the possibility that CPV and FPLV undergo mutual interspecies transmission between dogs and cats, and it is postulated that they may cause disease in some adventitious hosts.
• Predominant p53 mutations in enzootic bovine leukemic cell lines
  Hiroshi Komori; Naotaka Ishiguro; Motohiro Horiuchi; Morikazu Shinagawa; Yoko Aida
  Veterinary Immunology and Immunopathology, 52, 53, 63, 15 Jun. 1996, [Peer-reviewed]
• Potential transforming-ability by internal deletion of the bovine c-myb gene
  T Ohozono; N Ishiguro; M Horiuchi; M Shinagawa
  JOURNAL OF VETERINARY MEDICAL SCIENCE, 58, 4, 305, 310, Apr. 1996, [Peer-reviewed]
  English, Scientific journal
• Improvement of PrPSc-detection in mouse spleen early at the preclinical stage of scrapie with collagenase-completed tissue homogenization and Sarkosyl-NaCl extraction of PrPSc
  KUD Grathwohl; M Horiuchi; N Ishiguro; M Shinagawa
  ARCHIVES OF VIROLOGY, 141, 10, 1863, 1874, 1996, [Peer-reviewed]
  English, Scientific journal
• BOVINE HERPESVIRUS MENINGOENCEPHALITIS ASSOCIATION WITH INFECTIONS BOVINE-RHINOTRACHEITIS (IBR) VACCINE
  H FURUOKA; N IZUMIDA; M HORIUCHI; S OSAME; T MATSUI
  ACTA NEUROPATHOLOGICA, 90, 6, 565, 571, Dec. 1995, [Peer-reviewed]
  English, Scientific journal
• A CELLULAR-FORM OF PRION PROTEIN (PRPC) EXISTS IN MANY NONNEURONAL TISSUES OF SHEEP
  M HORIUCHI; N YAMAZAKI; T IKEDA; N ISHIGURO; M SHINAGAWA
  JOURNAL OF GENERAL VIROLOGY, 76, 2583, 2587, Oct. 1995, [Peer-reviewed], [Lead author]
  English, Scientific journal
• AMINO-ACID POLYMORPHISMS OF PRP WITH REFERENCE TO ONSET OF SCRAPIE IN SUFFOLK AND CORRIEDALE SHEEP IN JAPAN
  T IKEDA; M HORIUCHI; N ISHIGURO; Y MURAMATSU; GD KAIUWE; M SHINAGAWA
  JOURNAL OF GENERAL VIROLOGY, 76, 2577, 2581, Oct. 1995, [Peer-reviewed]
  English, Scientific journal
• RESTRICTION-ENDONUCLEASE ANALYSIS OF BOVINE HERPESVIRUS TYPE-1 ISOLATES FROM CALVES WITH FATAL ENCEPHALITIS - COMPARISON WITH VACCINE VIRUS
  M HORIUCHI; N YAMAZAKI; H FURUOKA; T MATSUI; M NAKAGAWA; N ISHIGURO; M SHINAGAWA
  JOURNAL OF VETERINARY MEDICAL SCIENCE, 57, 3, 577, 580, Jun. 1995, [Peer-reviewed], [Lead author]
  English, Scientific journal
• A SPONTANEOUS INTERNAL DELETION OF THE C-MYB PROTOONCOGENE ENHANCES TRANSCRIPTIONAL ACTIVATION IN BOVINE T-LYMPHOMA-CELLS
  N ISHIGURO; T OHZONO; T SHINAGAWA; M HORIUCHI; M SHINAGAWA
  JOURNAL OF BIOLOGICAL CHEMISTRY, 269, 43, 26822, 26829, Oct. 1994, [Peer-reviewed]
  English, Scientific journal
• CHARACTERIZATION OF MONOCLONAL-ANTIBODIES AGAINST SPORADIC BOVINE LEUKOSIS CELL-LINES
  T SHINAGAWA; N ISHIGURO; M HORIUCHI; M SHINAGAWA; T MATSUI
  JOURNAL OF VETERINARY MEDICAL SCIENCE, 56, 5, 827, 833, Oct. 1994, [Peer-reviewed]
  English, Scientific journal
• MONOCLONAL-ANTIBODY S37 INDUCES APOPTOSIS IN BOVINE T-LYMPHOMA CELLS
  T SHINAGAWA; N ISHIGURO; M HORIUCHI; M SHINAGAWA
  JOURNAL OF VETERINARY MEDICAL SCIENCE, 56, 5, 957, 959, Oct. 1994, [Peer-reviewed]
  English
• SURVEY OF NATURAL SCRAPIE IN JAPAN - ANALYSIS OF RFLP TYPES OF THE PRP GENE AND DETECTION OF PRPSC MAINLY IN SUFFOLK SHEEP
  A ONODERA; T IKEDA; M HORIUCHI; N ISHIGURO; M ONUMA; N HIRANO; T MIKAMI; E HONDA; K HIRAI; K KAI; H YUGI; Y MURAMATSU; M SHINAGAWA
  JOURNAL OF VETERINARY MEDICAL SCIENCE, 56, 4, 627, 632, Aug. 1994, [Peer-reviewed]
  English, Scientific journal
• MAPPING OF DETERMINANTS OF THE HOST-RANGE FOR CANINE CELLS IN THE GENOME OF CANINE PARVOVIRUS USING CANINE PARVOVIRUS MINK ENTERITIS VIRUS CHIMERIC VIRUSES
  M HORIUCHI; H GOTO; N ISHIGURO; M SHINAGAWA
  JOURNAL OF GENERAL VIROLOGY, 75, 1319, 1328, Jun. 1994, [Peer-reviewed], [Lead author]
  English, Scientific journal
• THE SIGNIFICANCE OF PRPSC DETECTION FOR THE DIAGNOSIS OF INSIDIOUS SCRAPIE
  Y MURAMATSU; A ONODERA; M HORIUCHI; N ISHIGURO; M SHINAGAWA
  SLOW INFECTIONS OF THE CENTRAL NERVOUS SYSTEM, 724, 347, 349, 1994, [Peer-reviewed]
  English
• DETECTION OF PRP(SC) IN SHEEP AT THE PRECLINICAL STAGE OF SCRAPIE AND ITS SIGNIFICANCE FOR DIAGNOSIS OF INSIDIOUS INFECTION
  Y MURAMATSU; A ONODERA; M HORIUCHI; N ISHIGURO; M SHINAGAWA
  ARCHIVES OF VIROLOGY, 134, 3-4, 427, 432, 1994, [Peer-reviewed]
  English
• FAMILIAL SCRAPIE CASES WITH SHORTENED INCUBATION PERIODS
  M SHINAGAWA; Y MURAMATSU; A ONODERA; T MATSUI; M HORIUCHI; N ISHIGURO; M NAKAGAWA
  JOURNAL OF VETERINARY MEDICAL SCIENCE, 55, 4, 665, 667, Aug. 1993, [Peer-reviewed]
  English, Scientific journal
• CONSTRUCTION OF AN INFECTIOUS DNA CLONE OF THE Y-1 STRAIN OF CANINE PARVOVIRUS AND CHARACTERIZATION OF THE VIRUS DERIVED FROM THE CLONE
  M HORIUCHI; M SHINAGAWA
  ARCHIVES OF VIROLOGY, 130, 3-4, 227, 236, 1993, [Peer-reviewed]
  English, Scientific journal
• Comparative sequence analysis and expression of bovine PrP gene in mouse L-929 cells
  Jun Yoshimoto; Toshiyuki Iinuma; Naotaka Ishiguro; Motohiro Horiuchi; Masakatu Imamura; Morikazu Shinagawa
  Virus Genes, 6, 4, 343, 356, Kluwer Academic Publishers, Nov. 1992, [Peer-reviewed]
  English, Scientific journal
• CHARACTERIZATION OF THE STAGE(S) IN THE VIRUS-REPLICATION CYCLE AT WHICH THE HOST-CELL SPECIFICITY OF THE FELINE PARVOVIRUS SUBGROUP IS REGULATED IN CANINE CELLS
  M HORIUCHI; N ISHIGURO; H GOTO; M SHINAGAWA
  VIROLOGY, 189, 2, 600, 608, Aug. 1992, [Peer-reviewed]
  English, Scientific journal
• Biological activity of subfractions from scrapie-associated fibrils
  Sasaki Kazuhiko; Tanaka Koji; Ito Masako; Isomura Hiroshi; Horiuchi Motohiro; Ishiguro Naotaka; Shinagawa Morikazu
  Virus Research, 23, 3, 241, 251, Elsevier BV, May 1992, [Peer-reviewed]
  Scientific journal
• A SPECIFIC RFLP TYPE ASSOCIATED WITH THE OCCURRENCE OF SHEEP SCRAPIE IN JAPAN
  Y MURAMATSU; K TANAKA; M HORIUCHI; N ISHIGURO; M SHINAGAWA; T MATSUI; T ONODERA
  ARCHIVES OF VIROLOGY, 127, 1-4, 1, 9, 1992, [Peer-reviewed]
  English, Scientific journal
• Construction and nucleotide sequence analysis of an infectious DNA clone of the autonomous parvovirus, mink enteritis virus.
  T Kariatsumari; M Horiuchi; E Hama; K Yaguchi; N Ishigurio; H Goto; M Shinagawa
  The Journal of general virology, 72 ( Pt 4), 867, 75, Apr. 1991, [International Magazine]
  English, Scientific journal, We have cloned the replicative form (RF-) DNA of mink enteritis virus (MEV), constructed an infectious recombinant plasmid containing MEV DNA and determined the nucleotide sequence of the cloned MEV DNA. RF-DNAs were detected and infectious virus was generated when the recombinant plasmid containing the entire MEV genome was introduced into feline kidney cell cultures. The MEV genome was 5094 nucleotides (nt) in length; the 3' end of the virion strand contained a 205 nt palindromic sequence and the 5' end a 62 nt palindromic sequence that could assume Y- and U-shaped configurations, respectively. The 5' end of the virion strand had a direct repeat of 61 nt at the carboxyl terminus of the capsid protein gene. The organization of the MEV genome is similar to those of canine parvovirus (CPV) and feline panleukopenia virus (FPLV); there are two large open reading frames (ORFs), one in the 3' half and the other in the 5' half of the genome, with coding capacities of 668 and 722 amino acid residues, respectively. Both are in the same reading frame and no significant ORFs are apparent in the virion strand (negative-sense strand). Possible functional promoter motifs are located at map unit (m.u.) 4.5 and m.u. 40, and a possible functional poly(A) signal is located at m.u. 96. The nucleotide and amino acid sequence homology with CPV and FPLV is greater than 98%, consistent with the hypothesis that MEV and CPV are host-range variants of FPLV.
• IDENTIFICATION OF VIRUS-SPECIFIC ANTIGENS IN CULTURED-CELLS INFECTED WITH MAREKS-DISEASE VIRUS SEROTYPE-2 AND CVI-988 STRAIN
  M HORIUCHI; H KODAMA; T MIKAMI
  JAPANESE JOURNAL OF VETERINARY SCIENCE, 52, 5, 985, 994, Oct. 1990, [Peer-reviewed]
  English, Scientific journal
• CHARACTERIZATION OF REPLICATIVE FORM DNA OF THE AUTONOMOUS PARVOVIRUS MINK ENTERITIS VIRUS
  M SHINAGAWA; Y NOMURA; T KARIATUMARI; N ISHIGURO; M HORIUCHI; H GOTO
  MICROBIOLOGY AND IMMUNOLOGY, 33, 9, 721, 732, 1989, [Peer-reviewed]
  English, Scientific journal
• Isolation of ortho- and paramyxoviruses from migrating feral ducks in Japan
  T. Mikami; M. Kawamura; T. Kondo; T. Murai; M. Horiuchi; H. Kodama; H. Izawa; H. Kida
  Veterinary Record, 120, 17, 417, 418, Wiley, 25 Apr. 1987, [Peer-reviewed]
  Scientific journal

• 標準的感染症対策プログラムは動物病院において手指衛生コンプライアンス向上と多剤耐性菌出現抑制に貢献する
  笹岡 一慶; 森下 啓太郎; 細谷 謙次; 佐藤 敬近; 佐藤 豊孝; 堀内 基広; 滝口 満喜, 日本獣医学会学術集会講演要旨集, 166回, 174, 174, Sep. 2023
  (公社)日本獣医学会, Japanese
• プリオン病治療実験モデル系確立の試み-免疫療法と細胞治療の可能性-
  堀内基広, 臨床評価, 44, 4, 2017
• MALDI-TOF質量分析システムによる犬口腔内からのBergeyella zoohelcumの検出
  原谷那美; 内田玲麻; 郡山尚紀; 鈴木章夫; 堀内基広; 村松康和, 北海道獣医師会雑誌, 59, 8, 2015
• プリオンの感染と細胞内動態
  山崎剛士; 堀内基広, Dement Jpn, 27, 2, 215, 224, 2013
  Japanese
• 異常型プリオンタンパク質の細胞内増殖
  堀内基広; 山崎剛士, Dementia Japan, 25, 3, 2011
• 人獣共通感染症の克服に向けて プリオン病
  堀内基広, 最新医学, 66, 12, 2713, 2720, 2011
  大阪 : 最新医学社, Japanese
• プリオンの増殖とその抑制
  堀内 基広, ウイルス感染症セミナー 10, 13, 25, 2008
• Bovine spongiform encephalopathy
  NAKAMITSU Satoshi; HORIUCHI Motohiro, 臨床とウイルス, 35, 4, 301, 310, 10 Oct. 2007
  Japanese
• Prion diseases in animals: bovine spongiform encephalopathy
  堀内基広; 中満智史, Japanese journal of clinical medicine, 65, 8, 1513, 1520, 2007
  日本臨床社, Japanese
• Prion
  HORIUCHI Motohiro, 臨床とウイルス, 35, 4, 293, 300, 2007
  Japanese
• 北海道のウシにおけるプリオン遺伝子の多形性
  村松康和; 堀内基広; 田村豊, 日本獣医学会学術集会講演要旨集, 141st, 2006
• 血漿分画製剤のろ過工程における異常プリオン除去に関する検討
  前野英毅; 川畑嘉之; 室塚剛志; 脇坂明美; 谷藤正明; 堀内基広; 伴野丞計, 血液事業, 29, 2, 2006
• 小沢義博氏「アメリカのBSE対策の現状」に述べられた21ヵ月齢牛の判定に関するコメントに対する反論
  北本 哲之; 佐多 徹太郎; 品川 森一; 古岡 秀文; 堀内 基弘; 山河 芳夫; 山内 一也; 横山 隆, 67, 11, J9, J10, 25 Nov. 2005
  社団法人日本獣医学会, Japanese
• BSE診断法の開発と現状 (特集 プリオン病)
  堀内 基広, Virus report, 2, 1, 20, 27, Jun. 2005
  医薬ジャーナル社, Japanese
• ニワトリモノクローナル抗体を用いた異常プリオン蛋白質の検出
  青笹正義; 宮本和慶; 西道教尚; 木村聡太; 品川森一; 佐多徹太郎; 堀内基広; 横山隆; 堀内浩幸; 古沢修一; 松田治男, 臨床病理, 53, 2005
• 動物由来感染症としてのプリオン病
  堀内基広, 日本臨床, 63, 12, 2213, 2220, 2005
• Biosynthesis and Transmission of Abnormal Prion Protein
  堀内基広, 膜, 30, 2, 78, 83, 2005
  THE MEMBRANE SOCIETY OF JAPAN, Japanese
• Prion diseases as zoonosis
  堀内基広, ウイルス, 55, 1, 45, 53, 2005
  日本ウィルス学会, Japanese
• 教室紹介 北海道大学大学院獣医学研究科プリオン病学講座
  堀内 基広, ウイルス = Virus / 日本ウィルス学会 編, 54, 2, 267, 269, Dec. 2004
  東京 : 日本ウィルス学会, Japanese
• プリオンの部位特異的スピンラベル法(SDSL-ESR)による構造変化の解析
  稲波修; 橋田修吉; 飯塚大輔; 井上哲; 堀内基広; 桑原幹典, 日本獣医学会学術集会講演要旨集, 138th, 2004
• Inhibitory Activity of Sulfonated GlcNAc Derivatives against Sialidases and Scrapie Prions
  佐々木健二; 西田芳弘; 神原実季恵; 堀内基広; 鵜沢浩隆; 小林一清, 日本化学会講演予稿集, 84th, 2, 2004
• 現行の牛海綿状脳症検査のシカ慢性消耗病(CWD)診断への応用性
  嶋田希実; 岩丸祥史; 林浩子; 高田益宏; 牛木祐子; 木村久美子; 田川裕一; 堀内基広; 品川森一, 日本獣医学会学術集会講演要旨集, 137th, 2004
• 人獣共通感染症としてのプリオン病
  堀内基広, 日本ウイルス学会学術集会プログラム・抄録集, 52nd, 2004
• 抗PrP抗体によるPrP^Sc産生抑制機構の解析
  キム チャンラン; 堀内基広; 石黒直隆; 品川森一, 日本獣医学会学術集会講演要旨集, 138th, 2004
• Site-directed spin label(SDSL)法によるマウスプリオンタンパク質の銅イオン結合部位周辺の構造解析
  稲波修; 橋田修吉; 飯塚大輔; 堀内基広; 平岡和佳子; 稲垣冬彦; 桑原幹典, 電子スピンサイエンス学会年会講演要旨集, 43rd, 2004
• プリオンの環境汚染評価-植物への影響
  三隅智子; 石黒直隆; 堀内基広; 品川森一, 日本獣医学会学術集会講演要旨集, 135th, 2003
• プリオン蛋白質と結合するペプチド性リガンドの探索
  大林浩二; 堀内基広; 石黒直隆; 品川森一, 日本獣医学会学術集会講演要旨集, 135th, 2003
• 抗PrPモノクローナル抗体パネルによるPrPの構造解析
  金チャンラン; 堀内基広; 石黒直隆; 品川森一, 日本獣医学会学術集会講演要旨集, 135th, 2003
• 尿崩症を誘発するマウス馴化スクレイピー株の分離と解析
  田村勇耕; 堀内基広; 石黒直隆; 古岡秀文; 品川森一, 日本獣医学会学術集会講演要旨集, 135th, 2003
• Improvement in sensitivity of PrP^Sc detection method using anti-PrP antibody
  堀内基広; 工藤聡子; 品川森一; 梅谷淳, プリオン病及び遅発性ウイルス感染に関する調査研究 平成14年度研究報告書, 2003
• BSEスクリーニング用ELISA(OFR ELISA)の開発とその性能評価
  堀内基広; 梅谷淳; 工藤聡子; 石黒直隆; 横山隆; 品川森一, 日本獣医学会学術集会講演要旨集, 135th, 2003
• 動物プリオン病の免疫組織化学的染色における抗原賦活化法の検討
  薮添敦史; 古岡秀文; 堀内基広; 田川裕一; 横山隆; 品川森一; 佐多徹太郎, 日本獣医学会学術集会講演要旨集, 135th, 2003
• 抗体療法-基礎から臨床へ- 抗PrP抗体によるプリオン増殖抑制とプリオン病治療の可能性
  堀内基広, 最新医学, 58, 12, 2802, 2808, 2003
• プリオン病-最近の知見 牛海綿状脳症問題に関する最近の動向
  堀内基広, 老年精神医学雑誌, 14, 12, 1488, 1494, 2003
  ワールドプランニング, Japanese
• 経口ルートによるプリオンの感染成立には消化管リンパ装置の存在が必要である
  堀内基広; 石黒直隆; 品川森一; 古岡秀文; 北村延夫, 日本獣医学会学術集会講演要旨集, 133rd, 2002
• プリオン病とその診断 プリオンの検出技術
  堀内基広, 臨床検査, 46, 12, 1545, 1551, 2002
  株式会社医学書院
• 感染症 プリオン病 病態の解明を目指して プリオン蛋白質の構造と遺伝子多型
  堀内基広, Mebio, 19, 12, 2002
• プリオン蛋白質とプリオン病
  堀内基広, 栄養生理研究会報, 46, 1, 45, 50, 2002
  札幌 : 家畜栄養生理研究会, Japanese
• Inspection of Scrapie Prion Contamination in Stock Powder of Gastropylore.
  品川森一; 堀内基広; 石黒直隆, 医薬品研究, 33, 7, 2002
• 牛海綿状脳症に関する検査概要と今後の対応
  池田 徹也; 堀内 基広; 古岡 秀文, 食品衛生研究, 52, 1, 33, 42, Jan. 2002
  日本食品衛生協会, Japanese
• モンゴルの羊におけるプリオン蛋白質のアミノ酸多型
  ゴンボジャブ アルタンゲレル; 堀内基広; 石黒直隆; 品川森一, 日本獣医学会学術集会講演要旨集, 131st, 2001
• Prion detection by immunological methods
  SHINAGAWA Morikazu; HORIUCHI Motohiro; MATSUI Takane, SEIKATSU EISEI (Journal of Urban Living and Health Association), 45, 5, 259, 269, 2001
  Osaka Urban Living and Health Association
• Prion diseases in animals
  HORIUCHI Motohiro, Virus, 51, 2, 145, 150, 2001
  動物のプリオン病は全て感染因子プリオンが体外から侵入することによりおこる感染性プリオン病である. そのうち, 羊のスクレイピーおよび鹿の慢性消耗病は自然状態でそれぞれの宿主間で感染が成立する. この2種以外の動物プリオン病の発生には人為的要素が関与している. 中枢神経系における異常型プリオン蛋白質PrP^Scの蓄積, 神経細胞の空胞変性など, 特徴的な所見は共通であるが, プリオンの体内分布など相違点も明らかになってきた. プリオンの侵入門戸および中枢神経系組織への侵入経路なども解明されつつある. 本稿では, 動物プリオン病の発生原因, プリオンの感染経路および伝播様式, 株の多様性等について, 主に羊スクレイピー, BSEを研究対象として得られた知見を紹介する. また, 日本のBSE検査についても紹介する., The Japanese Society for Virology, English
• Sheep biodistribution of scrapie prion protein. PrPSc detection by Western blotting.
  高橋未緒; 堀内基広; 石黒直隆; 松井高峯; 古岡秀文; 品川森一, 日本獣医学会学術集会講演要旨集, 126th, 1998
• Distribution of scrapie prion protein in sheep body. Immunohistochemical retrieval by anti-PrP antibody.
  鈴木智; 高橋広志; 松井高峯; 堀内基広; 石黒直隆; 品川森一; 古林与志安; 古岡秀文, 日本獣医学会学術集会講演要旨集, 126th, 1998
• Prion disease.
  堀内基広, 食肉の科学, 38, 2, 205, 212, 1997
  資料形態 : テキストデータ プレーンテキスト
  コレクション : 国立国会図書館デジタルコレクション > デジタル化資料 > 雑誌, 日本食肉研究会, Japanese
• Analysis of cattle prion protein PrP gene promoter region.
  井上真吾; 田中成幸; 堀内基広; 石黒直隆; 品川森一, 日本獣医学会学術集会講演要旨集, 122nd, 1996
• Metabolism and distribution of prion protein.
  品川森一; 堀内基広, 実験医学, 14, 18, 1996
• プリオン病の分子遺伝学と生物物理学
  Prusiner Stanley B; 堀内 基広; 品川 森一, 蛋白質核酸酵素, 40, 16, p2383, 2407, Dec. 1995
  記事分類: 生物学--生理・生化学・生物物理--核酸・蛋白質・酵素 ; 医学--精神神経科学, 共立出版, Japanese
• ANALYSIS OF MAREK'S DISEASE VIRUS SEROTYPE 1-SPECIFIC, SEROTYPE 2-SPECIFIC AND CROSS-REACTIVE POLYPEPTIDES IN VIRUS-INFECTED CELLS
  HORIUCHI Motohiro, Japanese journal of veterinary research, 36, 2, 150, 150, 20 May 1988
  Hokkaido University, English

• プリオン感染マウスの中枢神経組織で見られる領域特異的な神経細胞死
  星加恭; 小松勇介; 小松勇介; TEMUULEN Erdenebat; 鈴木章夫; 佐藤豊孝; 佐藤豊孝; 堀内基広; 堀内基広
  日本Cell Death学会学術集会プログラム抄録集, 2023
  2023 - 2023
• Two distinct cell death process are induced in the central nerve tissue of prion-infected mice
  星加恭; 星加恭; 小松勇介; 小松勇介; ERDENEBAT Temuulen; ERDENEBAT Temuulen; 鈴木章夫; 佐藤豊孝; 佐藤豊孝; 堀内基広; 堀内基広
  日本ウイルス学会学術集会プログラム・予稿集(Web), 2023
  2023 - 2023
• Two distinct neuronal cell death processes, apoptosis and ferroptosis, are observed in prion infected mice
  星加恭; 星加恭; 小松勇介; ERDENEBAT Temuulen; ERDENEBAT Temuulen; 鈴木章夫; 佐藤豊隆; 堀内基広
  日本分子生物学会年会プログラム・要旨集(Web), 2022
  2022 - 2022
• -各分野で活躍する獣医師のさらなる飛躍に向けて(V)-国際的な獣医学教育第三者評価機関の認証取得とわが国の獣医学教育改革
  堀内基広
  日本獣医師会雑誌, 2022
  2022 - 2022
• プリオン22L株のグルタミン酸作動性神経細胞における易増殖性
  上森望未; 田中美咲; TEMUULEN Erdenebat; 小松勇介; 鈴木章夫; 堀内基広
  日本獣医学会学術集会講演要旨集, 2021
  2021 - 2021
• プリオン感染動物で見られるシナプス脱落についての比較病理学的検討
  星加恭; 星加恭; 能瀬絵菜; 小松勇介; 堀内基広; 古岡秀文
  日本獣医学会学術集会講演要旨集, 2021
  2021 - 2021
• プリオン感染初代培養神経細胞における神経細胞自律的変化の解析
  田中美咲; 山崎剛士; 長谷部理絵; 鈴木章夫; 堀内基広
  日本獣医学会学術集会講演要旨集, 2020
  2020 - 2020
• Real-time quaking-induced conversion(RT-QuIC)によるCWDおよび非定型BSE検出における組換えシカプリオンタンパク質の有用性
  鈴木章夫; 澤田和平; 山崎剛士; 堀内基広
  日本獣医学会学術集会講演要旨集, 2020
  2020 - 2020
• 異常型プリオンタンパク質生成がニューロンに及ぼす直接的神経変性効果について
  田中美咲; 山崎剛士; 鈴木章夫; 長谷部理絵; 堀内基広
  日本分子生物学会年会プログラム・要旨集(Web), 2019
  2019 - 2019
• 神経細胞特異的遺伝子発現解析によるプリオン病の神経変性機構の解析
  島倉綾乃; 山崎剛士; 鈴木章夫; 長谷部理絵; 堀内基広
  日本獣医学会学術集会講演要旨集, 2018
  2018 - 2018
• 定型BSEおよび非定型BSEプリオンに感染したマウスの脳のトランスクリプトーム解析
  山崎剛士; 岩丸祥史; 鈴木章夫; 長谷部理絵; 堀内基広
  日本獣医学会学術集会講演要旨集, 2018
  2018 - 2018
• 次世代シーケンサーを用いた,プリオン感染マウス脳由来ミクログリアおよびアストロサイトの活性化状態の解析
  黒田弥乃梨; 山崎剛士; 鈴木章夫; 長谷部理絵; 堀内基広
  日本獣医学会学術集会講演要旨集, 2016
  2016 - 2016
• 異常型プリオンタンパク質(PrP^Sc)特異的染色法によるプリオン持続感染株化細胞とプリオン感染初代培養神経細胞におけるPrP^Scの解析
  田中美咲; 舛甚賢太郎; 舛甚賢太郎; 山崎剛士; 長谷部理絵; 鈴木章夫; 堀内基広
  日本獣医学会学術集会講演要旨集, 2016
  2016 - 2016
• イヌ口腔内からのBergeyella zoohelcumの分離
  村松康和; 原谷那美; 内田玲麻; 郡山尚紀; 鈴木章夫; 堀内基広
  日本獣医学会学術集会講演要旨集, 2015
  2015 - 2015
• BSEおよびL型BSEの異種間交差伝達実験によるプリオン株の病理学的特徴への種差の影響に関する検討
  榎芙蓉; 木元美樹; 室井喜景; 堀内基広; 古岡秀文
  日本獣医学会学術集会講演要旨集, 2015
  2015 - 2015
• 次世代シーケンサーによるリステリア菌国内分離株のゲノム比較解析
  長谷部理絵; 中尾亮; 平井佑治; 鈴木章夫; 山崎剛士; 澤洋文; 高井伸二; 堀内基広
  日本獣医学会学術集会講演要旨集, 2014
  2014 - 2014
• 補体反応はプリオン感染マウス大脳皮質由来初代培養神経細胞の細胞膜透過性を亢進し,異常型プリオンタンパク質の蓄積量を変化させる
  長谷部理絵; 鈴木章夫; 山崎剛士; 堀内基広
  日本ウイルス学会学術集会プログラム・抄録集, 2014
  2014 - 2014
• プリオン病の病態進行に伴うアストロサイト活性化状態の解析
  黒田弥乃梨; 山崎剛士; 鈴木章夫; 長谷部理絵; 堀内基広
  日本ウイルス学会学術集会プログラム・抄録集, 2014
  2014 - 2014
• プリオン感染マウス脳由来アストロサイトの活性化状態の解析
  黒田弥乃梨; 山崎剛士; 鈴木章夫; 長谷部理絵; 堀内基広
  日本獣医学会学術集会講演要旨集, 2014
  2014 - 2014
• 大脳皮質由来初代培養神経細胞におけるプリオン感染の解析
  藤原愛; 山崎剛士; 鈴木章夫; 長谷部理絵; 堀内基広
  日本獣医学会学術集会講演要旨集, 2014
  2014 - 2014
• 北米エルクに由来するCWDプリオンの伝達試験に関する研究
  綿貫亨; 室井喜景; 横山隆; 堀内基広; 古岡秀文
  日本獣医学会学術集会講演要旨集, 2014
  2014 - 2014
• フローサイトメトリーによるプリオン感染細胞の検出
  山崎剛士; 鈴木章夫; 長谷部理絵; 堀内基広
  日本ウイルス学会学術集会プログラム・抄録集, 2014
  2014 - 2014
• プリオンの増殖に関わる細胞内小器官の同定-プリオン感染マウス脳組織の免疫組織学的解析から-
  堀内基広; 斉藤真央; 長谷部理絵; 山崎剛士
  日本ウイルス学会学術集会プログラム・抄録集, 2014
  2014 - 2014
• ウシパイエル板M細胞における解糖系酵素アルドラーゼAを介した異常型プリオン蛋白質取り込み機構
  長澤裕哉; 盛田彰太郎; 日高湧介; 渡邉一史; 渡邊康一; 大和田修一; 北澤春樹; 今村守一; 横山隆; 堀内基広; 坂口末廣; 麻生久
  日本畜産学会大会講演要旨, 2014
  2014 - 2014
• プリオンタンパク質凝集体のQバンド電子スピン二重共鳴法(Q-band DEER)による構造解析
  稲波修; 山盛徹; 安井博宣; 堀内基広; 平岡和佳子; 古川貢; 中村敏和
  電子スピンサイエンス学会年会講演要旨集, 2013
  2013 - 2013
• 補体反応はプリオン感染神経細胞の膜透過性を亢進させる
  長谷部理絵; CAUGHEY Byron; 堀内基広
  日本獣医学会学術集会講演要旨集, 2013
  2013 - 2013
• プリオン感染マウスにおけるミクログリア活性化状態の解析
  堀内基広; 蕪木洋之; 長谷部理絵; 山崎剛士
  日本ウイルス学会学術集会プログラム・抄録集, 2013
  2013 - 2013
• 大脳皮質由来初代神経培養系におけるプリオン感染の解析
  藤原愛; 長谷部理絵; 山崎剛士; 鈴木章夫; 堀内基広
  日本ウイルス学会学術集会プログラム・抄録集, 2013
  2013 - 2013
• マウス腸管M細胞における解糖系酵素アルドラーゼAを介した異常型プリオン蛋白質取り込み機構
  長澤裕哉; 盛田彰太郎; 日高湧介; 渡邉一史; 大和田修一; 北澤春樹; 今村守一; 横山隆; 堀内基広; 坂口末廣; 西田教行; 渡邊康一; 野地智法; 麻生久
  日本獣医学会学術集会講演要旨集, 2013
  2013 - 2013
• プリオン感染初期における異常型プリオンタンパク質の新規産生と細胞内動態の解析
  山崎剛士; 鈴木章夫; 長谷部理絵; 堀内基広
  日本獣医学会学術集会講演要旨集, 2013
  2013 - 2013
• プリオンタンパク質凝集体の電子スピン共鳴-スピンラベル法による解析
  稲波修; 山盛徹; 安井博宣; 堀内基広; 平岡和佳子; 古川貢; 中村敏和
  日本獣医学会学術集会講演要旨集, 2013
  2013 - 2013
• BSE感染モルモットにおける接種ルートによるプリオン分布の相違
  中西勇貴; 西村麻紀; 堀内基広; 古岡秀文
  日本獣医学会学術集会講演要旨集, 2013
  2013 - 2013
• Brucella abortusは腸管パイエル板からの侵入にM細胞上のプリオン蛋白質(PrP^c)を利用する
  鈴木道雄; 中藤学; 中藤学; 渡会雅久; 木村昌伸; 堀内基弘; 長谷耕二; 長谷耕二; 飛梅実; 阿戸学; 森川茂; 山田章雄; 山田章雄; 大野博司; 今岡浩一
  日本獣医学会学術集会講演要旨集, 2013
  2013 - 2013
• 抗原への二価結合により付与されるmAb132のPrP^Sc特異性
  鈴木章夫; 山崎剛士; 長谷部理絵; 堀内基広
  日本ウイルス学会学術集会プログラム・抄録集, 2012
  2012 - 2012
• BSEおよびScrapie由来プリオン経口感染マウスにおける病理学的研究
  藤原理央; 中西勇貴; 堀内基広; 古岡秀文
  日本獣医学会学術集会講演要旨集, 2012
  2012 - 2012
• プリオン感染CD14ノックアウトマウスにおけるミクログリアの活性化とPrP^Sc沈着の解析
  長谷部理絵; 蕪木洋之; 鈴木章夫; 山崎剛士; 堀内基広
  日本ウイルス学会学術集会プログラム・抄録集, 2012
  2012 - 2012
• BSE伝達モルモット小脳におけるプリオン沈着と神経伝達物質に関する病理学的研究
  坂口翔一; 室井喜景; 堀内基広; 古岡秀文
  日本獣医学会学術集会講演要旨集, 2011
  2011 - 2011
• プリオン感染では株により異なる補体因子が反応する
  長谷部理絵; 堀内基広; BYRON Caughey
  日本ウイルス学会学術集会プログラム・抄録集, 2010
  2010 - 2010
• BSE感受性に関わる東南アジアのウシプリオン遺伝子多型
  内田玲麻; HERIYANTO Agus; THONGCHAI Chalermchaikit; HANH Tran Thi; 堀内基広; 石原加奈子; 田村豊
  日本獣医学会学術集会講演要旨集, 2010
  2010 - 2010
• BSE伝達モルモット小脳におけるプリオンの経時的な動態と神経伝達物質への影響
  坂口翔一; 堀内基広; 古岡秀文
  日本獣医学会学術集会講演要旨集, 2010
  2010 - 2010
• 1SA2-02 Intracellular localization of abnormal isoform of prion protein(1SA2 BSJ&ABA Joint Symposium, "Prion and Virus Infections",The 47th Annual Meeting of the Biophysical Society of Japan)
  Horiuchi Motohiro
  Seibutsu Butsuri, 2009, The Biophysical Society of Japan General Incorporated Association, English
  2009 - 2009
• 3P-065 Analysis of Soluble Oligomers of Prion Protein by Fluorescence Correlation Spectroscopy(Protein:Function,The 47th Annual Meeting of the Biophysical Society of Japan)
  Sakata Hiroshi; Horiuchi Motohiro; Kinjo Masataka
  Seibutsu Butsuri, 2009, The Biophysical Society of Japan General Incorporated Association, English
  2009 - 2009
• プリオン蛋白質の可溶性オリゴマーの蛍光相関分光法による解析
  坂田啓司; 堀内基広; 金城政孝
  日本蛋白質科学会年会プログラム・要旨集, 2009
  2009 - 2009
• 抗PrP抗体の末梢投与によるプリオン病の治療効果
  大澤夏生; 宋昌絃; 鈴木章夫; 堀内基広
  日本ウイルス学会学術集会プログラム・抄録集, 2009
  2009 - 2009
• マウスプリオンタンパク質ヒスチジン186周辺の銅イオン結合部位に関する構造解析
  稲波修; 渡辺康子; 五十嵐学; 伊藤公人; 堀内基広; 桑原幹典; 平岡和佳子
  日本獣医学会学術集会講演要旨集, 2009
  2009 - 2009
• Cd14分子のプリオン病病態への関与
  酒井景子; 長谷部理絵; 宋昌紘; 堀内基広
  日本ウイルス学会学術集会プログラム・抄録集, 2009
  2009 - 2009
• 骨髄由来間葉系幹細胞のプリオン感染脳病変への走化に関与する因子の解析
  宋昌鉉; 堀内基広
  日本ウイルス学会学術集会プログラム・抄録集, 2009
  2009 - 2009
• マウス神経芽腫細胞Neuro2aにおけるプリオンの細胞間伝播にはエクソソーム以外の因子が関与する
  堀内基広; 瓜生匡秀; 山崎剛士; 中満智史; 長谷部理絵
  日本獣医学会学術集会講演要旨集, 2008
  2008 - 2008
• PrP^Scのaa81-aa137の領域はプリオンの感染性に必須である
  堀内基広; 長谷部理絵
  日本ウイルス学会学術集会プログラム・抄録集, 2008
  2008 - 2008
• 間接蛍光抗体によるプリオン持続感染細胞に存在するPrP^Scの検出
  山崎剛士; 瓜生匡秀; 中満智史; 堀内基広
  日本ウイルス学会学術集会プログラム・抄録集, 2008
  2008 - 2008
• プリオン感染マウス脳における骨髄由来間葉系幹細胞の動態
  宋昌鉉; 長谷部理絵; 堀内基宏
  日本ウイルス学会学術集会プログラム・抄録集, 2008
  2008 - 2008
• プリオン持続感染マウスNeuro2aサブクローンにおけるPrP^Scの量的変動について
  中満智史; 黒川彩; 瓜生匡秀; 堀内基広
  日本獣医学会学術集会講演要旨集, 2007
  2007 - 2007
• 抗PrP抗体の脳内投与によるプリオン病の治療効果
  宋昌鉉; 古岡秀文; 鈴木章夫; 長谷部理絵; 前田秋彦; 堀内基広
  日本ウイルス学会学術集会プログラム・抄録集, 2007
  2007 - 2007
• 日本の種雄牛におけるBSE感受性に関するプリオン遺伝子多型の出現頻度
  村松康和; 高橋健一; 早川宏之; 堀内基広; 上野弘志; 田村豊
  日本獣医学会学術集会講演要旨集, 2007
  2007 - 2007
• ポリ6-硫酸化N-アセチルグルコサミン誘導体の合成と抗プリオン活性
  永塚健宏; 永塚健宏; 鵜沢浩隆; 堀内基広; 西田芳弘
  日本糖質学会年会要旨集, 2007
  2007 - 2007
• プリオンタンパク質C末端領域の新たなCu²⁺結合構造の同定
  稲波修; 稲波修; 渡邊康子; 渡邊康子; 堀内基広; 堀内基広; 平岡和佳子; 下山雄平; 桑原幹典; 桑原幹典
  日本獣医学会学術集会講演要旨集, 2007
  2007 - 2007
• プリオンタンパク質のpH感受性における塩橋とヒスチジン残基の役割
  渡邊康子; 渡邊康子; 平岡和佳子; 下山雄平; 堀内基広; 堀内基広; 桑原幹典; 桑原幹典; 稲波修; 稲波修
  日本獣医学会学術集会講演要旨集, 2007
  2007 - 2007
• マウス神経芽腫細胞Neuro2a(N2a)サブクローン間におけるプリオンの細胞間伝播の相違
  瓜生匡秀; 堀内基広
  日本獣医学会学術集会講演要旨集, 2007
  2007 - 2007
• プリオンの増殖とその抑制
  堀内基広; 堀内基広
  日本ウイルス学会学術集会プログラム・抄録集, 2007
  2007 - 2007
• プリオンタンパク質β-シート領域のpH感受性における塩橋とヒスチジン残基の役割
  渡邊康子; 渡邊康子; 平岡和佳子; 下山雄平; 堀内基広; 堀内基広; 桑原幹典; 桑原幹典; 稲波修; 稲波修
  生化学, 2007
  2007 - 2007
• 抗PrP抗体の脳室内投与によるプリオン病進行の遅延効果
  宋昌鉉; 古岡秀文; 金チャンラン; 鈴木章夫; 前田秋彦; 堀内基広
  日本獣医学会学術集会講演要旨集, 2007
  2007 - 2007
• 神経芽腫細胞Neuro2aサブクローンにおけるプリオンの細胞間伝播
  瓜生匡秀; 堀内基広
  日本ウイルス学会学術集会プログラム・抄録集, 2007
  2007 - 2007
• プリオン持続感染細胞における異常型プリオン蛋白質の量的変動
  中満智史; 黒川彩; 瓜生匡秀; 堀内基広
  日本ウイルス学会学術集会プログラム・抄録集, 2007
  2007 - 2007
• 日本で発生したBSE由来PrP^Scの生化学性状の解析
  進藤亮; 山河芳夫; 佐多徹太郎; 横山隆; 古岡秀文; 堀内基広
  日本獣医学会学術集会講演要旨集, 2007
  2007 - 2007
• ベトナムの酪農牛におけるプリオン遺伝子の多形性
  村松康和; 要田正治; 小河孝; 堀内基広; 田村豊
  日本獣医学会学術集会講演要旨集, 2006
  2006 - 2006
• プリオン感受性・非感受性Neuro2aサブクローンを用いたプリオン増殖関連宿主因子の探索
  中満智史; 瓜生匡秀; 堀内基広
  日本ウイルス学会学術集会プログラム・抄録集, 2006
  2006 - 2006
• マウス神経芽腫細胞Neuro2a(N2a)サブクローンで検出される異常型プリオン蛋白質(PrP^Sc)の相違
  瓜生匡秀; 堀内基広
  日本ウイルス学会学術集会プログラム・抄録集, 2006
  2006 - 2006
• Site-Directed Spin Labeling(SDSL)法によるプリオンタンパク質pH感受性領域の解析
  渡邊康子; 稲波修; 稲波修; 堀内基広; 下山雄平; 下山雄平; 中村秀夫; 中村秀夫; 桑原幹典
  日本獣医学会学術集会講演要旨集, 2006
  2006 - 2006
• プリオン感受性細胞と非感受性細胞の遺伝子発現比較解析
  中満智史; 瓜生匡秀; 堀内基広
  日本膜学会年会講演要旨集, 2006
  2006 - 2006
• プリオンタンパク質の抗酸化機能の解明
  渡邊康子; 渡邊康子; 稲波修; 稲波修; 井上哲; 飯塚大輔; 堀内基広; 堀内基広; 桑原幹典
  電子スピンサイエンス学会年会講演要旨集, 2006
  2006 - 2006
• 細胞におけるプリオンの増殖
  堀内基広
  日本膜学会年会講演要旨集, 2006
  2006 - 2006
• 動物プリオン病の病態形成に関与するサイトカインの探索
  古岡秀文; 薮添敦史; 田村勇耕; 堀内基広; 佐多徹太郎
  日本獣医学会学術集会講演要旨集, 2006
  2006 - 2006
• BSE問題とプリオン病研究の現状
  堀内基広
  BMSコンファレンス講演要旨集, 2005
  2005 - 2005
• プリオン感染動物におけるAHSP mRNA量低下の分子機構:造血系細胞に対する直接作用の検証
  勝岡加代子; 大塚弥生; 大塚弥生; 伊東大介; 伊東大介; 大田寛; 大田寛; 堀内基広; 堀内基広; 稲葉睦; 稲葉睦
  日本獣医学会学術集会講演要旨集, 2005
  2005 - 2005
• プリオンタンパク質の銅イオンの新たな結合様式
  稲波修; 稲波修; 堀内基広; 堀内基広; 平岡和佳子; 稲垣冬彦; 下山雄平; 下山雄平; 中村秀夫; 中村秀夫; 桑原幹典
  日本獣医学会学術集会講演要旨集, 2005
  2005 - 2005
• プリオンタンパク質過剰発現細胞における酸化ストレス応答
  井上哲; 稲波修; 飯塚大輔; 堀内基広; 桑原幹典
  日本獣医学会学術集会講演要旨集, 2005
  2005 - 2005
• モンゴル在来ヒツジからのピロプラズマ目原虫DNAの検出
  村松康和; 八幡陽介; 堀内基広; 石黒直隆; 品川森一; 森田千春; 田村豊
  日本獣医学会学術集会講演要旨集, 2005
  2005 - 2005
• マウスプリオンタンパク質α-ヘリックス領域のSite-Directed Spin Labeling(SDSL)法を用いた解析
  渡辺康子; 渡辺康子; 稲波修; 稲波修; 稲波修; 堀内基広; 堀内基広; 下山雄平; 下山雄平; 中村秀夫; 中村秀夫; 桑原幹典
  日本獣医学会学術集会講演要旨集, 2005
  2005 - 2005
• 蛍光相関分光法による未変性条件下での異常型プリオン蛋白質の検出
  堀内基弘; 品川森一
  日本ウイルス学会学術集会プログラム・抄録集, 2005
  2005 - 2005
• マウス神経芽腫細胞Neuro-2aのプリオン感受性はPrP^C以外の因子により規定される
  瓜生匡秀; 堀内基広
  日本ウイルス学会学術集会プログラム・抄録集, 2005
  2005 - 2005
• 現行の牛海綿状脳症検査のシカ慢性消耗病(CWD)診断への応用性
  嶋田希実; 岩丸祥史; 林浩子; 高田益宏; 牛木祐子; 木村久美子; 田川裕一; 堀内基広; 品川森一
  日本獣医学会学術集会講演要旨集, 2004
  2004 - 2004
• Inhibitory Activity of Sulfonated GlcNAc Derivatives against Sialidases and Scrapie Prions
  佐々木健二; 西田芳弘; 神原実季恵; 堀内基広; 鵜沢浩隆; 小林一清
  日本化学会講演予稿集, 2004
  2004 - 2004
• プリオンの部位特異的スピンラベル法(SDSL-ESR)による構造変化の解析
  稲波修; 橋田修吉; 飯塚大輔; 井上哲; 堀内基広; 桑原幹典
  日本獣医学会学術集会講演要旨集, 2004
  2004 - 2004
• マウススクレイピーモデルを用いたプリオン蛋白質に対する宿主免疫応答の再考
  片岡那津見; 石黒直隆; 前田秋彦; 堀内基広
  日本獣医学会学術集会講演要旨集, 2004
  2004 - 2004
• ウシプリオン遺伝子の多型解析
  中満智史; 石黒直隆; 前田秋彦; 堀内基広; 宮沢孝幸
  日本獣医学会学術集会講演要旨集, 2004
  2004 - 2004
• Site-directed spin label(SDSL)法によるマウスプリオンタンパク質の銅イオン結合部位周辺の構造解析
  稲波修; 橋田修吉; 飯塚大輔; 堀内基広; 平岡和佳子; 稲垣冬彦; 桑原幹典
  電子スピンサイエンス学会年会講演要旨集, 2004
  2004 - 2004
• 6位硫酸化N-アセチルグルコサミンクラスターの異常プリオン産生阻害活性
  神原実季恵; 西田芳弘; 佐々木健二; 篠田康彦; 小林一清; 堀内基広; 鵜沢浩隆
  高分子学会予稿集(CD-ROM), 2004
  2004 - 2004
• 「BSEの解明と対策の最前線」プリオン蛋白質とプリオン病
  堀内基広
  日本畜産学会大会講演要旨, 2004
  2004 - 2004
• 人獣共通感染症としてのプリオン病
  堀内基広
  日本ウイルス学会学術集会プログラム・抄録集, 2004
  2004 - 2004
• スクレイピー持続感染細胞I3/I5におけるプリオン増殖
  荻野倫子; 菊池宏明; 前田秋彦; 堀内基広
  日本ウイルス学会学術集会プログラム・抄録集, 2004
  2004 - 2004
• 新生子マウスの腸粘膜上皮細胞へのプリオン伝達の試み
  岡本実; 古岡秀文; 堀内基弘; 野口敬紀; 萩原克郎; 村松康和; 辻正義; 朝長啓造; 生田和良
  日本獣医学会学術集会講演要旨集, 2003
  2003 - 2003
• プリオンの環境汚染評価-植物への影響
  三隅智子; 石黒直隆; 堀内基広; 品川森一
  日本獣医学会学術集会講演要旨集, 2003
  2003 - 2003
• 経口ルートによるプリオンの感染成立には消化管リンパ装置の存在が必要である
  堀内基広; 石黒直隆; 品川森一; 古岡秀文; 北村延夫
  日本獣医学会学術集会講演要旨集, 2002
  2002 - 2002
• エポキシ化合物によるプリオン不活化
  山本真理; 品川森一; 石黒直隆; 堀内基広
  日本獣医学会学術集会講演要旨集, 2001
  2001 - 2001
• in vitro試験系による病原性プリオン蛋白質と正常プリオン蛋白質の相互作用の解析
  堀内基広
  日本獣医学会学術集会講演要旨集, 2001
  2001 - 2001
• プリオン蛋白質とプリオン病
  堀内基広
  タンパク質構造討論会講演要旨集, 2000
  2000 - 2000
• Distribution of scrapie prion protein in sheep body. Immunohistochemical retrieval by anti-PrP antibody.
  鈴木智; 高橋広志; 松井高峯; 堀内基広; 石黒直隆; 品川森一; 古林与志安; 古岡秀文
  日本獣医学会学術集会講演要旨集, 1998
  1998 - 1998
• Sheep biodistribution of scrapie prion protein. PrPSc detection by Western blotting.
  高橋未緒; 堀内基広; 石黒直隆; 松井高峯; 古岡秀文; 品川森一
  日本獣医学会学術集会講演要旨集, 1998
  1998 - 1998
• Analysis of cattle prion protein PrP gene promoter region.
  井上真吾; 田中成幸; 堀内基広; 石黒直隆; 品川森一
  日本獣医学会学術集会講演要旨集, 1996
  1996 - 1996

• アドバンスト演習, 2024年, 学士課程, 獣医学部
• 大学院共通授業科目（一般科目）：複合領域, 2024年, 修士課程, 大学院共通科目
• 感染症学特別研究Ⅰ, 2024年, 博士後期課程, 国際感染症学院
• 大学院共通授業科目（一般科目）：複合領域, 2024年, 修士課程, 大学院共通科目
• 感染症学特別演習, 2024年, 博士後期課程, 国際感染症学院
• 研究機器演習（総合成績）, 2024年, 博士後期課程, 獣医学院
• 感染症学特別研究ⅡA, 2024年, 博士後期課程, 国際感染症学院
• 研究機器演習（総合成績）, 2024年, 博士後期課程, 国際感染症学院
• 感染症学特別研究ⅡB, 2024年, 博士後期課程, 国際感染症学院
• 海外インターンシップA, 2024年, 博士後期課程, 国際感染症学院
• 研究機器演習, 2024年, 博士後期課程, 国際感染症学院
• 海外インターンシップB, 2024年, 博士後期課程, 国際感染症学院
• 研究機器演習, 2024年, 博士後期課程, 獣医学院
• 獣医衛生学特論, 2024年, 博士後期課程, 獣医学院
• 獣医衛生学特論, 2024年, 博士後期課程, 国際感染症学院
• 人獣共通感染症対策専門特論, 2024年, 博士後期課程, 国際感染症学院
• アカデミックイングリッシュ, 2024年, 博士後期課程, 獣医学院
• アカデミックイングリッシュ, 2024年, 博士後期課程, 国際感染症学院
• 応用動物衛生学, 2024年, 学士課程, 獣医学部
• 獣医英語演習, 2024年, 学士課程, 獣医学部
• 研究スタートアップ演習, 2024年, 学士課程, 獣医学部
• 食品衛生学実習, 2024年, 学士課程, 獣医学部
• 微生物学, 2024年, 学士課程, 医学部
• 基礎動物衛生学, 2024年, 学士課程, 獣医学部
• アドバンスト演習, 2024年, 学士課程, 獣医学部
• 食品衛生学, 2024年, 学士課程, 獣医学部
• アドバンスト演習, 2024年, 学士課程, 獣医学部
• アドバンスト演習, 2024年, 学士課程, 獣医学部
• アドバンスト演習, 2024年, 学士課程, 獣医学部
• アドバンスト演習, 2024年, 学士課程, 獣医学部

• Identification of neuronal cell-type vulnerable to prion infection and analysis of molecular mechanism of the induced neuronal cell death
  Grants-in-Aid for Scientific Research
  01 Apr. 2025 - 31 Mar. 2028
  堀内 基広
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 25K02167
• ワンヘルスアプローチによるモンゴルにおける食中毒起因菌と薬剤耐性菌の調査研究
  科学研究費助成事業
  Sep. 2024 - Mar. 2028
  堀内 基広; 佐藤 豊孝; 笹岡 一慶
  日本学術振興会, 国際共同研究加速基金(海外連携研究), 北海道大学, ［Internationally co-authored］, 24KK0134
• Studies on the control of disease progression of prion diseases by transition of glial cell polarization
  Grants-in-Aid for Scientific Research
  01 Apr. 2022 - 31 Mar. 2025
  堀内 基広
  本研究では、緻密骨由来間葉系幹細胞 (CB-MSCs) がアストロサイト (AS) およびミクログリア (MG) の極性転換に関与する因子の同定、および、極性が転換したグリア細胞の神経保護・傷害機能の解析を行い、グリア細胞の極性転換法を、プリオン病を含む難治性神経変性疾患の病態進行を制御する方法の開発へ応用することを目的とする。
  本年は、ASにプリオンが感染・増殖した場合に、神経細胞傷害性のA1型に極性がシフトするか、および、小胞体ストレス応答が増大するかを詳細に検討した。初代ASにプリオンを添加して、プリオンの感染効率、細胞内局在、および増殖を調べたところ、PrPScが細胞膜上に繊維状に存在する初代神経細胞と異なり、ASでは、核周囲に顆粒状にPrPScが存在した。このPrPScはＮ末端側のプロセッシングを受けていること、および、Neuro2a神経芽細胞におけるPrPScの細胞内局在と同様の局在を示すことから、主に、細胞内膜輸送経路上に存在すると考えられた。50-70%程度のASが、核周囲の明瞭なPrPSc陽性像を示すにもかかわらず、A1型への極性のシフト、および小胞体ストレス応答の増加を示す結果は得られなかった。ASにけるPrPScのの蓄積が経時的に著しく増加する傾向は認められず、約3週間の培養で、2倍弱の増加であったことからも、PrPScの増殖単独では、ASに大きな極性変化をもたらさないと結論した。現在、プリオン感染ミクログリアで、プリオン感染/非感染ASを刺激して、感染/非感染によるASの応答を解析中である。
  CB-MSCs のMGの極性転換に関与する可能性のある分子として、プリオン感染局所（組織）とプリオン感染MGのRNA sequenceの結果から、EGF1を見いだしたが、抗EGF1抗体はMGの極性転換を阻害しなかった。
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 23K23772
• Studies on the control of disease progression of prion diseases by transition of glial cell polarization
  Grants-in-Aid for Scientific Research
  01 Apr. 2022 - 31 Mar. 2025
  堀内 基広
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 22H02507
• 非定型ＢＳＥ等動物プリオン病のヒトへの感染リスクの推定と低減に資する研究
  厚生労働科学研究費補助金
  Apr. 2020 - Mar. 2023
  堀内基広; 古岡秀文; 新竜一郎; 宮澤光太郎; 飛梅実; 萩原健一; 小野文子
  厚生労働省, 食品の安全確保推進研究事業, 北海道大学大学院獣医学研究院
• Role of stress-induced transcription factor ATF3 on neurodegeneration of prion diseases
  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
  01 Apr. 2019 - 31 Mar. 2022
  HORIUCHI MOTOHIRO
  In this study, we analyzed the role of stress-inducible activating transcription factor 3 (ATF3) in the pathobiology of prion diseases. ATF3-positive cells were significantly increased in the thalamus and pontine nucleus of prion-infected mice, where neuronal cell death is well-observed, and around 80% of ATF-3 positive cells were identified as neurons. Apoptosis were not observed in the thalamus, where pronounced neuronal cell death occurred. The amount of GPx4 in the thalamus tended to decrease with prion infection. The 4-HNE, a marker of lipid peroxidation, was less accumulated in ATF3-positive neurons than ATF3-negative neurons. These results suggested that neurons in the thalamus die of ferroptosis and that induction of ATF3 expression may mitigate lipid peroxidation in the neurons and suppress ferroptosis.
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 19H03119
• 食品を介する家畜・家禽疾病のリスク管理に関する研究
  厚生労働科学研究費補助金
  Apr. 2017 - Mar. 2020
  新 竜一郎; 柴田 宏昭; 萩原 健一; 飛梅 実; 古岡 秀文; 松浦 裕一; 山﨑 剛士; 鎌田 洋一; 壁谷 英則; 森田 幸雄; 保富 康宏
  厚生労働省, 食品の安全確保推進研究事業, 北海道大学, Principal investigator
• Elucidation of the mechanism for neurodegeneration in prion diseases through neuron-glia cross-talk
  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A)
  01 Apr. 2015 - 31 Mar. 2019
  HOROUICHI MOTOHIRO; KURODA minori; SHAN zhifu
  Microglia activation and astrogliosis are observed in the early stage of prion diseases. Therefore, glial activation are thought to be involved in the neurodegeneration, however, details of the mechanisms remain to be elucidated. In this study, first we analyzed the activation state of microgila and astrocyte in prion infected mice by RNA-sequencing. We also analyzed brain regions where neuronal loss was progressively observed in prion-infected mice and analyzed gene expression in the small region. Neuron-enriched genes were selected by bioinformatics and further analyses disclosed that expression of stress-induced transcriptional factor, ATF3, was induced in the lateral dorsal part of the thalamus where progressive neuronal loss was observed. The lateral dorsal part of the thalamus is one of the regions where intense glial activation is observed and thus, it is of interest to elucidate if glial activation induces STF3 induction in prion-infected neurons.
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (A), Hokkaido University, 15H02475
• Identification of MSC subpopulations that have therapeutic potential for neurodegenerative disorders
  Grants-in-Aid for Scientific Research
  01 Apr. 2016 - 31 Mar. 2018
  HORIUCHI MOTOHIRO
  Intra-cerebral transplantation of compact bone-derived MSCs (CB-MSCs) prolonged survival of prion-infected mice, suggesting that CB-MSCs could be useful for the treatment of neurodegenerative disorders. In the CB-MSCs transplanted prion-infected mice, unexpectedly, microglia were more activated than un-transplanted mice. However, up-regulation of some of the marker genes for M2-type microglia, such as Arg-1, suggesting that CB-MSCs modify the activation state of microglia. The culture supernatant of CB-MSCs also shifted the activation state of microglia recovered from prion-infected mice to M2-type activation state. These results suggest that the alteration of innate immunity by CB-MSCs is one of the mechanisms for the therapeutic effects of CB-MSCs on neurodegenerative disorders.
  Japan Society for the Promotion of Science, Grant-in-Aid for Challenging Exploratory Research, Hokkaido University, 16K15033
• Role of functional change of microglia in neuropathogenesis of prion diseases
  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A)
  01 Apr. 2011 - 31 Mar. 2015
  HORIUCHI Motohiro; INANAMI Osamu; HASEBE Rie; SASSA Yukiko; OHSIMA Masanobu
  We established PrPSc-specific staining method and using this method we elucidated a part of neuropathophysiology in the early stage of prion infection; astrocytes recognize prion propagation directly or subtle changes in neurons by prion propagation and astrocyte activation precedes over microglial activation.
  Gene expression analysis of microglia in prion-infected mice showed that microglia play a neuroprotective role by secreting neurotrophic factors at the early stage of prion infection in the early stage of prion infection, but the microglial activation state shifts to pro-inflammatory or neurotoxic state along with the progression of prion disease. Furthermore, anti-inflammatory cytokines such as IL-10 interfere the propagation of prions in brain.
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (A), Hokkaido University, Principal investigator, Competitive research funding, 23248050
• Identification of factors that involved in migration of mesenchymal stem cells to lesions of neurodegenerative diseases
  Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research
  2011 - 2012
  HORIUCHI Motohiro
  In vitro migration assay was carried out to investigate chemoattractive factors for MSCs in the brain lesions of prion-infected mice. In addition, expression of chemokine receptor on MSCs that had been transplanted in the brain of prion-infected mice was analyzed. The combined in vitro and in vivo analyses suggest that CCR3, CCR5, CXCR3 and CXCR4 and their corresponding ligands are involved in the migration of hMSCs to the brain lesions caused by prion propagation.
  Japan Society for the Promotion of Science, Grant-in-Aid for Challenging Exploratory Research, Hokkaido University, Principal investigator, Competitive research funding, 23658233
• Structural analysis of zoonosis-related high-molecular protein aggregation by double electron-electron resonance (DEER) technique
  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
  2009 - 2011
  INANAMI Osamu; INABA Mustumi; HORIUCHI Motohiro; KUWABARA Mikinori; YAMAMORI Tohru
  In this study, to obtain the structural information for this pathological fibrous form of PrPs', we measured the intermolecular distance in the pathological fibrous form with D178N mutation by the site-directed spin labeling (SDSL) with continuous wave (CW) ESR (X-band) and pulsed ESR (Q-band). After the formation of the fibrous structure by exposure to 1M of guanidine hydrochloride in pH4.0, the ESR spectra were measured by CW-ESR. The interspin-distances in pathological fibrous form were measured by continuous wave ESR (CW-ESR) and the four-pulse double electron-electron resonance (DEER) technique. The information of intermolecular distances will be useful to estimate the pathogenic fibrous structure of the familial prion disease, since it is difficult to analyze the macromolecules such as the fibrous protein by NMR or X-ray crystallography.
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, Coinvestigator not use grants, Competitive research funding, 21380185
• 再生医療を応用したプリオン病治療モデルの確立
  科学研究費助成事業 挑戦的萌芽研究
  2008 - 2009
  堀内 基広; 宋 昌鉉; 長谷部 理絵
  プリオン病治療法の確立を目的として、骨髄由来間葉系幹細胞(Bone marrow-derived Mesenchymal stem cells, MSC)のプリオン病の治療への応用について、プリオン感染マウスをモデルとして検討した。ヒト不死化MSCs(hMSCs)を、Chandler株感染マウスが初期の臨床症状を呈する接種後120日に海馬に移植した場合、非移植対照群(150±2日)と比べて潜伏期が有意に延長した(157±6日)。また、hMSCsを尾静脈から移植した場合、非移植対照群(148±6日)と比べて潜伏期が有意に延長した(158±6日)。hMSCsを移植したマウスでは、PrP^の蓄積量は対照群と比較して差は認められなかったが、空胞変性は軽度であった。従って、hMSCsはプリオンの増殖は阻害しないが、病気の進行に抑制的に働くことが明らかとなった。hMSCsはプリオン感染マウス脳内で、神経細胞様、アストロサイト様、およびオリゴデンドロサイト様の細胞に分化したが、その効率は低いことから、hMSCsの移植による潜伏期の延長は、bystander effectによるものと考えられた。
  hMSCsはいずれのルートで移植した場合でもプリオン病の神経病変へ移行したことから、hMSCsがプリオン病の神経病変部に移行するメカニズムを調べるために、in vitro走化試験をスクリーニング系として用いて、サイトカイン、ケモカインおよび栄養因子およびそれらのレセプターの関与を調べた。その結果、CCR3,CCR4,CCR4およびCXCR4のケモカインレセプターとそのリガンドがhMSCsの走化に関与していることが示唆された。プリオン感染マウスの視床に移植したhMSCsは脳梁を通って、反対側の海馬へと移動するが、移植後、脳梁に存在するhMSCsではCCR3,CCR4,CCR4およびCXCR4のケモカインレセプターが発現していたことから、実際の神経組織においてもCCR3,CCR4,CCR4およびCXCR4およびそのリガンドは、hMSCsのプリオン病病変部の走化に関与することが明らかとなった。
  日本学術振興会, 挑戦的萌芽研究, 北海道大学, Principal investigator, Competitive research funding, 20658070
• Elucidation of the mechanism on the formation of early pathological lesion of prion diseases
  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A)
  2006 - 2009
  HORIUCHI Motohiro; INANAMI Osamu; KIMURA Kazuhiro; OCHIAI Kenji; HASEBE Rie; URYU Masahide; SON Chang-hyun; OOSHIMA Masanobu; FURUOKA Hidefumi
  To identify host genes whose expression is up-regulated in the early stage of prion-infection, DNA microarray analysis was carried out and involvement of the genes in pathobiology of prion diseases was analyzed. The results showed that Cd14 molecule, known as receptor for LPS, influences the progression of the disease, whereas, Cxcl10, known as chemokine with neuroprotective effect, plays a role in antagonizing the disease progression. In addition, microglial activation was suggested to have a protective effect on the disease progression.
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (A), Hokkaido University, Principal investigator, Competitive research funding, 18208026
• Molecular imaging of prion protein by atomic force microscop yand a novel electro-spin-resonance techniques
  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
  2007 - 2008
  I NANAML Osamu; INABA Mutsumi; HORIUTCHI Motohiro; KUWABARA Mikinori; ASANUMA Katetoshi; HIRAOKA Wakako; SHINOYAMA Yuhei
  本研究はブリオン病の原因であるブリオンタンパク質の凝集体変化を新しい方法を用いて明らかにし、今後のプリオン病の病態解明や治療薬の評価方法を開発することを目的とした。このプリオン凝集体への変化を起こすには細胞内低pH器官であるエンドソームで起きることから、pHの低下が必須であると考えられていることからpH7.0の生理条件からpH4.0に変化させたときの構造変化を明らかにした。電子スピン共鳴法、ならびに原子間力顕微鏡を用いて家族性プリオン病でよく見られる変異体プリオンD177NはpH4.0、変性下に置くことで変異を持っていない野生型よりも効率よく線維状の凝集体が起きることが示され。凝集の引き金となる領域がタンパク中心部のβシート付近にあることが明らかとなった。今回の結果はプリオン病の原因タンパク質の構造変化を明らかにしただけでなく、今回用いた新たな方法はプリオン関連病の治療薬の評価系に用いることが可能であると考えられる。
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, Coinvestigator not use grants, Competitive research funding, 19380172
• STUDY ON THE NERVE-IMMUNE SYSTEMS AND CELLULAR FACTORS INFLUENCE TO THE PATHOGENICTTY AND DISTRIBUTION OF PRP^IN ANIMAL PRION DISEASES
  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
  2005 - 2007
  ISHIGURO Naotaka; YANAI Takuma; KUWATA Kazuo; HORIUCHI Motohiro
  The pathogenesis of animal prion diseases is influenced by prion strain, genetic factors, natural immunology and several cellular factors in the affected hosts. In this study, genetic factors, nerve-immune system and uptake of PrP^ by macrophages were mainly studied. The following results were obtained.
  1. Polymorpbisms of PrP gene: Polymorphisms of the 863 cattle, 118 caprine and 136 sika dear (Cervus Nippon) PrP genes in Japan were examined. Several single-nucleotide polymorphisms (SNPs) were found in cattle and sika dear, but not find any amino acid substitutions. Two polymorphisms associated with the resistance to scrapie were found in 118 caprine.
  2. Bovine and sheep nerve systems: To determine the location, namber and segmental distribution of sympathetic, parasympathetic and sensory neurons innervating the ileum in cattle and sheep, tracer experiments with horseradish peroxidase were conducted. The labeled neurons were detected in the celiac and cranial mesenteric ganglion complex in both cattle and sheep. Only a few labeled neurons existed in the caudal mesenteric ganglion and the paravertebral ganglia.
  3. Uptake PrP^ by immune cells: To clarify the uptake the PrP^ by immune cells, the involvement of bovine macrophage in the early stage of prion infection was studied. Brain homogenates of scrapie and BSE were degraded sequentially in bovine macrophage and bovine peripheral immune cells.
  A part of this research has been published in the papers, the other parts are now prepared in the papers.
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Gifu University, Coinvestigator not use grants, Competitive research funding, 17380180
• プリオン感染マウスモデルを用いたプリオン病治療候補薬の評価
  科学研究費助成事業 特別研究員奨励費
  2005 - 2006
  堀内 基広; KIM C.-I.; KIM C.-l
  培養細胞レベルでPrP^の産生を抑制する物質は、プリオン病治療薬の候補となりうる。そこで、候補薬物をプリオン感染モデルで治療効果や副作用を評価して、プリオン病治療法の開発に役立てることを目標とした。プリオン病治療薬は中枢神経系組織に到達することが重要であるので、候補物質を浸透圧ポンプを用いて脳室内に直接投与して副作用、延命効果、治療効果の評価を実施した。候補物質の一つである人工合成硫酸化糖(4SGN,p6SGN)を、スクレイピー帯広株を接種して発病初期にあるマウスに浸透圧ポンプを用いて一ヶ月間脳室内に投与したが、陰性対照と比較して有意な延命効果は認められなかった。特定の抗PrP抗体が培養細胞においてプリオンの増殖を阻害することが知られている。そこで、プリオン増殖阻害活性を有する抗体を脳室内に投与した時の副作用を評価した。用いた3種類の抗PrP抗体のうち、1種類の抗体が、海馬錐体細胞を選択的に変性させることが判明したが、他の2種の抗体は顕著な神経変性を惹起しなかった。抗体が認識するエピトープにより神経変性誘発が異なったことから、神経変性作用のある抗体を培養細胞レベルでスクリーニングして排除することを日的に、マウス神経芽腫細胞(N2a)を抗体で刺激してDNAマイクロアレイ法により、抗体刺激により変動する遺伝子群の解析を実施した。しかし、細胞死に関連する遺伝子群が特異的に変化しているという現象は認められなかった。現在、神経毒性を誘発しない抗PrP抗体を脳室内に持続投与し、治療効果の評価を継続中である。
  日本学術振興会, 特別研究員奨励費, 北海道大学, 05F05212
• Structural analysis of prion proteins and mechanism of PrPsc transition by using a novel dynamic molecular structure analysis
  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
  2005 - 2006
  INANAMI Osamu; HORIUCHI Motohiro; KARIWA Hiroaki; INABA Mustumi; KUWABARA Mikinori
  We examined the influence of D177N (178N in humans) mutation on the conformational stability of the S2 region of moPrPc with varying pHs by using the SDSL-ESR technique. We prepared moPrPc mutants that reacted with methane thiosulfonate spin-probes (Y161R1 and Y161R1/D177N). The ESR spectrum of D177N at pH 7.5 was narrower than that of Y161R1, referred to as WT^*. The ESR spectrum of D177N did not change when pH in the solution decreased from 7.5 to 4.0.These results suggested that the disappearance of a salt bridge (D177-R163) induced the increase in the instability of S2 region. The values of 1/ H of the central component (Mi=0) in the ESR spectrum obtained from WT^* remained constant from pH 7.5 to pH 6.5, whereas an abrupt increase of 1/Ho occurred when the pH in the solution decreased to under 6.0. These findings indicated that the conformational transition from a rigid structure to a flexible structure existed at between pH 6.5 and pH 6.0. Moreover, the line shape of the ESR spectrum obtained from H176S neighboring the salt bridge linked to the S2 region was narrower than that of WT^* at pH 7.5. When the pH in the solution decreased from 7.5 to 4.0, the change in the spectrum of H176S was small. These results indicate that the protonation of H 176 is strongly associated with the stability of S2 region. These findings are important for understanding the mechanism by which the disruption of the salt bridge in the S2 region forms the pathogenic PrP_Sc structure in hereditary Creutzfeldt-Jacob disease and fatal familial insomnia.
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, Coinvestigator not use grants, Competitive research funding, 17380178
• Molecular pathology for down-regulation of erythroid-specific genes in prion diseases
  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A)
  2004 - 2006
  INABA Mutsumi; HORIUCHI Motohiro; INANAMI Osamu; UMEMURA Takashi
  AHSP is an erythroid-specific molecular chaperone that stabilizes newly synthesised a-globin. Previous studies demonstrated that mRNA levels of AHSP were specifically reduced in hematopoietic tissues of prion-infected animals. The purpose of the present study is to clarify the mechanism for down-regulation of AHSP transcription. A series of truncation and mutation analyses on 2.5-kb 5' upstream region of the AHSP gene in MELhide8 cells and electrophoretic mobility shift assay showed that the minimal 5'-promoter region located at-328-286 to the translation initiation site including a GATA binding motif. Either of two upstream GATA elements at-403 and-381 enhanced reporter gene transcription only in the presence of the minimal GATA element described above. These findings indicate that an erythroid-specific transcription factor GATA-1 is essential to AHSP gene expression and suggest that the down-regulation of AHSP involves changes in GATA-1 transcriptional activation. There was no significant change in gene expression of AHSP, a-globin, b-globin, GATA-1, EKLF, and NF-E2 in MELhide8 cells when the cells were incubated with brain homogenates from scrapie-infected mice for up to 120 hours. Moreover, MELhide8 cells exhibited no accumulation of PrPsc even after 16 passages. These data demonstrated that Prrc has no direct effect on AHSP gene expression in erythroid cells. Instead, IL-6 significantly and IL-1β weakly reduced the expression of AHSP mRNA levels and the AHSP promoter-reporter gene expression in MELhide8 cells in a dose-dependent manner. The reduction was recovered in the presence of the inhibitor of the STAT3 pathway, suggesting that the signal transduction of an inflammatory cytokine IL-6 through STAT3 pathway would modulate GATA-1/AHSP promoter interaction and subsequently causes down-regulation of the AHSP gene.
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (A), Hokkaido University, Coinvestigator not use grants, Competitive research funding, 16208030
• Module Assembly and Evaluation of Artificial Glycosaminoglycans
  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
  2004 - 2006
  NISHIDA Yoshihiro; MIURA Yoshiko; HORIUCHI Motohiro; UZAWA Hirotaka
  Sulfated glycosaminoglycans (GAGs) and their analogues such as pentosan polysulfate (PPS), dextran sulfate (DS), and heparin inhibit abnormal isoform of prion protein (PrP^) formation in prion-infected cells and prolong the incubation time of scrapie-infected animals. Sulfation of GAGs is not completely regulated and possible sulfation sites are randomly sulfated. This property impedes an elucidation of fundamental structures of GAGs that are involved in diverse biological events on cell surfaces. To address the structure-activity relationship of GAGs in the inhibition of PrP^ formation, we assembled a series of glycosaminoglycans analogues by applying our synthetic strategy based on the concept of carbohydrate modules. Among the synthetic glycosides and their copolymers thus derived and examined, monomeric 4-sulfo-N-acetyl-glucosamine (4SGN), and two copolymers, poly-4SGN and poly-6-sulfo-N-acetyl-glucosamine (6SGN), inhibited PrP^ formation with 50% effective dose lower than 20 μg/ml. The inhibitory effect became more evident in consecutive treatment. They reduced the expression of cellular prion protein but did not affect cell growth. Structural comparison suggested that coincidence of N-acetyl group at C-2 with sulfate group at C-4 or C-6 might be involved in the inhibition of PrP^ formation. However, neither monomeric nor dimeric 6SGN, but poly-6SGN showed the inhibitory effect, suggesting the importance of polyvalent configuration in the effect of 6SGN. These results indicate that the artificially sulfated glycosides and their polymers are useful not only for the analysis of structure-activity relationship of GAGs but also for the improvement of new therapeutic compounds for prion diseases.
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), 名古屋大学->千葉大学, Coinvestigator not use grants, Competitive research funding, 16350063
• Is the wild deer population considered to be the source of infectious diseases for other animals and human being?
  Grants-in-Aid for Scientific Research(基盤研究(B))
  2004 - 2006
  Masatsugu SUZUKI; 梶 光一; 山内 貴義; 有川 二郎; 横山 真弓; 堀内 基広; 木村 享史
  1. Nine hundred and seventy six sika deer serum samples were examined. Although 25 (2.6%) of them were positive for anti-HEV IgG, the antibody, titers were very low. HEV RNA was not detected in these samples.2. In 87 wild boar, 2.6% of them were positive for anti-HEV IgG, the antibody. HEV RNA was detected in 2.3% in these samples.3. Twelve serum samples (13.6%) from88wild boar were positive for anti-leptosira IgG.4. By using LAMP and RT-PCR methods, M. paratuberculosis DNA was not detected in 173 samples of wild deer.5. In wild deer blood samples collected in Hokkaido, DNA of Rickettsia He...
  Ministry of Education, Culture, Sports, Science and Technology, 基盤研究(B), 北海道大学->岐阜大学, Coinvestigator not use grants, Competitive research funding, 16380202
• 硫酸化糖を分子基盤とする抗BSE試薬の開発と異常プリオン抑制機構の解析
  科学研究費助成事業 萌芽研究
  2004 - 2005
  西田 芳弘; 三浦 佳子; 小川 智久; 堀内 基広; 小林 一清
  硫酸化ペントサン、ヘパリンなどのグリコサミノグリカンや硫酸多糖体がプリオンの増殖阻害活性を有することは古くから知られている(Ehlers & Diringer, J Gen Virol.,65:1325-1330,1984. Caughey & Raymond, J Virol.,67:643-650,1993.)。これらは、長年にわたり、プリオン病治療法への応用が期待されてきたが、現在まで、硫酸多糖体を使用したプリオン病治療法は確立されていない。天然の硫酸多糖体は硫酸化やアセチル化の部位や割合などが制御されていないため、硫酸多糖体の構造とプリオン増殖阻害活性の構造活性相関の解析が困難である。プリオン増殖阻害活性を担う硫酸化糖の基本骨格が明らかになれば、これをリード化合物として、新たなプリオン病治療薬が創出できる可能性がある。そこで、本研究では、硫酸化部位を正確に制御した合成硫酸化糖を研究代表者の糖鎖モジュール化法を適用することで合成を行い、硫酸化糖の構造とプリオン増殖阻害活性の関係について解析することを目的とした。
  (1)合成硫酸化糖によるプリオン増殖阻害試験
  プリオン持続感染細胞I3/I5-9は、10%FBSを含むOpti-MEM(Invitorgen)で培養した。硫酸化糖はMili-Qに溶解して、使用直前に0.45μmのフィルターで濾過滅菌した。硫酸化糖を加え2日間培養した後、PrP^検出用の試料を調整した。陽性対象として、プリオン増殖阻害活性を有する硫酸デキストラン500(DS)を使用した。PrP^の検出用試料の調整は2-2(5)に述べた通りに行った。また、SDS-PAGEおよびウエスタンブロットは共同研究者の堀内らの方法に従って実施した。
  (2)結果・成果
  硫酸化糖のプリオン増殖阻害活性をPrP^の産生量を指標に評価した。8種の合成硫酸化糖のPrP^産生阻害活性を比べた。陽性対象であるDS500(DS)は1μg/mlの濃度でも、PrP^の産生を70%程度阻害した。調べた硫酸化糖のうち、Hは濃度依存的にPrP^産生を阻害した。検量線グラフからPrP^量を陰性対照の50%にする50%効果濃度(ED_<50>)は約20μg/mlと推定された。また、硫酸化糖Bも100μg/mlでPrP^量が陰性対照の50%以下となり、弱いながらもPrP^産生阻害活性を有する可能性が示唆された。
  日本学術振興会, 萌芽研究, 名古屋大学, Coinvestigator not use grants, Competitive research funding, 16651107
• Molecluar mechanism of propagation of abnormal prion protein in the cells
  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A)
  2003 - 2005
  HORIUCHI Motohiro; INABA Mutsumi; OHASHI Kazuhiko; MAEDA Akihiko; FURUOKA Hidefumi
  In this research, we studied on the molecular mechanism of prion propagation in the cells through analyses of the effect of compounds that inhibit PrP^ formation and identification of host factor(s) and microenvironment that are involved in prion propagation.
  Four different anti-PrP antibodies that react with PrP^C on the cell surface inhibited PrP^ formation in cells persistently infected with prion. The antibody-PrP^C complex on the cell surface was not internalized efficiently and tended to retain on the cell surface. These results suggest that anti-PrP mAb antagonized PrP^ formation by interfering with the regular PrP^C degradation pathway. In addition, we screened synthesized sulfated glycosides for the inhibition of PrP^ formation and found 4-sulfo-N-acetylglucosamie and 6-sulfo-N-acetylglucosamine inhibited PrPSc formation in prion-infected cells. These sulfated glycosides accelerated the endocytosis of PrP^C and reduced a total amount of PrP^C, while sulfated glycosides that were not inhibited PrP^ formation did not reduce the total amount of PrP^C. These results indicated that sulfated glycosides and glycosaminoglycans inhibited PrP^ formation by facilitating the degradation of PrP^C.
  We established subclones of Neuro2a (N2a) mouse neuroblastoma cells and distinguished prion-susceptible and non-susceptible subclones. One non-susceptible subclone, N2a-1, expressed PrP^C as the same level as parental N2a and other prion-susceptible subclones. There was no difference in the binding of PrP^ to the susceptible and non-susceptible subclones. Presence of N2a-1, which expresses PrP^C but is resistance to prion propagation, indicated the involvement of host factors other than PrP^C in prion propagation. To identify such host factors, the gene expression profiles between N2a subclones were analyzed by DNA microarray. We selected 36 and 18 genes, which expressed more than two-fold in prion susceptible subclone N2a-5 and prion resistant subclone N2a-1, respectively. We assessed the influence of these genes on prion susceptibility by reducing the gene expression by siRNA technique and found that siRNA against F2, Al, and C5 genes inhibited prion propagation in prion-infected N2a-5 cells.
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (A), HOKKAIDO UNIVERSITY, Principal investigator, Competitive research funding, 15208029
• 免疫組織化学的手法によるプリオン蛋白の迅速検出法並びに検出感度に関する研究
  科学研究費助成事業 萌芽研究
  2002 - 2003
  古岡 秀文; 堀内 基広; 松井 高峯; 古林 与志安; 品川 森一
  (1)プリオン蛋白の免疫組織化学的検出感度に関する研究
  昨年度までに開発した免疫組織化学的高感度法とWB法の感度の比較を行った.スクレイピー帯広株脳内接種マウスを用いて経時的なプリオン蛋白の検出により比較を行った.WB法では30日目で脾臓に,60〜90日目に中枢神経系に異常プリオン蛋白が検出できた.一方,免疫組織化学的には脾臓は90日目,中枢神経系では120日目に検出可能であった.WB法ではマウス脾臓全量からの検出であるのに対して,免疫組織化学では約5μmの組織切片を使うことから単純な感度比較はできないが,免疫組織化学的方法は検出感度の点ではWB法に比較して劣ることが明らかになった.定量的な解析は現在実施中である.
  (2)パネル抗体による免疫組織化学コアエピトープに関する研究
  市販されている抗体を含め,これまでに開発された17種類の抗体を用いて,抗体の種特異性について,抗体エピトープと動物種間のアミノ酸配列との関連から検討した.動物種のアミノ酸配列と抗体エピトープの不一致が抗体の種特異性を規定する場合と規定しない場合があった.特異性を規定しない場合には,動物種間差でみられるいくつかのアミノ酸の違いは反応性を左右しない.しかしながら,アミノ酸一つの違いが動物種差としての特異性を規定する場合や,同じ領域を認識する抗体にも反応性に差がみられた.このことから,免疫組織化学的抗原抗体反応は抗体のエピトープ領域全てが関与するのではなく,限定されたコア領域に依存する可能性があることが推察された.
  日本学術振興会, 萌芽研究, 帯広畜産大学, Coinvestigator not use grants, Competitive research funding, 14656118
• 非通常病原体プリオンの分子機構
  科学研究費助成事業 基盤研究(C)
  2002 - 2002
  片峰 茂; 堂浦 克美; 金子 清俊; 小野寺 節; 福岡 伸一; 堀内 基広
  本邦におけるプリオン研究者の情報交換の促進と将来の共同研究プロジェクト立ち上げのための準備を目的に本研究を遂行した。情報交換に関しては、平成14年10月21日に長崎において班会議を開催し、班員に加えて数名の内外のプリオン研究者による情報交換と討議の場をもった。その結果、個々の班員間の往来及び研究材料の共有などのいくつかの実が挙がっている。例えば、片峰と小野寺は各々が開発したプリオン蛋白遺伝子に関わる遺伝子改変マウスと培養細胞株を共有することにより、プリオン病神経変性の機構解明へ向けた共同研究の進展が図られた。
  準備研究に関しては、プリオン研究進展に極めて大きな意味をもつ種々のモデル動物、細胞株、抗体、解析システムの開発が行われ、将来の大型共同研究プロジェクトへの準備は整ったと考えられる。以下に特筆される成果を挙げる。
  (1)プリオン持続感染細胞株の樹立(片峰)
  (2)プリオン類似蛋白(Dpl)遺伝子トランスジェニックマウスの樹立(片峰)
  (3)プリオン蛋白(PrP)と相互作用をする分子の同定法の開発(堂浦)
  (4)異常プリオン蛋白(PrPSc)に特異的立体構造を認識する抗体の確立(堀内)
  (5)不死化によるPrP欠損神経細胞株の樹立(小野寺)
  (6)PrPの細胞内挙動の顕微鏡下での追跡法の確立(金子)
  (7)タンパク質の2次構造変換定理の発見(柳川)
  (8)微量核酸(RNA)同定法の開発(福岡)
  本年度は、他領域との重複などの問題点があり、新規特定領域への申請は見送ったが、本研究の成果を基礎に来年度以降の申請へむけ、さらなる体制整備を行う予定である。
  日本学術振興会, 基盤研究(C), 長崎大学, Coinvestigator not use grants, Competitive research funding, 14607004
• プリオンの増殖を規定する正常型-異常型プリオン蛋白質の分子間相互作用に関する研究
  科学研究費助成事業 特定領域研究(C)
  2001 - 2002
  堀内 基広
  病原性プリオン蛋白質(PrPSc)と正常型プリオン蛋白質(PrPC)が直接会合することが、PrPScの増殖の第一段階である。そこで、PrPCとPrPScの結合に関与するPrP分子上のドメインを調べることを目的として、各種PrP合成ペプチドがPrPC-PrPSc間の相互作用を阻害するか否かについて検討した。PrPCがPrPSc存在下でPrPSc様のProteinase K(PK)抵抗性分子(PrP-res)に転換する試験管内転換反応で、PrP合成ペプチドaa117-141、aa166-179、aa200-223はPrPCがPrP-resに転換するのを阻害した。これらのPrPペプチドはPrPCと結合することにより、PrPCとPrPScの結合を阻害することが判明した。結合阻害活性を示すPrPペプチドは反応条件下ではβシート構造をとる傾向があることから、PrPペプチドがPrPScのPrPCへの結合ドメインを模倣することでPrPCと結合し、PrPC上のPrPSc結合ドメインを占拠する結果、PrPCとPrPScの結合を阻害する可能性が示唆された。
  PrPCとPrPScの分子間相互作用を解析する道具として、PrP分子に対するモノクローナル抗体(mAb)パネルを作製した。計34のmAbは認識するエピトープから9群に分類された。グループI〜VIはPrP分子上のリニアエピトープを認識する抗体、グル-プVII、VIIIはPrP分子上の構造エピトープを認識する抗体であった。また、グループIXはPrPSc特異的エピトープを認識する抗体である可能性が強く示唆された。aa59-89のリニアエピトープを認識するmAb、aa143-151を認識するmAb、およびaa1555-231領域内の構造エピトープを認識するmAbはプリオン感染細胞におけるPrPScの合成を阻害した。これらの抗体は細胞膜上に発現するPrPCと強く反応することから、抗体が細胞膜上のPrPCと結合することで正常なPrPC代謝経路が影響を受けるために、PrPSc合成の基質となるPrPCの供給が阻害された結果であると考えられる。
  日本学術振興会, 特定領域研究(C), 帯広畜産大学, Principal investigator, Competitive research funding, 13226004
• Studies on the entry ports for the scrapie agent (prion)
  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
  2000 - 2002
  HORIUCHI Motohiro; TANABE Shigeyuki; FORUOKA Hidefumi
  Major cause of infection in animal prion diseases is thought to be consumption of prion-contaminated stuff orally. There is evidence that the enteric nerve system (ENS) and lymphoid tissues, especially gut-associated lymphoid tissues (GATL) is involved in the infection of prion via alimentary tract. To elucidate the initial entry port for prion, we inoculated prion to alympholasia (aly) mice that show a deficiency in the systemic lymph nodes and Peyer's patches with various route. The aly/aly mice was susceptible to prion by intra-cranial inoculation, however, they showed reduced susceptibility with intra-peritoneal inoculation. The aly/aly mice were completely resistant to with per os administration, while C57BL/6J mice, the wild type mice for aly/aly mice, were sensitive to per os administration as they got the terminal stage of disease 307±7 days post inoculation. The prion infectivities were detected in the intestine and spleen of prion-inoculated C57BL/6J mice even after the early stage of exposure, whereas no infectivity was detected from those tissues of prion-inoculated aly/aly mice. PrPSc also detected in the intestine and spleen only of C57BL/6 mice, however PrPSc was not detected in the spleen and intestine of aly/aly mice that was affected by scrapie with the intra-peritoneal inoculation. No apparent difference in the organization of enteric nerve system was found between wild type and aly/aly mice. These results indicate that not ENS but GALT acts as a primary entry port for prion after the oral exposure.
  Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Obihiro University of Agriculture and Veterinary Medicine, Principal investigator, Competitive research funding, 12660268
• Etiological and molecular genetic surveillance of scrapie in Mongolia
  Grants-in-Aid for Scientific Research(基盤研究(B))
  2000 - 2002
  品川 森一; Motohiro HORIUCHI; 堀内 基広; 石黒 直隆; 村松 康和
  Mongolia has a far long history for sheep-farming and is now farming over twelve million sheep. In the future, Mongolia has a potential for the major provider of livestock products among the world. Thus surveillance of prion diseases in animal in Mongolia will provide important information to evaluate the safety of livestock products in the country. During the research term, we visit to central, west, and east part of Mongolia, we did not get a positive indication on the existence of scrapie-suspected sheep in hearing. We also checked PrPSc in the medulla oblongata from sheep showed neurolo...
  Ministry of Education, Culture, Sports, Science and Technology, 基盤研究(B), 帯広畜産大学, Principal investigator, Competitive research funding, 12575030
• Basic studies on the biosynthesis of pathogenic prion protein
  Grants-in-Aid for Scientific Research(基盤研究(B))
  2000 - 2002
  品川 森一; Sou-ichi MAKINO; 牧野 壮一; 堀内 基広; 古岡 秀文
  An abnormal isoform of prion protein (PrPSc) is a major component of the infectious agent "prion" and the biosynthesis of PrPSc play a central role in pathogenesis of prion diseases. Once prion enters the host, PrPSc binds to normal prion protein (PrPC) and converts PrPC into PrPSc. During the conformational transformation, PrPSc acts as a template or seed. In this study, we analyzed inter-molecular interaction between PrPC and PrPSc that is a key event in PrPSc generation.We found that PrPC bound to heterologous PrPSc but did not convert to PrPSc and that the conversion reaction could be i...
  Ministry of Education, Culture, Sports, Science and Technology, 基盤研究(B), 帯広畜産大学, Coinvestigator not use grants, Competitive research funding, 12460130
• Studies on diversity of sheep scrapie agents
  Grants-in-Aid for Scientific Research(基盤研究(C))
  1997 - 1999
  堀内 基広; Takane MATSUI; 牧野 壮一; 石黒 直隆
  The agent (prion) of transmissible spongiform encephalopathies (prion diseases) including scrape in sheep, bovine spongiform encephalopathy, and Creutzfeldt-Jakob disease (CJD) in human does not possess the agent-specific nucleic acid as a genome. Therefore genome typing and/or antigenic typing are not applicable for distinction of prion strains. However, prion strains can be distinguishable to some extent by biological and biochemical properties. In this study, we attempted to elucidate whether diversity of sheep prion strains exist in Japan. In addition, we analyzed genetic factor(s) othe...
  Ministry of Education, Culture, Sports, Science and Technology, 基盤研究(C), 帯広畜産大学, Principal investigator, Competitive research funding, 09660312
• In Vitro studies on the formation of prion amyloid
  Grants-in-Aid for Scientific Research(基盤研究(B))
  1996 - 1998
  Morikazu SHINAGAWA; 堀内 基広; 桑山 秀人; 石黒 直隆
  As a final goal of this study is in vitro formation of infectious prion amyloid using mouse PrP^C, in vitro structural conversion of PrP^C using a small amount of mouse prion was carried out. To eliminate the effects of unknown mouse protein contaminating in PrP^C, mouse recombinant PrP^C which possessed a histidine tag at N-terminus of mature PrP^C was used. The recombinant PrP^C converted to a proteinase K (PK) resistant form in the presence of one-thousandth amounts of mouse scrapie prion at pH 5.2 and room temperature for one day and after 14 days the PK-resistance increased more. A dec...
  Ministry of Education, Culture, Sports, Science and Technology, 基盤研究(B), 帯広畜産大学, Coinvestigator not use grants, Competitive research funding, 08456145
• Development of sensitive diagnostic methods for transmissible spongiform encephalopathies.
  Grants-in-Aid for Scientific Research(試験研究(B), 基盤研究(A))
  1995 - 1997
  Morikazu SHINAGAWA; 中川 迪夫; 堀内 基広; 久保 正法; 山田 明夫; 古岡 秀文
  In order to increase the sensitivity for detection of prion protein, the method to prepare samples from tissues for Western blotting was reformed. In mouse scrapie model, prion protein could be detected in the samples prepared from the spleen 1 week after intraperitoneal infection by the new method. It is to be desirable that more convenient method than Western blotting to examine many samples for the presence of prion protein. We developed an ELISA whose sensitivity was about two times higher than that of Western blotting using mouse model. We first confirmed that this ELISA method could b...
  Ministry of Education, Culture, Sports, Science and Technology, 試験研究(B), 基盤研究(A), 帯広畜産大学, Coinvestigator not use grants, Competitive research funding, 07556068
• 伝達性海綿状脳症関連蛋白質PrPによる神経細胞死の分子機構に関する研究
  科学研究費補助金(奨励研究(A))
  1995 - 1995
  堀内 基広
  本研究は伝達性海綿状脳症における神経細胞死と本病関連蛋白質(PrP)の関係を分子レベルで解明することを目的として開始した。これまでにマウス神経芽腫細胞(N2a)、およびラット副腎クロム親和性細胞腫(PC12)に羊PrPC(PrPCは正常型PrPを示す)を過発現させることに成功しているので、これら培養細胞を用いて研究を行った。羊PrPC過発現細胞は内在性PrPCに比べ数十倍羊PrPCを発現している。羊PrPCの過発現自体が細胞に及ぼす影響を調べたが、NGF、cAMPの刺激に対する応答などの生物性状にコントロールとなる正常細胞との間に差は認められなかった。また、羊PrPCの過発現自体が細胞の活性に影響するか否かをLDH遊離試験、Ho33258核染色により調べたが、正常細胞との間に差は認められなかった。クロロキン処理により羊PrPCの蓄積を高めた場合でも顕著な差は認められなかった。神経細胞毒性が報告されているマウスPrPコドン106-129の合成ペプチドに相応する羊PrP合成ペプチドを羊PrPC過発現細胞に添加し本ペプチドの細胞致死活性を調べたが、羊PrP合成ペプチドによる羊PrPC過発現細胞の選択的な細胞死は観察されなかった。現在まで、PrPによる神経細胞毒性が観察されたのはマウスおよびラットの初代神経培養細胞に限られており、株化細胞での例はない。本研究においても神経系株化細胞で...
  文部科学省, 奨励研究(A), 帯広畜産大学, Principal investigator, Competitive research funding, 07760272
• Study on the role of a host protein, PrP,in scrapie.
  Grants-in-Aid for Scientific Research(一般研究(B))
  1992 - 1994
  Morikazu SHINAGAWA; 堀内 基広
  1) Infectious amyloid, which is thought to be an etiological agent of scrapie, consists of PrP^, an isoform of a host membrane protein, PrP^C, which is highly expressed in the central nervous system (CNS). It supposedly is one cause of high accumulation of the infectious amyloid in the CNS.We have established a L-929 derived cell-line expressing exogenous PrP (L-BPrP3) by introducing bovine PrP cDNA.In this study, scrapie replication was examined in this cell-line and in the L-929 cell-line. We expected that L-BPrP3 supports sheep scrapie, however, the replication of sheep scrapie in th...
  Ministry of Education, Culture, Sports, Science and Technology, 一般研究(B), 帯広畜産大学, Coinvestigator not use grants, Competitive research funding, 04454113
• 猫パルボウイルス亜群の分子進化と犬パルボウイルス出現機構に関する研究
  科学研究費補助金(奨励研究(A))
  1993 - 1993
  堀内 基広
  本研究は、猫パルボウイルス亜群に属する猫汎白血球減少症ウイルス(FPLV)、ミンク腸炎ウイルス(MEV)、犬パルボウイルス(CPV)の分子遺伝学的な解析を行い、これら3ウイルスの相互関係、ならびにCPV出現機構の解明を目的とした。ウイルスのVPgeneに存在する宿主域決定領域(60-91map units, Horiuchi et al.,J.Gen.Virol.,in press)をPCR法により増幅し、直接塩基配列決定法(Higuchi and Ochman,Nucleic Acide Res.,1989)により塩基配列を決定した。現在までに、FPLV2株(日本;1974,フランス;分離年不明)、CPV4株(日本;1979,1982,1984,1991)、MEV1株(日本,1980)の計7株の塩基配列を決定した。このうち代表的なもの(FPLV1株,CPV3株,MEV1株)についてはDNA Data Base of Japan(DDBJ)に登録した。株数が少なかったため分子系統樹を書くには至らなかったが、塩基配列より予想されるアミノ酸配列の多重アライメントの結果、宿主域決定領域内に系統発生的にCPV特異的と考えられる5個のアミノ酸を同定した[amino acid(aa)93,aa103,aa305,aa564,aa568]。また、Parrishらが米国で報告したaa300,...
  文部科学省, 奨励研究(A), 帯広畜産大学, Principal investigator, Competitive research funding, 05760217
• Survey on scrapie in Japanese : detection of PrP^ and studies on the distribution of the PrP genotypes in sheep.
  Grants-in-Aid for Scientific Research(総合研究(A))
  1991 - 1993
  Morikazu SHINAGAWA; 石黒 直隆; 平井 克也; 本多 英一; 見上 彪; 堀内 基広; 小沼 操
  1. Restriction fragment length polymorphism (RFLP) on the sheep-PrP gene : To investigate the RFLP types on the PrP-gene, chromosomal DNAs from sheep tissues collected from various districts in Japan were analyzed using restriction endonucleases EcoRI and HindIII and the coding region of sheep PrP as a probe. Sheep were classified into 6 types. Types I to III corresponded to the types reported in England. Nearly half of the scrapie sheep belonged to type I and the frequency of type II in scrapie sheep was low. Distribution of the types in different parts of Japan showed some variations.2. S...
  Ministry of Education, Culture, Sports, Science and Technology, 総合研究(A), 帯広畜産大学, Coinvestigator not use grants, Competitive research funding, 03304025
• Role of Host-coded Protein "PrP"in Scrapie : Preparation of Transformed Cells with Heterologous or Artificially Mutated PrP Gene.
  Grants-in-Aid for Scientific Research(一般研究(B))
  1990 - 1991
  Morikazu SHINAGAWA; 堀内 基広; 石黒 直隆
  A cDNA clone encoding bovine scrapie-associated fibril protein, PrP, from a bovine brain cDNA library and 6 amplified genomic DNA clones of bovine PrP were characterized. These clones possessed specific characteristics observed in other animal PrP genes. However, the bovine PrP was divided into two types by the number of repeats. One possessed four octapeptide repetitive sequences like other animal PrP genes and consisted of 256 amino acids, the other had five such repetitive sequences and 264 amino acids. The amino acid sequence of the former bovine PrP agreed with that of sheep PrP up to ...
  Ministry of Education, Culture, Sports, Science and Technology, 一般研究(B), 帯広畜産大学, Coinvestigator not use grants, Competitive research funding, 02454097
• Isolation and molecular characterization of bovine T-cell receptor genes
  Grants-in-Aid for Scientific Research(一般研究(C))
  1990 - 1991
  Naotaka ISHIGURO; 堀内 基広
  The nucleotide sequences of bovine T-cell receptor (TCR ; alpha, beta, gamma, and delta chains) genes which from cDNA libraries constructed from bovine peripheral blood lymphocytes and thymic lymphocytes were determined.1. Of 30 positive cDNA clones for bovine TCR alpha chain, 20 contained complete and correct rearrangements, whereas 4 contained only one Constant (C) region, and 6 were nonfunctional because of incomplete rearrangement. The bovine Calpha gene is composed of 140 amino acid residues with striking similarity to the Ca region of human and mouse.2. Of 38 CDNA clones for TCR beta ...
  Ministry of Education, Culture, Sports, Science and Technology, 一般研究(C), 帯広畜産大学, Coinvestigator not use grants, Competitive research funding, 02806055
• 動物パルボウイルス遺伝子のウイルス種を規定する遺伝子領域に関する研究
  科学研究費助成事業
  1990 - 1990
  堀内 基広
  日本学術振興会, 奨励研究(A), 帯広畜産大学, 02856082
• Development of Diagnostic Method for Unconventional Agent (Scrapie Gagent) Infection.
  Grants-in-Aid for Scientific Research(試験研究, 試験研究(B))
  1988 - 1990
  Morikazu SHINAGAWA; 吉野 智男; 太田 千佳子; 石黒 直隆; 百渓 英一; 堀内 基広
  The usefulness of detecting the protein (PrP) consisting of scrapie-associated fibrils in the lymphoreticular organs of sheep as a diagnostic tool was investigated. The PrP was detected by means of a rabbit-anti-sheep PrP polyclonal antibody by Western blot analysis. PrP was detected in samples from the central nervous system of five of six sheep showing clinical signs of natural scrapie infection, in spleen samples from four of the six sheep and in lymph node samples taken from three of the sheep. PrP was detected in the spleen and lymph node samples, but not in the central nervous system ...
  Ministry of Education, Culture, Sports, Science and Technology, 試験研究, 試験研究(B), 帯広畜産大学, Coinvestigator not use grants, Competitive research funding, 63860040