基本情報

所属

• 理学研究院　生物科学部門　生殖発生生物学分野

職名

• 教授

学位

• 博士（農学）（名古屋大学）

ホームページURL

https://sites.google.com/site/kuroiwagroup/
https://sites.google.com/site/kuroiwagroup/home/english-page

J-Global ID

• 200901001109628284

プロフィール

• 哺乳類、鳥類を対象に、性決定の分子メカニズムの解明を目指しています。Y染色体をもたず、哺乳類の性決定遺伝子であるSRY遺伝子も失っているアマミトゲネズミにおいて、SRY遺伝子なしにどのように性が決定されるのか、どのようにY染色体の消失が起きたのかを明らかにしようとしています。また、性染色体がZZでオス、ZWでメスとなる鳥類では、オスの精巣分化がどのような分子メカニズムで起きるのか、特に哺乳類との違いに着目して研究を進めています。また、ニワトリ、ニホンウズラ、エミューなどを用いて、W染色体上のメス決定遺伝子を見つけるために研究を進めています。

研究キーワード

• トゲネズミ   Y染色体   哺乳類   鳥類   性分化   性決定   性   性染色体   

研究分野

• ライフサイエンス / 発生生物学
• ライフサイエンス / 遺伝学

担当教育組織

•  学士課程　理学部
•  修士課程　生命科学院
•  博士課程　生命科学院

職歴

• 2016年04月 - 現在 北海道大学 大学院理学研究院生物科学部門 教授
• 2009年04月 - 2016年03月 北海道大学 大学院理学研究院生物科学部門 准教授
• 2008年11月 - 2009年03月 北海道大学 大学院理学研究院生物科学部門 講師
• 2005年04月 - 2008年10月 北海道大学 創成科学共同研究機構 講師
• 2003年10月 - 2005年03月 北海道大学 先端科学技術共同研究センター 講師
• 2003年04月 - 2003年09月 日本学術振興会 特別研究員PD
• 2001年04月 - 2003年03月 日本学術振興会 特別研究員DC2
• 2001年 - Researcher of Japan Society for the Promotion Science

学歴

• 1999年04月 - 2002年03月   名古屋大学   大学院生命農学研究科   博士後期課程 応用分子生命科学専攻
•         - 2002年   名古屋大学
• 1997年04月 - 1999年03月   名古屋大学   大学院生命農学研究科   博士前期課程 畜産学専攻
•         - 1999年   名古屋大学
• 1993年04月 - 1997年03月   名古屋大学   農学部   資源生物環境学科
•         - 1997年   名古屋大学

所属学協会

• 日本染色体学会   日本実験動物学会   日本遺伝学会   日本分子生物学会   

研究活動情報

論文

• Diethylstilbestrol reduces primordial germ cells in male Japanese quail

  Mizushima S, Kuroiwa A

  Poultry Science 102 10 102910 - 102910 2023年10月 [査読有り][通常論文]
  
• The neo-X does not form a Barr body but shows a slightly condensed structure in the Okinawa spiny rat (Tokudaia muenninki).

  Ryoma Kudo, Ikuya Yoshida, Luisa Matiz Ceron, Shusei Mizushima, Yoko Kuroki, Takamichi Jogahara, Asato Kuroiwa

  Cytogenetic and genome research 2023年06月02日 

  X chromosome inactivation (XCI) is an essential mechanism for gene dosage compensation between male and female cells in mammals. The Okinawa spiny rat (Tokudaia muenninki) is a native rodent in Japan with XX/XY sex chromosomes, like most mammals; however, the X chromosome has acquired a neo-X region (Xp) by fusion with an autosome. We previously reported that dosage compensation has not yet evolved in the neo-X region; however, X-inactive-specific transcript (Xist) RNA (long non-coding RNA required for the initiation of XCI) is partially localized in the region. Here, we show that the neo-X region represents an early chromosomal state in the acquisition of XCI by analyses of heterochromatin and Barr body formation. We found no evidence for heterochromatin formation in the neo-X region by RBA (R-banding by acridine orange) assays and immunostaining of H3K27me3. Double-immunostaining of H3K27me3 and HP1, a component of the Barr body, revealed that the entire ancestral-X chromosome region (Xq) showed a bipartite folded structure. By contrast, HP1 was not localized to the neo-X region. However, BAC FISH revealed that the signals of genes on the neo-X region of the inactive X chromosome were concentrated in a narrow region. These findings indicated that although the neo-X region of the inactive X chromosome does not form a complete Barr body structure (e.g., it lacks HP1), it forms a slightly condensed structure. These findings combined with the previously reported partial binding of Xist RNA suggest that the neo-X region exhibits incomplete inactivation. This may represent an early chromosomal state in the acquisition of the XCI mechanism.
• Current Approaches to and the Application of Intracytoplasmic Sperm Injection (ICSI) for Avian Genome Editing

  Shusei Mizushima, Tomohiro Sasanami, Tamao Ono, Asato Kuroiwa

  Genes 14 3 757 - 757 2023年03月20日 

  Poultry are one of the most valuable resources for human society. They are also recognized as a powerful experimental animal for basic research on embryogenesis. Demands for the supply of low-allergen eggs and bioreactors have increased with the development of programmable genome editing technology. The CRISPR/Cas9 system has recently been used to produce transgenic animals and various animals in the agricultural industry and has also been successfully adopted for the modification of chicken and quail genomes. In this review, we describe the successful establishment of genome-edited lines combined with germline chimera production systems mediated by primordial germ cells and by viral infection in poultry. The avian intracytoplasmic sperm injection (ICSI) system that we previously established and recent advances in ICSI for genome editing are also summarized.
• Turnover of mammal sex chromosomes in the Sry-deficient Amami spiny rat is due to male-specific upregulation of Sox9

  Miho Terao, Yuya Ogawa, Shuji Takada, Rei Kajitani, Miki Okuno, Yuta Mochimaru, Kentaro Matsuoka, Takehiko Itoh, Atsushi Toyoda, Tomohiro Kono, Takamichi Jogahara, Shusei Mizushima, Asato Kuroiwa

  Proceedings of the National Academy of Sciences 119 49 e2211574119  2022年11月28日 [査読有り][通常論文]
   

  Mammalian sex chromosomes are highly conserved, and sex is determined by SRY on the Y chromosome. Two exceptional rodent groups in which some species lack a Y chromosome and Sry offer insights into how novel sex genes can arise and replace Sry, leading to sex chromosome turnover. However, intensive study over three decades has failed to reveal the identity of novel sex genes in either of these lineages. We here report our discovery of a male-specific duplication of an enhancer of Sox9 in the Amami spiny rat Tokudaia osimensis, in which males and females have only a single X chromosome (XO/XO) and the Y chromosome and Sry are completely lost. We performed a comprehensive survey to detect sex-specific genomic regions in the spiny rat. Sex-related genomic differences were limited to a male-specific duplication of a 17-kb unit located 430 kb upstream of Sox9 on an autosome. Hi-C analysis using male spiny rat cells showed the duplicated region has potential chromatin interaction with Sox9. The duplicated unit harbored a 1,262-bp element homologous to mouse enhancer 14 (Enh14), a candidate Sox9 enhancer that is functionally redundant in mice. Transgenic reporter mice showed that the spiny rat Enh14 can function as an embryonic testis enhancer in mice. Embryonic gonads of XX mice in which Enh14 was replaced by the duplicated spiny rat Enh14 showed increased Sox9 expression and decreased Foxl2 expression. We propose that male-specific duplication of this Sox9 enhancer substituted for Sry function, defining a novel Y chromosome in the spiny rat.
• Analysis of Sex Chromosome Evolution in the Clade Palaeognathae from Phased Genome Assembly.

  Okuno M, Mizushima S, Kuroiwa A, Itoh T

  Genome Biol Evol 13 11 evab242  2021年11月 [査読有り][通常論文]
  
• Inositol-1,4,5-Trisphosphate Receptor-1 and -3 and Ryanodine Receptor-3 May Increase Ooplasmic Ca2+ During Quail Egg Activation

  Shusei Mizushima, Tomohiro Sasanami, Tamao Ono, Norio Kansaku, Asato Kuroiwa

  The Journal of Poultry Science 59 2 175 - 181 2021年08月 [査読有り][通常論文]
  
• Cyclin D1 gene expression is essential for cell cycle progression from the maternal-to-zygotic transition during blastoderm development in Japanese quail.

  Shusei Mizushima, Tomohiro Sasanami, Tamao Ono, Mei Matsuzaki, Norio Kansaku, Asato Kuroiwa

  Developmental biology 476 249 - 258 2021年08月 [招待有り]
   

  Embryogenesis proceeds by a highly regulated series of events. In animals, maternal factors that accumulate in the egg cytoplasm control cell cycle progression at the initial stage of cleavage. However, cell cycle regulation is switched to a system governed by the activated nuclear genome at a specific stage of development, referred to as maternal-to-zygotic transition (MZT). Detailed molecular analyses have been performed on maternal factors and activated zygotic genes in MZT in mammals, fishes and chicken; however, the underlying mechanisms remain unclear in quail. In the present study, we demonstrated that MZT occurred at blastoderm stage V in the Japanese quail using novel gene targeting technology in which the CRISPR/Cas9 and intracytoplasmic sperm injection (ICSI) systems were combined. At blastoderm stage V, we found that maternal retinoblastoma 1 (RB1) protein expression was down-regulated, whereas the gene expression of cyclin D1 (CCND1) was initiated. When a microinjection of sgRNA containing CCND1-targeted sequencing and Cas9 mRNA was administered at the pronuclear stage, blastoderm development stopped at stage V and the down-regulation of RB1 did not occur. This result indicates the most notable difference from mammals in which CCND-knockout embryos are capable of developing beyond MZT. We also showed that CCND1 induced the phosphorylation of the serine/threonine residues of the RB1 protein, which resulted in the degradation of this protein. These results suggest that CCND1 is one of the key factors for RB1 protein degradation at MZT, and the elimination of RB1 may contribute to cell cycle progression after MZT during blastoderm development in the Japanese quail. Our novel technology, which combined the CRISPR/Cas9 system and ICSI, has the potential to become a powerful tool for avian-targeted mutagenesis.
• Diversification of mineralocorticoid receptor genes in a subterranean rodent, the naked mole-rat.

  Kaori Oka, Hidemasa Bono, Asato Kuroiwa, Shusuke Fujioka, Atsushi Shimizu, Yoshinao Katsu, Kyoko Miura

  Journal of molecular endocrinology 66 4 299 - 311 2021年05月11日 [招待有り]
   

  Naked mole-rats (Heterocephalus glaber) inhabit subterranean burrows in savannas and are, thus, unable to access free water. To identify their mechanism of osmoregulation in xeric environments, we molecularly cloned and analyzed the nuclear receptor subfamily 3 group C member 2 (NR3C2) gene encoding the mineralocorticoid receptor (MR), required for hormone-dependent regulation of genes contributing to body fluid homeostasis. Most vertebrates harbor a single MR homolog. In contrast, we discovered that MR is duplicated in naked mole-rats. The amino acid sequence of naked mole-rat MR1 is 90% identical to its mouse ortholog, and MR1 is abundantly expressed in the kidney and the nervous system. MR2 encodes a truncated protein lacking DNA- and ligand-binding domains of MR1 and is expressed in diverse tissues. Although MR2 did not directly transactivate gene expression, it increased corticosteroid-dependent transcriptional activity of MR1. Our results suggest that MR2 might function as a novel regulator of MR1 activity to fine-tune MR signaling in naked mole-rats.
• Expression profiling of sexually dimorphic genes in the Japanese quail, Coturnix japonica.

  Miki Okuno, Shuntaro Miyamoto, Takehiko Itoh, Masahide Seki, Yutaka Suzuki, Shusei Mizushima, Asato Kuroiwa

  Scientific Reports 10 1 20073 - 20073 2020年11月30日 [査読有り][通常論文]
   

  Research on avian sex determination has focused on the chicken. In this study, we established the utility of another widely used animal model, the Japanese quail (Coturnix japonica), for clarifying the molecular mechanisms underlying gonadal sex differentiation. In particular, we performed comprehensive gene expression profiling of embryonic gonads at three stages (HH27, HH31 and HH38) by mRNA-seq. We classified the expression patterns of 4,815 genes into nine clusters according to the extent of change between stages. Cluster 2 (characterized by an initial increase and steady levels thereafter), including 495 and 310 genes expressed in males and females, respectively, contained five key genes involved in gonadal sex differentiation. A GO analysis showed that genes in this cluster are related to developmental processes including reproductive structure development and developmental processes involved in reproduction were significant, suggesting that expression profiling is an effective approach to identify novel candidate genes. Based on RNA-seq data and in situ hybridization, the expression patterns and localization of most key genes for gonadal sex differentiation corresponded well to those of the chicken. Our results support the effectiveness of the Japanese quail as a model for studies gonadal sex differentiation in birds.
• トクノシマトゲネズミ（Tokudaia tokunoshimensis）の生息記録と2005年～2016年の分布

  城ヶ原貴通, 中家雅隆, 池村茂, 越本知大, 坂本信介, 橋本琢磨, 三谷匡, 黒岩麻里, 山田文雄

  哺乳類科学 60 1 105 - 116 日本哺乳類学会 2020年01月 [査読有り][通常論文]
   

  トゲネズミ属（Tokudaia）は琉球列島にのみ分布する固有属であり，トクノシマトゲネズミ（Tokudaia tokunoshimensis）は徳之島にのみ分布する固有種で，絶滅が危惧されている．本種の分布・生息状況についてはまとまった記録が蓄積されておらず，保全策を検討する上で分布・生息状況を明らかにする必要がある．本研究ではトクノシマトゲネズミの保全に向けた第一段階として本種の分布状況を明らかにするため，2011年から2015年の捕獲調査，自動撮影カメラによる調査に加え，2005年から2016年の目撃記録・直接観察の情報の整理ならびにその他の分布情報を精査した．その結果，トクノシマトゲネズミの分布は島北部の天城岳ならびに島中東部に位置する井之川岳周辺を分布の中心地域とし，南部の犬田布岳周辺にわずかに生息していることが明らかとなった．また，比較的良好な森林があるにも関わらず生息が確認できない地域が存在することが明らかとなった．本調査結果は，絶滅のおそれのあるトクノシマトゲネズミの保全地域の設定およびイエネコ（Felis catus）による捕食対策など生息地保全を含むさまざまな保全策の策定に基礎情報を提供するものである．

• Spiny rat SRY lacks a long Q-rich domain and is not stable in transgenic mice.

  Ogata Y, Nishikata M, Kitada K, Mizushima S, Jogahara T, Kuroiwa A

  Developmental Dynamics 248 9 784 - 794 2019年09月 [査読有り][通常論文]
  
• Regulation of the Sox3 Gene in an X0/X0 Mammal without Sry, the Amami Spiny Rat, Tokudaia osimensis.

  Kohei Washio, Shusei Mizushima, Takamichi Jogahara, Asato Kuroiwa

  Cytogenetic and genome research 159 3 143 - 150 2019年 [査読有り][通常論文]
   

  Two species of spiny rats, Tokudaia osimensis and Tokudaia tokunoshimensis, show an X0/X0 sex chromosome constitution due to the lack of a Y chromosome. The Sry gene has been completely lost from the genome of these species. We hypothesized that Sox3, which is thought to be originally a homologue of Sry, could function in sex determination in these animals in the absence of Sry. Sox3 was localized in a region of the X chromosome in T. osimensis homologous to mouse. A similar testis- and ovary-specific pattern of expression was observed in mouse and T. osimensis. Although the sequence of the Sox3 gene and its promoter are highly conserved, a 13-bp deletion was specifically found in the promoter region of the 2 spiny rat species. Reporter gene assays were performed to examine the effect of the 13-bp deletion in the promoter region on Sox3 regulation. Although an approximately 60% decrease in activity was observed using the Tokudaia promoters with the 13-bp deletion, the activity was recovered using a mutated promoter in which the deletion was filled with mouse sequence. To evaluate whether SOX3 could regulate Sox9 expression, a reporter gene assay was carried out using testis-specific enhancer of Sox9 core (TESCO). Co-transfection with a combination of mouse SF1 and mouse SOX3 or T. osimensis SOX3 resulted in a greater than 2-fold increase in activity of mouse and T. osimensis TESCO. These results support the idea that the function of SOX3 as a transcription factor, as has been reported in mice and humans, is conserved in T. osimensis. Therefore, we conclude that the Sox3 gene has no function in sex determination in Sry-lacking Tokudaia species.
• Sexual dimorphism in brain transcriptomes of Amami spiny rat (Tokudaia osimensis): a rodent species where males lack the Y chromosome

  Ortega MT, Johnson SA, Bivens N, Jogahara T, Kuroiwa A, Givan SA, Rosenfeld CS

  BMC Genomics 20 1 87  2019年 [査読有り][通常論文]
  
• Knockdown of DDX4 decreases the number of germ cells in male and female chicken embryonic gonads

  Aduma N, Izumi H, Mizushima S, Kuroiwa A

  Reproduction, Fertility and Development 2018年12月 [査読有り][通常論文]
  
• Yをもたない不思議な哺乳類―トゲネズミ

  黒岩 麻里

  実験医学 36 11 1938 - 1941 2018年07月 [査読有り][招待有り]
  
• Unique XCI evolution in Tokudaia: initial XCI of the neo-X chromosome in Tokudaia muenninki and function loss of XIST in Tokudaia osimensis

  Hideki Zushi, Chie Murata, Shusei Mizushima, Chizuko Nishida, Asato Kuroiwa

  CHROMOSOMA 126 6 741 - 751 2017年12月 [査読有り][通常論文]
   

  X chromosome inactivation (XCI) is an essential mechanism to compensate gene dosage in mammals. Here, we show that XCI has evolved differently in two species of the genus Tokudaia. The Amami spiny rat, Tokudaia osimensis, has a single X chromosome in males and females (XO/XO). By contrast, the Okinawa spiny rat, Tokudaia muenninki, has XX/XY sex chromosomes like most mammals, although the X chromosome has acquired a neo-X region by fusion with an autosome. BAC clones containing the XIST gene, which produces the long non-coding RNA XIST required for XCI, were obtained by screening of T. osimensis and T. muenninki BAC libraries. Each clone was mapped to the homologous region of the X inactivation center in the X chromosome of the two species by BAC-FISH. XIST RNAs were expressed in T. muenninki females, whereas no expression was observed in T. osimensis. The sequence of the XIST RNA was compared with that of mouse, showing that the XIST gene is highly conserved in T. muenninki. XIST RNAs were localized to the ancestral X region (Xq), to the heterochromatic region (pericentromeric region), and partially to the neo-X region (Xp). The hybridization pattern correlated with LINE-1 accumulation in Xq but not in Xp. Dosage of genes located on the neo-X chromosome was not compensated, suggesting that the neo-X region is in an early state of XCI. By contrast, many mutations were observed in the XIST gene of T. osimensis, indicating its loss of function in the XO/XO species.
• Flexible adaptation of male germ cells from female iPSCs of endangered Tokudaia osimensis

  Arata Honda, Narantsog Choijookhuu, Haruna Izu, Yoshihiro Kawano, Mizuho Inokuchi, Kimiko Honsho, Ah-Reum Lee, Hiroki Nabekura, Hiroshi Ohta, Tomoyuki Tsukiyama, Yasuhide Ohinata, Asato Kuroiwa, Yoshitaka Hishikawa, Mitinori Saitou, Takamichi Jogahara, Chihiro Koshimoto

  SCIENCE ADVANCES 3 5 e1602179  2017年05月 [査読有り][通常論文]
   

  In mammals, the Y chromosome strictly influences the maintenance of male germ cells. Almost all mammalian species require genetic contributors to generate testes. An endangered species, Tokudaia osimensis, has a unique sex chromosome composition XO/XO, and genetic differences between males and females have not been confirmed. Although a distinctive sex-determining mechanism may exist in T. osimensis, it has been difficult to examine thoroughly in this rare animal species. To elucidate the discriminative sex-determining mechanism in T. osimensis and to find a strategy to prevent its possible extinction, we have established induced pluripotent stem cells (iPSCs) and derived interspecific chimeras using mice as the hosts and recipients. Generated iPSCs are considered to be in the so-called "true naive" state, and T. osimensis iPSCs may contribute as interspecific chimeras to several different tissues and cells in live animals. Surprisingly, female T. osimensis iPSCs not only contributed to the female germ line in the interspecific mouse ovary but also differentiated into spermatocytes and spermatids that survived in the adult interspecific mouse testes. Thus, T. osimensis cells have high sexual plasticity through which female somatic cells can be converted to male germline cells. These findings suggest flexibility in T. osimensis cells, which can adapt their germ cell sex to the gonadal niche. The probable reduction of the extinction risk of an endangered species through the use of iPSCs is indicated by this study.
• Androgens and androgen receptor signaling contribute to ovarian development in the chicken embryo

  Ryoma Tanaka, Hiroe Izumi, Asato Kuroiwa

  MOLECULAR AND CELLULAR ENDOCRINOLOGY 443 C 114 - 120 2017年03月 [査読有り][通常論文]
   

  Androgens and androgen receptor (AR) signaling play important roles throughout development. In the chicken, AR signaling is involved in reproduction; however, its specific role is unclear. We show that AR signaling is involved in the normal development of the female embryonic gonads. The AR mRNA level was detected in male and female embryonic gonads by quantitative RT-PCR, and its expression was higher in females than in males at all developmental stages examined. In female embryos, the AR localized to nuclei of cells in the left gonad. Although AR expression was low in the majority of the medulla, high expression was detected in cells of lacunae within the medulla. In addition, AR expression increased in cells of cortical cords within the cortex with the progression of development. AR expression in the right gonad was lower than that in left gonad throughout development. In the male gonad, the AR localized to the cytoplasm of cells in seminiferous tubules at all stages. Female AR knockdown (ARKD) embryos infected with a retrovirus expressing micro RNAs targeting the AR showed normal asymmetric gonads (development of the left and depression of the right gonads), whereas the number of lacunae decreased. Furthermore, there was a disruption in the structure of cortical cords. By contrast, the gonads of ARKD males developed normally during embryogenesis. These results indicate that androgens and AR signaling are essential for the development of lacunae and cortical cords in gonads of female embryos. (C) 2017 Elsevier B.V. All rights reserved.
• Sex-determining mechanism in avians

  Asato Kuroiwa

  Advances in Experimental Medicine and Biology 1001 19 - 31 2017年 [査読有り][招待有り]
   

  The sex of birds is determined by inheritance of sex chromosomes at fertilization. The embryo with two Z chromosomes (ZZ) develops into a male by contrast, the embryo with Z and W chromosomes (ZW) becomes female. Two theories are hypothesized for the mechanisms of avian sex determination that explain how genes carried on sex chromosomes control gonadal differentiation and development during embryogenesis. One proposes that the dosage of genes on the Z chromosome determines the sexual differentiation of undifferentiated gonads, and the other proposes that W-linked genes dominantly determine ovary differentiation or inhibit testis differentiation. Z-linked DMRT1, which is a strong candidate avian sex-determining gene, supports the former hypothesis. Although no candidate W-linked gene has been identified, extensive evidence for spontaneous sex reversal in birds and aneuploid chimeric chickens with an abnormal sex chromosome constitution strongly supports the latter hypothesis. After the sex of gonad is determined by a gene(s) located on the sex chromosomes, gonadal differentiation is subsequently progressed by several genes. Developed gonads secrete sex hormones to masculinize or feminize the whole body of the embryo. In this section, the sex-determining mechanism as well as the genes and sex hormones mainly involved in gonadal differentiation and development of chicken are introduced.
• The cryptic Y-autosome translocation in the small Indian mongoose, Herpestes auropunctatus, revealed by molecular cytogenetic approaches

  Chie Murata, Hirohito Sawaya, Katsushi Nakata, Fumio Yamada, Issei Imoto, Asato Kuroiwa

  CHROMOSOMA 125 4 807 - 815 2016年09月 [査読有り][通常論文]
   

  In initial studies of the eutherian small Indian mongoose (Herpestes auropunctatus), the Y chromosome could not be identified in somatic cells. The male chromosome number is uniquely odd, 2n = 35, whereas that of females is 2n = 36. Previous reports indicated that this unique karyotype resulted from a translocation of the ancestral Y chromosome to an autosome. However, it has been difficult to identify the chromosomes that harbor the translocated Y chromosomal segment because it is an extremely small euchromatic region. Using a Southern blot analysis, we detected four conserved Y-linked genes, SRY, EIF2S3Y, KDM5D, and ZFY, in the male genome. We cloned homologues of these genes and determined their sequences, which showed high homology to genes in two carnivore species, cat and dog. To unambiguously identify the Y-bearing autosome, we performed immunostaining of pachytene spermatocytes using antibodies against SYCP3, gamma H2AX, and the centromere. We observed trivalent chromosomes, and the associations between the distal ends of the chromosomes were consistent with those of Y and X1 chromosomes. The centromere of the Y chromosome was located on the ancestral Y chromosomal segment. We mapped the complementary DNA (cDNA) clones of these genes to the male chromosomes using fluorescence in situ hybridization (FISH), and the linear localization of all genes was confirmed by two-colored FISH. These Y-linked genes were localized to the proximal region of the long arm of a single telomeric chromosome, and we successfully identified the chromosome harboring the ancestral Y chromosomal segment.
• Ancestral Y-linked genes were maintained by translocation to the X and Y chromosomes fused to an autosomal pair in the Okinawa spiny rat Tokudaia muenninki

  Chie Murata, Yoko Kuroki, Issei Imoto, Asato Kuroiwa

  CHROMOSOME RESEARCH 24 3 407 - 419 2016年09月 [査読有り][通常論文]
   

  Two species of the genus Tokudaia lack the Y chromosome and SRY, but several Y-linked genes have been rescued by translocation or transposition to other chromosomes. Tokudaia muenninki is the only species in the genus that maintains the Y owing to sex chromosome-autosome fusions. According to previous studies, many SRY pseudocopies and other Y-linked genes have evolved by excess duplication in this species. Using RNA-seq and RT-PCR, we found that ZFY, EIF2S3Y, TSPY, UTY, DDX3Y, USP9Y, and RBMY, but not UBA1Y, had high deduced amino acid sequence similarity and similar expression patterns with other rodents, suggesting that these genes were functional. Based on FISH and quantitative real-time PCR, all of the genes except for UTY and DDX3Y were amplified on the X and Y chromosomes with approximately 10-66 copies in the male genome. In a comparative analysis of the 372.4-kb BAC sequence and Y-linked gene transcripts from T. muenninki with the mouse Y genomic sequence, we observed that multiple-copy genes in the ancestral Y genome were nonfunctional, indicating that the gene functions were assumed by amplified copies. We also found a LTR sequence at the distal end of a SRY duplication unit, suggesting that unequal sister chromatid exchange mediated by retrotransposable elements could have been involved in SRY amplification. Our results revealed that the Y-linked genes were rescued from degeneration via translocations to other sex chromosomal regions and amplification events in T. muenninki.
• Molecular mechanism of male differentiation is conserved in the SRY-absent mammal, Tokudaia osimensis

  Tomofumi Otake, Asato Kuroiwa

  SCIENTIFIC REPORTS 6 32874  2016年09月 [査読有り][通常論文]
   

  The sex-determining gene SRY induces SOX9 expression in the testes of eutherian mammals via two pathways. SRY binds to testis-specific enhancer of Sox9 (TESCO) with SF1 to activate SOX9 transcription. SRY also up-regulates ER71 expression, and ER71 activates Sox9 transcription. After the initiation of testis differentiation, SOX9 enhances Amh expression by binding to its promoter with SF1. SOX8, SOX9 and SOX10, members of the SOXE gene family, also enhance the activities of the Amh promoter and TESCO. In this study, we investigated the regulation of these sexual differentiation genes in Tokudaia osimensis, which lacks a Y chromosome and the SRY gene. The activity of the AMH promoter was stimulated by SOXE genes and SF1. Mutant AMH promoters, with mutations in its SOX and SF1 binding sites, did not show significant activity by SOX9 and SF1. These results indicate that AMH expression was regulated by the binding of SOX9 and SF1. By contrast, SOXE genes could not enhance TESCO activity. These results indicate that TESCO enhancer activity was lost in this species. Furthermore, the activity of the SOX9 promoter was enhanced by ER71, indicating that ER71 may play an important role in the testis-specific expression of SOX9.
• Inactivation of Sonic Hedgehog Signaling and Polydactyly in Limbs of Hereditary Multiple Malformation, a Novel Type of Talpid Mutant.

  Matsubara Y, Nakano M, Kawamura K, Tsudzuki M, Funahashi JI, Agata K, Matsuda Y, Kuroiwa A, Suzuki T

  Frontiers in cell and developmental biology 4 149  2016年 [査読有り][通常論文]
  
• 特集II-6 消えるのかY染色体ーY染色体を失った哺乳類

  黒岩麻里

  生物の科学 遺伝 70 5 409 - 414 2016年 [査読有り][招待有り]
  
• How to loss the Y chromosome in Y-absent mammals

  Asato Kuroiwa

  GENES & GENETIC SYSTEMS 90 6 375 - 375 2015年12月 [査読有り][通常論文]
  
• Initiation of recombination suppression and PAR formation during the early stages of neo-sex chromosome differentiation in the Okinawa spiny rat, Tokudaia muenninki

  Chie Murata, Yoko Kuroki, Issei Imoto, Masaru Tsukahara, Naoto Ikejiri, Asato Kuroiwa

  BMC EVOLUTIONARY BIOLOGY 15 234  2015年10月 [査読有り][通常論文]
   

  Background: Sex chromosomes of extant eutherian species are too ancient to reveal the process that initiated sex-chromosome differentiation. By contrast, the neo-sex chromosomes generated by sex-autosome fusions of recent origin in Tokudaia muenninki are expected to be evolutionarily 'young', and therefore provide a good model in which to elucidate the early phases of eutherian sex chromosome evolution. Here we describe the genomic evolution of T. muenninki in neo-sex chromosome differentiation. Results: FISH mapping of a T. muenninki male, using 50 BAC clones as probes, revealed no chromosomal rearrangements between the neo-sex chromosomes. Substitution-direction analysis disclosed that sequence evolution toward GC-richness, which positively correlates with recombination activity, occurred in the peritelomeric regions, but not middle regions of the neo-sex chromosomes. In contrast, the sequence evolution toward AT-richness was observed in those pericentromeric regions. Furthermore, we showed genetic differentiation between the pericentromeric regions as well as an accelerated rate of evolution in the neo-Y region through the detection of male-specific substitutions by gene sequencing in multiple males and females, and each neo-sex-derived BAC sequencing. Conclusions: Our results suggest that recombination has been suppressed in the pericentromeric region of neo-sex chromosomes without chromosome rearrangement, whereas high levels of recombination activity is limited in the peritelomeric region of almost undifferentiated neo-sex chromosomes. We conclude that PAR might have been formed on the peritelomeric region of sex chromosomes as an independent event from spread of recombination suppression during the early stages of sex chromosome differentiation.
• Early stage of eutherian sex chromosome differentiation - Formation of recombination suppression region and high recombination region-

  Chie Murata, Yoko Kuroki, Issei Imoto, Fumio Yamada, Takamichi Jogahara, Katsushi Nakata, Asato Kuroiwa

  GENES & GENETIC SYSTEMS 89 6 280 - 280 2014年12月 [査読有り][通常論文]
  
• Analysis of cHEMGN involved in chicken-specific sex-determining mechanisms

  Tomohiro Nakata, Nana Aduma, Hiroe Izumi, Asato Kuroiwa

  GENES & GENETIC SYSTEMS 89 6 288 - 288 2014年12月 [査読有り][招待有り]
  
• Mutations in the Testis-Specific Enhancer of SOX9 in the SRY Independent Sex-Determining Mechanism in the Genus Tokudaia

  Ryutaro Kimura, Chie Murata, Yoko Kuroki, Asato Kuroiwa

  PLOS ONE 9 9 e108779  2014年09月 [査読有り][通常論文]
   

  SRY (sex-determining region Y) is widely conserved in eutherian mammals as a sex-determining gene located on the Y chromosome. SRY proteins bind to the testis-specific enhancer of SOX9 (TES) with SF1 to upregulate SOX9 expression in undifferentiated gonads of XY embryos of humans and mice. The core region within TES, named TESCO, is an important enhancer for mammalian sex determination. We show that TESCO of the genus Tokudaia lost enhancer activity caused by mutations in its SRY and SF1 binding sites. Two species of Tokudaia do not have the Y chromosome or SRY, and one species has multiple SRYs located on the neo-Y chromosome consisting of the Y fused with an autosome. The sequence of Tokudaia TESCO exhibited more than 83% identity with mouse TESCO, however, nucleotide substitution(s) were found in two out of three SRY binding sites and in five out of six SF1 binding sites. TESCO of all species showed low enhancer activity in cells co-transfected with SRY and SF1, and SOX9 and SF1 in reporter gene assays. Mutated TESCO, in which nucleotide substitutions found in SRY and SF1 binding sites were replaced with mouse sequence, recovered the activity. Furthermore, SRYs of the SRY-positive species could not activate the mutated TESCO or mouse TESCO, suggesting that SRYs lost function as a sex-determining gene any more. Our results indicate that the SRY dependent sex-determining mechanism was lost in a common ancestor of the genus Tokudaia caused by nucleotide substitutions in SRY and SF1 binding sites after emergence of a new sex-determining gene. We present the first evidence for an intermediate stage of the switchover from SRY to a new sex-determining gene in the evolution of mammalian sex-determining mechanism.
• Over-expression of DMRT1 induces the male pathway in embryonic chicken gonads

  Luke S. Lambeth, Christopher S. Raymond, Kelly N. Roeszler, Asato Kuroiwa, Tomohiro Nakata, David Zarkower, Craig A. Smith

  DEVELOPMENTAL BIOLOGY 389 2 160 - 172 2014年05月 [査読有り][通常論文]
   

  DMRT1 encodes a conserved transcription factor with an essential role in gonadal function. In the chicken, DMRT1 in located on the Z sex chromosome and is currently the best candidate master regulator of avian gonadal sex differentiation. We previously showed that knockdown of DMRT1 expression during the period of sexual differentiation induces feminisation of male embryonic chicken gonads. This gene is therefore necessary for proper testis development in the chicken. However, whether it is sufficient to induce testicular differentiation has remained unresolved. We show here that over-expression of DMRT1 induces male pathway genes and antagonises the female pathway in embryonic chicken gonads. Ectopic DMRT1 expression in female gonads induces localised SOX9 and AMH expression. It also induces expression of the recently identified Z-linked male factor, Hemogen (HEMGN). Masculinised gonads show evidence of cord-like structures and retarded female-type cortical development. Furthermore, expression of the critical feminising enzyme, aromatase, is reduced in the presence of over-expressed DMRT1. These data indicate that DMRT1 is an essential sex-linked regulator of gonadal differentiation in avians, and that it likely acts via a dosage mechanism established through the lack of global Z dosage compensation in birds. (C) 2014 Elsevier Inc. All rights reserved.
• トゲネズミーYなくしてオスがうまれる

  黒岩 麻里

  生物工学会誌 92 11 630 - 631 2014年 [査読有り][招待有り]
  
• Y染色体の運命やいかに―Yはどこまで遺伝子の数を減らせるか

  黒岩 麻里

  科学 84 7 768 - 770 岩波書店 2014年 [査読無し][招待有り]
  
• Construction of Chromosome Markers from the Lake Victoria Cichlid Paralabidochromis chilotes and Their Application to Comparative Mapping

  A. Kuroiwa, Y. Terai, N. Kobayashi, K. Yoshida, M. Suzuki, A. Nakanishi, Y. Matsuda, M. Watanabe, N. Okada

  CYTOGENETIC AND GENOME RESEARCH 142 2 112 - 120 2014年 [査読有り][通常論文]
   

  Cichlid fishes in the African Great Lakes are known as a spectacular example of adaptive radiation in vertebrates. Four linkage maps have been constructed to identify the genes responsible for adaptation and speciation, and the genetic linkages of those genes are assumed to play an important role during adaptive evolution. However, it is difficult to analyze such linkages because the linkage groups of one species do not match well with those of the other species. Chromosome markers are a powerful tool for the direct identification of linkage homology between different species. We used information about the linkage map of the Lake Malawi cichlid (Labeotropheus fuelleborni/Metriaclima zebra) to isolate bacterial artificial chromosome (BAC) clones from the BAC library of Paralabidochromis chilotes, Lake Victoria. We identified 18 of 22 P. chilotes chromosomes by single- and multi-color BAC fluorescence in situ hybridization using 19 BAC clones. Comparative mapping with the chromosome markers of P. chilotes in Astatotilapia burtoni (2n = 40) from Lake Tanganyika revealed the chromosome rearrangements that have occurred in this lineage. These chromosome markers will be useful for delineating the process of genome and chromosome evolution in African species. (C) 2013 S. Karger AG, Basel
• Functional analysis of cHEMGN involved in chicken-specific mechanisms of sex determination

  Tomohiro Nakata, Nana Aduma, Hiroe Izumi, Asato Kuroiwa

  GENES & GENETIC SYSTEMS 88 6 364 - 364 2013年12月 [査読有り][通常論文]
  
• Chicken hemogen homolog is involved in the chicken-specific sex-determining mechanism

  Tomohiro Nakata, Manabu Ishiguro, Nana Aduma, Hiroe Izumi, Asato Kuroiwa

  Proceedings of the National Academy of Sciences of the United States of America 110 9 3417 - 3422 2013年02月26日 [査読有り][通常論文]
   

  Using a comprehensive transcriptome analysis, a Z chromosomelinked chicken homolog of hemogen (cHEMGN) was identified and shown to be specifically involved in testis differentiation in early chicken embryos. Hemogen [Hemgn in mice, EDAG (erythroid differentiation-associated gene protein) in humans] was recently characterized as a hematopoietic tissue-specific gene encoding a transcription factor that regulates the proliferation and differentiation of hematopoietic cells in mammals. In chicken, cHEMGN was expressed not only in hematopoietic tissues but also in the early embryonic gonad of male chickens. The male-specific expression was identified in the nucleus of (pre)Sertoli cells after the sex determination period and before the expression of SOX9 (SRY-box 9). The expression of cHEMGN was induced in ZW embryonic gonads that were masculinized by aromatase inhibitor treatment. ZW embryos overexpressing cHEMGN, generated by infection with retrovirus carrying cHEMGN, showed masculinized gonads. These findings suggest that cHEMGN is a transcription factor specifically involved in chicken sex determination.
• 小集団化と遺伝的多様性を消失したオキナワトゲネズミ -希少種トゲネズミ 3種の生息調査と遺伝的多様性分析から

  木戸 文香, 城ヶ原 貴通, 黒岩 麻里, 越本 知大, 望月 春佳, 中家 雅隆, 村田 知慧, 三谷 匡, 山田 文雄

  霊長類研究 Supplement 29 0 232 - 232 日本霊長類学会 2013年 

   トゲネズミ属Tokudaiaは沖縄島，徳之島及び奄美大島のみに生息する我が国の固有属で，島ごとに独立種を形成し，国指定天然記念物と環境省 RDB絶滅危惧Ⅰ類に指定されている．しかし，その生息状況把握を含め，具体的保全策は図られていないのが現状である．そこで，本属 3種の生息状況と遺伝的多様性について検討した．本属 3種の生息状況は島により異なるが，とくにオキナワトゲネズミT. muenninkiは沖縄島北部（やんばる）の森林の数平方 kmの範囲にしか生息が確認されなかった．また，トクノシマトゲネズミT. tokunoshimensisはこれまで知られていた生息地点においても生息がほとんど確認されず，特に島南部での生息情報は少なかった．一方，アマミトゲネズミT. osimensisはここ数年生息数が回復し安定した生息状況になってきており，マングース防除事業による捕食圧低下の効果と考えられる．更にこれらを対象に実施した mtDNA多型解析の結果，オキナワトゲネズミでは 1個のハプロタイプしか認められなかった一方で，アマミトゲネズミでは 12個のハプロタイプが検出された．しかし，アマミトゲネズミにおけるハプロタイプのネットワーク樹分析によると，中間型のハプロタイプが多数消失していることや，特定のハプロタイプの検出頻度が多いことから，過去に個体数減少を経験したが現在は回復傾向にあることが示唆された．さらに，核 DNA分析において，オキナワトゲネズミでは多様性が低く，アマミトゲネズミでは比較的高いことが示された．これらの結果から，オキナワトゲネズミは集団サイズが小さく，遺伝的多様性を消失しており，環境変動に十分適応できない可能性が高いため，早急の保護対策が必要と考えられる．また，トクノシマトゲネズミにおいても，生息数減少や遺伝的多様性低下を起こしている可能性が高く，同様の対策が求められる．
• Y染色体は本当になくなる？

  黒岩 麻里

  細胞工学 32 2 170 - 171 2013年 [査読有り][招待有り]
  
• XIST導入とダウン症治療ー染色体治療最前線ー

  黒岩 麻里

  実験医学 31 19 3088  2013年 [査読有り][招待有り]
  
• Inhibition of the binding of MSG-intermolt-specific complex, MIC, to the sericin-1 gene promoter and sericin-1 gene expression by POU-M1/SGF-3

  Mai Kimoto, Tsuyuki Kitagawa, Isao Kobayashi, Tomohiro Nakata, Asato Kuroiwa, Shigeharu Takiya

  DEVELOPMENT GENES AND EVOLUTION 222 6 351 - 359 2012年11月 [査読有り][通常論文]
   

  The sericin-1 gene encoding a glue protein is expressed in the middle silk gland (MSG) of the silkworm, Bombyx mori. A member of the class III POU domain transcription factors, POU-M1, was cloned as the factor bound to the SC site of the sericin-1 promoter and has been proposed to be a positive transcription factor. In this study, we analyzed the expression pattern of the POU-M1 gene in fourth and fifth instars in comparison with the pattern of the sericin-1 gene. The POU-M1 gene was expressed strongly in the region anterior to the sericin-1-expressing portion of the silk gland at both feeding stages. As the sericin-1-expressing region expands from the posterior to middle portions of the MSG in the fifth instar, the POU-M1-expressing region retreated from the middle to anterior portion. Introduction of the expression vector of POU-M1 into the silk glands by gene gun technology repressed promoter activity of the sericin-1 gene, suggesting that POU-M1 regulates the sericin-1 gene negatively. An in vitro binding assay showed that POU-M1 bound not only to the SC site but also to other promoter elements newly detected in vivo. Another spatiotemporal specific factor MIC binds to these elements, and POU-M1 competed with MIC to bind at the -70 site essential for promoter activity. These results suggest that POU-M1 is involved in restricting the anterior boundary of the sericin-1-expressing region in the silk gland by inhibiting the binding of the transcriptional activator to the promoter elements.
• Sex-determining mechanism of the Y-absent ammal

  Asato Kuroiwa

  Seikagaku 84 11 931 - 934 2012年 [査読有り][招待有り]
  
• Y染色体をもたない哺乳類の性決定メカニズム

  黒岩 麻里

  生化学 84 11 931 - 934 2012年 [査読有り][招待有り]
  
• The Y chromosome of the Okinawa spiny rat, Tokudaia muenninki, was rescued through fusion with an autosome

  Chie Murata, Fumio Yamada, Norihiro Kawauchi, Yoichi Matsuda, Asato Kuroiwa

  CHROMOSOME RESEARCH 20 1 111 - 125 2012年01月 [査読有り][通常論文]
   

  The genus Tokudaia comprises three species, two of which have lost their Y chromosome and have an XO/XO sex chromosome constitution. Although Tokudaia muenninki (Okinawa spiny rat) retains the Y chromosome, both sex chromosomes are unusually large. We conducted a molecular cytogenetic analysis to characterize the sex chromosomes of T. muenninki. Using cross-species fluorescence in situ hybridization (Zoo-FISH), we found that both short arms of the T. muenninki sex chromosomes were painted by probes from mouse chromosomes 11 and 16. Comparative genomic hybridization analysis was unable to detect sex-specific regions in the sex chromosomes because both sex probes highlighted the large heterochromatic blocks on the Y chromosome as well as five autosomal pairs. We then performed comparative FISH mapping using 29 mouse complementary DNA (cDNA) clones of the 22 X-linked genes and the seven genes linked to mouse chromosome 11 (whose homologue had fused to the sex chromosomes), and FISH mapping using two T. muenninki cDNA clones of the Y-linked genes. This analysis revealed that the ancestral gene order on the long arm of the X chromosome and the centromeric region of the short arm of the Y chromosome were conserved. Whereas six of the mouse chromosome 11 genes were also mapped to Xp and Yp, in addition, one gene, CBX2, was also mapped to Xp, Yp, and chromosome 14 in T. muenninki. CBX2 is the candidate gene for the novel sex determination system in the two other species of Tokudaia, which lack a Y chromosome and SRY gene. Overall, these results indicated that the Y chromosome of T. muenninki avoided a loss event, which occurred in an ancestral lineage of T. osimensis and T. tokunoshimensis, through fusion with an autosome. Despite retaining the Y chromosome, sex determination in T. muenninki might not follow the usual mammalian pattern and deserves further investigation.
• B Chromosomes Have a Functional Effect on Female Sex Determination in Lake Victoria Cichlid Fishes

  Kohta Yoshida, Yohey Terai, Shinji Mizoiri, Mitsuto Aibara, Hidenori Nishihara, Masakatsu Watanabe, Asato Kuroiwa, Hirohisa Hirai, Yuriko Hirai, Yoichi Matsuda, Norihiro Okada

  PLOS GENETICS 7 8 e1002203  2011年08月 [査読有り][通常論文]
   

  The endemic cichlid fishes in Lake Victoria are a model system for speciation through adaptive radiation. Although the evolution of the sex-determination system may also play a role in speciation, little is known about the sex-determination system of Lake Victoria cichlids. To understand the evolution of the sex-determination system in these fish, we performed cytogenetic analysis in 11 cichlid species from Lake Victoria. B chromosomes, which are present in addition to standard chromosomes, were found at a high prevalence rate (85%) in these cichlids. In one species, B chromosomes were female-specific. Cross-breeding using females with and without the B chromosomes demonstrated that the presence of the B chromosomes leads to a female-biased sex ratio in this species. Although B chromosomes were believed to be selfish genetic elements with little effect on phenotype and to lack protein-coding genes, the present study provides evidence that B chromosomes have a functional effect on female sex determination. FISH analysis using a BAC clone containing B chromosome DNA suggested that the B chromosomes are derived from sex chromosomes. Determination of the nucleotide sequences of this clone (104.5 kb) revealed the presence of several protein-coding genes in the B chromosome, suggesting that B chromosomes have the potential to contain functional genes. Because some sex chromosomes in amphibians and arthropods are thought to be derived from B chromosomes, the B chromosomes in Lake Victoria cichlids may represent an evolutionary transition toward the generation of sex chromosomes.
• B Chromosomes Have a Functional Effect on Female Sex Determination in Lake Victoria Cichlid Fishes

  Kohta Yoshida, Yohey Terai, Shinji Mizoiri, Mitsuto Aibara, Hidenori Nishihara, Masakatsu Watanabe, Asato Kuroiwa, Hirohisa Hirai, Yuriko Hirai, Yoichi Matsuda, Norihiro Okada

  PLOS GENETICS 7 8 e1002203  2011年08月 [査読有り][通常論文]
   

  The endemic cichlid fishes in Lake Victoria are a model system for speciation through adaptive radiation. Although the evolution of the sex-determination system may also play a role in speciation, little is known about the sex-determination system of Lake Victoria cichlids. To understand the evolution of the sex-determination system in these fish, we performed cytogenetic analysis in 11 cichlid species from Lake Victoria. B chromosomes, which are present in addition to standard chromosomes, were found at a high prevalence rate (85%) in these cichlids. In one species, B chromosomes were female-specific. Cross-breeding using females with and without the B chromosomes demonstrated that the presence of the B chromosomes leads to a female-biased sex ratio in this species. Although B chromosomes were believed to be selfish genetic elements with little effect on phenotype and to lack protein-coding genes, the present study provides evidence that B chromosomes have a functional effect on female sex determination. FISH analysis using a BAC clone containing B chromosome DNA suggested that the B chromosomes are derived from sex chromosomes. Determination of the nucleotide sequences of this clone (104.5 kb) revealed the presence of several protein-coding genes in the B chromosome, suggesting that B chromosomes have the potential to contain functional genes. Because some sex chromosomes in amphibians and arthropods are thought to be derived from B chromosomes, the B chromosomes in Lake Victoria cichlids may represent an evolutionary transition toward the generation of sex chromosomes.
• Additional copies of CBX2 in the genomes of males of mammals lacking SRY, the Amami spiny rat (Tokudaia osimensis) and the Tokunoshima spiny rat (Tokudaia tokunoshimensis)

  Asato Kuroiwa, Sanae Handa, Chigusa Nishiyama, Eriko Chiba, Fumio Yamada, Shintaro Abe, Yoichi Matsuda

  CHROMOSOME RESEARCH 19 5 635 - 644 2011年07月 [査読有り][通常論文]
   

  Tokudaia osimensis (the Amami spiny rat) and Tokudaia tokunoshimensis (the Tokunoshima spiny rat) have a sex chromosome composition of XO/XO, no Y chromosome. The mammalian sex-determining gene, SRY, is also absent in these species, which indicates that these spiny rats exhibit a novel sex-determining mechanism that is independent of SRY. To identify a candidate gene that controls this mechanism, the copy numbers and chromosomal locations of 10 genes with important functions in gonadal differentiation were determined: ATRX, CBX2 (M33), DMRT1, FGF9, NR0B1 (DAX1), NR5A1 (Ad4BP/SF1), RSPO1, SOX9, WNT4, and WT1. Multiple bands were detected for NR0B1 in Southern blot analysis, which suggested the presence of multiple copies of the gene in the genomes of these two species. CBX2 was localized to two loci in both sexes of the two species by fluorescence in situ hybridization mapping: 3q24 and 6p11.2 in T. osimensis and 10q25-q26 and 14q12-q13.1 in T. tokunoshimensis. Quantification of copy numbers in the two species by quantitative real-time PCR indicated that there were two or three more copies of CBX2 per haploid genome in males (T. osimensis, n = 3; T. tokunoshimensis, n = 2) than in females (T. osimensis, n = 4; T. tokunoshimensis, n = 2), whereas NR0B1 was present as a single copy in both. The results suggest that additional copies of CBX2 in males might be involved in a novel sex-determining mechanism in species that lack SRY.
• A primer set to determine sex in the small Indian mongoose, Herpestes auropunctatus

  C. Murata, G. Ogura, A. Kuroiwa

  MOLECULAR ECOLOGY RESOURCES 11 2 386 - 388 2011年03月 [査読有り][通常論文]
   

  To enable the accurate sexing of individuals of introduced populations of the small Indian mongoose, Herpestes auropunctatus, we designed a primer set for the amplification of the sex-specific fragments EIF2S3Y and EIF2S3X. Using this primer set, the expected amplification products were obtained for all samples of genomic DNA tested: males yielded two bands and females a single band. Sequencing of each PCR product confirmed that the 769-bp fragment amplified from DNA samples of both sexes was derived from EIF2S3X, whereas the 546-bp fragment amplified only from male DNA samples was derived from EIF2S3Y. The results indicated that this primer set is useful for sex identification in this species.
• Y染色体の運命ートゲネズミを用いたY染色体進化研究ー

  黒岩 麻里

  日本受精着床学会雑誌 28 2 319 - 323 2011年 [査読有り][招待有り]
  
• The Y chromosome evolution in genus Tokudaia -Keep or loss of the Y chromosome-

  Chie Murata, Asato Kuroiwa

  GENES & GENETIC SYSTEMS 85 6 446 - 446 2010年12月 [査読有り][通常論文]
  
• Analysis of a candidate gene involved in gonadal-differentiation of chicken

  Tomohiro Nakata, Yoji Mikami, Harunobu Yunokawa, Yoichi Matsuda, Asato Kuroiwa

  GENES & GENETIC SYSTEMS 85 6 406 - 406 2010年12月 [査読有り][通常論文]
  
• The comprehensive screening of proto-Y-linked genes in XO spiny rat

  Michi Komoto, Chie Murata, Fumio Yamada, Norihiro Kawauchi, Shintaro Abe, Asato Kuroiwa

  GENES & GENETIC SYSTEMS 85 6 413 - 413 2010年12月 [査読有り][通常論文]
  
• Rediscovery after thirty years since the last capture of the critically endangered Okinawa spiny rat Tokudaia muenninki in the northern part of Okinawa Island

  Fumio Yamada, Norihiro Kawauchi, Katsushi Nakata, Shintaro Abe, Nobuhiko Kotaka, Atsushi Takashima, Chie Murata, Asato Kuroiwa

  MAMMAL STUDY 35 4 243 - 255 2010年12月 [査読有り][通常論文]
   

  The Okinawa spiny rat, Tokudaia muenninki, is a critically endangered species endemic to the northern part of Okinawa Island and may be extinct in the wild as there have been no recent sightings of the animal in its natural habitat. We initiated the present search to determine whether the spiny rat still exists in the northern part of Okinawa Island. Sensor cameras and traps were distributed across areas in which past studies had identified the location of occurrence of spiny rats. From a total of 1,276 camera-nights and 2,096 trap-nights from 2007 to 2009, we captured 24 spiny rats; however, we were only successful in identifying spiny rats in the northernmost of the areas sampled, with no indications of the spiny rat in the more southerly areas. The area in which the spiny rats were still present was estimated to be only 1-3 km(2) and is comprised of forest dominated by Castanopsis sieboldii, Lithocarpus edulis, Distylium racemosum and Schima wallichii. The trees range in age from about 30 to more than 100 years old, and have an average height of 12 m (range 7 m-16 m). Our rediscovery of the spiny rat in 2008 comes after an interval of 30 years since the previous trapping study in 1978 and seven years since indirect survey evidence from analysis of feral cat feces 2001. Measures for conservation of the location of the spiny rats are urgently required.
• The process of a Y-loss event in an XO/XO mammal, the Ryukyu spiny rat

  Asato Kuroiwa, Yasuko Ishiguchi, Fumio Yamada, Abe Shintaro, Yoichi Matsuda

  CHROMOSOMA 119 5 519 - 526 2010年10月 [査読有り][通常論文]
   

  The Ryukyu spiny rat, Tokudaia osimensis, has an XO/XO sex chromosome constitution, lacking a Y chromosome and the mammalian sex-determining gene SRY. To investigate the Y-loss event, we traced three proto-Y-linked genes, RBMY1A1, EIF2S3Y, and KDM5D, in the genome. The original Y-linked RBMY1A1 was lost as well as SRY, and the remaining RBMY1A1 was a processed pseudogene on autosome. In contrast, EIF2S3Y and KDM5D were conserved in genomes of both sexes as a result of their translocation from the Y chromosome to the X chromosome and/or autosomes. Furthermore, these genes were expressed in gonads and brains of both sexes. Our study indicated a loss of Y-linked genes with important male functions to be necessary for the Y chromosome to disappear. These functions might have been retained through the acquisition of new genes, and therefore, the Y-loss has had no harmful effect on the maintenance of this species.
• Multiple copies of SRY on the large Y chromosome of the Okinawa spiny rat, Tokudaia muenninki

  Chie Murata, Fumio Yamada, Norihiro Kawauchi, Yoichi Matsuda, Asato Kuroiwa

  CHROMOSOME RESEARCH 18 6 623 - 634 2010年09月 [査読有り][通常論文]
   

  The Okinawa spiny rat, Tokudaia muenninki, is the only species with a Y chromosome in the genus Tokudaia. Its phylogenic relationship with two XO/XO species, Tokudaia osimensis and Tokudaia tokunoshimensis, lacking a Y chromosome and the mammalian sex-determining gene SRY, is unknown. Furthermore, there has been little cytogenetic analysis of the sex chromosomes in T. muenninki. Therefore, we constructed molecular phylogenetic trees with nucleotide sequences of cyt b, RAG1, and IRBP. All trees strongly supported that T. muenninki was the first to diverge from the Tokudaia ancestor, indicating that loss of the Y chromosome and SRY occurred in the common ancestor of the two XO/XO species after T. muenninki diverged. We found that the X and Y chromosomes of T. muenninki consisted of large euchromatic and heterochromatic regions by conducting G- and C-banding analyses. PCR, Southern blotting, and FISH revealed that T. muenninki males had multiple SRY copies on the long arm of the Y chromosome. At least three of 24 SRY sequences contained a complete open reading frame (ORF). A species-specific substitution from alanine to serine was found in all copies at the DNA-binding surface within the HMG-box, suggesting that it occurred in an original SRY.
• No final answers yet on sex determination in birds

  Asato Kuroiwa

  NATURE 462 7269 34 - 34 2009年11月 [査読有り][通常論文]
  
• Y染色体を失ったほ乳類、トゲネズミ

  黒岩 麻里

  生物の科学 遺伝 63 15 - 19 2009年 [査読有り][招待有り]
  
• Centromere repositioning in the X chromosome of XO/XO mammals, Ryukyu spiny rat

  Tsuyoshi Kobayashi, Fumio Yamada, Takuma Hashimoto, Shintaro Abe, Yoichi Matsuda, Asato Kuroiwa

  CHROMOSOME RESEARCH 16 4 587 - 593 2008年06月 [査読有り][通常論文]
   

  Two species of Ryukyu spiny rat, Tokudaia osimensis and Tokudaia tokunoshimensis, have an XO/XO sex chromosome constitution with no cytogenetically visible Y chromosome in both sexes. The single X chromosomes of T. osimensis and T. tokunoshimensis are submetacentric and subtelocentric, respectively. It was therefore suggested that a pericentric inversion event occurred in the X chromosome of either species. To identify X chromosome rearrangements that have occurred between the two species, we mapped 22 mouse cDNA clones of the X-linked genes on the chromosomes of the two species by direct R-banding FISH. The gene orders of the X chromosomes were conserved in the two species, whereas the position of the centromere on the X chromosome was different. This result indicates that the rearrangement which occurred in either of the X chromosomes after the two species diverged from a common ancestor involved not pericentric inversion but centromere repositioning.
• Centromere repositioning in the X chromosome of XO/XO mammals, Ryukyu spiny rat

  Tsuyoshi Kobayashi, Fumio Yamada, Takuma Hashimoto, Shintaro Abe, Yoichi Matsuda, Asato Kuroiwa

  CHROMOSOME RESEARCH 16 4 587 - 593 2008年06月 [査読有り][通常論文]
   

  Two species of Ryukyu spiny rat, Tokudaia osimensis and Tokudaia tokunoshimensis, have an XO/XO sex chromosome constitution with no cytogenetically visible Y chromosome in both sexes. The single X chromosomes of T. osimensis and T. tokunoshimensis are submetacentric and subtelocentric, respectively. It was therefore suggested that a pericentric inversion event occurred in the X chromosome of either species. To identify X chromosome rearrangements that have occurred between the two species, we mapped 22 mouse cDNA clones of the X-linked genes on the chromosomes of the two species by direct R-banding FISH. The gene orders of the X chromosomes were conserved in the two species, whereas the position of the centromere on the X chromosome was different. This result indicates that the rearrangement which occurred in either of the X chromosomes after the two species diverged from a common ancestor involved not pericentric inversion but centromere repositioning.
• Chromosome elimination in the interspecific hybrid medaka between Oryzias latipes and O-hubbsi

  C. Sakai, F. Konno, O. Nakano, T. Iwai, T. Yokota, J. Lee, C. Nishida-Umehara, A. Kuroiwa, Y. Matsuda, M. Yamashita

  CHROMOSOME RESEARCH 15 6 697 - 709 2007年10月 [査読有り][通常論文]
   

  An interspecific hybrid medaka (rice fish) between Oryzias latipes and O. hubbsi is embryonically lethal. To gain an insight into the cellular and molecular mechanisms that cause the abnormalities occurring in the hybrid medaka, we investigated the behavior of chromosomes and the expression patterns of proteins responsible for the chromosome behavior. The number of chromosomes in the hybrid embryos gradually decreased to nearly half, since abnormal cell division with lagging chromosomes at anaphase eliminated the chromosomes from the cells. The chromosome lagging occurred at the first cleavage and continued throughout embryogenesis even after the midblastula transition. Fluorescent in-situ hybridization analyses revealed that the chromosomes derived from O. hubbsi are preferentially eliminated in both O. latipes-hubbsi and O. hubbsi-latipes embryos. Whole-mount immunocytochemical analyses using antibodies against alpha-tubulin, gamma-tubulin, inner centromere protein, Cdc20, Mad2, phospho-histone H3 and cohesin subunits (SMC1 alpha, SMC3 and Rad21) showed that the expression patterns of these proteins in the hybrid embryos are similar to those in the wild-type embryos, except for phospho-histone H3. Phospho-histone H3 present on chromosomes at metaphase was lost from normally separated chromosomes at anaphase, whereas it still existed on lagging chromosomes at anaphase, indicating that the lagging chromosomes remain in the metaphase state even when the cell has proceeded to the anaphase state. On the basis of these findings, we discuss the cellular and molecular mechanisms of chromosome elimination in the hybrid medaka.
• Comparative chromosome painting map between two Ryukyu spiny rat species, Tokudaia osimensis and Tokudaia tokunoshimensis (Muridae, Rodentia)

  Taro Nakamura, Asato Kuroiwa, Chizuko Nishida-Umehara, Kazumi Matsubara, Fumio Yamada, Yoichi Matsuda

  CHROMOSOME RESEARCH 15 6 799 - 806 2007年10月 [査読有り][通常論文]
   

  Ryukyu spiny rats (genus Tokudaia) are indigenous species that are confined to three islands of the Nansei Shoto archipelago, Amami-Oshima, Tokunoshima and Okinawa-jima, Japan. Tokudaia tokunoshimensis from Tokunoshima Island and Tokudaia osimensis from Amami-Oshima Island are closely related taxonomically, although their karyotypes are quite different: the diploid chromosome numbers and sex chromosome constitution are 2n=45, X0/X0 for T. tokunoshimensis and 2n=25, X0/X0 for T. osimensis. We conducted comparative chromosome painting with chromosome-specific DNA probes of the laboratory mouse (Mus musculus) to molecularly examine the chromosome homology between T. tokunoshimensis and T. osimensis, and deduced a possible ancestral karyotype of Tokudaia species and the process of evolutionary chromosome rearrangements. The proposed ancestral karyotype with the diploid number of 2n=48, XX/XY was similar to the karyotype of T. tokunoshimensis, and the karyotype of T. osimensis would then have been established through at least 14 chromosomal changes, mainly centric fusion and tandem fusion, from the ancestral karyotype. The close karyological relationship between the ancestral karyotypes of Tokudaia and Apodemus also suggests that the chromosomal evolution in the Tokudaia-Apodemus lineage has been very slow and has accelerated only recently in the branch leading to T. osimensis.
• Exceptional minute sex-specific region in the X0 mammal, Ryukyu spiny rat

  Tsuyoshi Kobayashi, Fumio Yamada, Takuma Hashimoto, Shintaro Abe, Yoichi Matsuda, Asato Kuroiwa

  CHROMOSOME RESEARCH 15 2 175 - 187 2007年02月 [査読有り][通常論文]
   

  The Ryukyu spiny rats (genus Tokudaia) inhabit only three islands in the Nansei Shoto archipelago in Japan, and have the variations of karyotype among the islands. The chromosome number of T. osimensis in Amami-Oshima Island is 2=25, and T. tokunoshimensis in Tokunoshima Island is 2=45, and the two species have X0 sex chromosome constitution with no cytogenetically visible Y chromosome in both sexes. We constructed the standard ideograms for these species at the 100 and 200 band levels. Comparing the banding patterns between these species, it was suggested that at least 10 times the number of Robertsonian fusions occurred in T. osimensis chromosomes. However, no karyotypic differences were observed between sexes in each species. To detect the sex-specific chromosomal region of these X0 species we applied the comparative genomic hybridization (CGH) method. Although the male- and female-derived gains and losses were detected in several chromosome regions, all of them were located in the heterochromatic and/or telomeric regions. This result suggested that the differences detected by CGH might be caused by the polymorphism on the copy numbers of repeated sequences in the heterochromatic and telomeric regions. Our result indicated that the sex-specific region, where the key to sex determination lies, is very minute in X0 species of Tokudaia.
• Chromosomal localization of mammalian Y-linked genes in X0/X0 mammal, Ryukyu spiny rat

  Yasuko Ishiguchi, Furnio Yamada, Takurna Hashimoto, Shintaro Abe, Yoichi Matsuda, Asato Kuroiwa

  CHROMOSOME RESEARCH 15 79 - 80 2007年 [査読有り][通常論文]
  
• Isolation and characterization of a novel human radiosusceptibility gene, NP95

  Masahiro Muto, Akira Fujimori, Mituru Nenoi, Kazuhiro Daino, Yoichi Matsuda, Asato Kuroiwa, Eiko Kubo, Yasuyoshi Kanari, Makoto Utsuno, Hideo Tsuji, Hideki Ukai, Kazuei Mita, Masahiko Takahagi, Kouichi Tatsumi

  RADIATION RESEARCH 166 5 723 - 733 2006年11月 [査読有り][通常論文]
   

  The murine nuclear protein Np95 has been shown to underlie resistance to ionizing radiation and other DNA insults or replication arrests in embryonic stem (ES) cells. Using the databases for expressed sequenced tags and a two-step PCR procedure, we isolated human NP95, the full-length human homologue of the murine Np95 cDNA, which consists of 4,327 bp with a single open reading frame (ORF) encoding a polypeptide of 793 amino acids and 73.3% homology to Np95. The ORF of human NP95 cDNA is identical to the UHRF1 (ubiquitin-like protein containing PHD and RING domain 1). The NP95 gene, assigned to 19p13.3, consists of 18 exons, spanning 60 kb. Several stable transformants from HEK293 and WI-38 cells that had been transfected with the antisense NP95 cDNA were, like the murine Np95-knockout ES cells, more sensitive to X rays, UV light and hydroxyurea than the corresponding parental cells. In HEK293 cells, the lack of NP95 did not affect the activities of topoisomerase Hot, whose expression had been demonstrated to be regulated by the inverted CCAAT box binding protein of 90 kDa (ICBP90) that closely resembles NP95 in amino acid sequence and in cDNA but differs greatly in genomic organization. These findings collectively indicate that the human NP95 gene is the functional orthologue of the murine Np95 gene. (c) 2006 by Radiation Research Society.
• X accumulation of LINE-1 retrotransposons in Tokudaia osimensis, a spiny rat with the karyotype XO

  LA Scott, A Kuroiwa, Y Matsuda, HA Wichman

  CYTOGENETIC AND GENOME RESEARCH 112 3-4 261 - 269 2006年 [査読有り][通常論文]
   

  The observation that LINE-1 transposable elements are enriched on the X in comparison to the autosomes led to the hypothesis that LINE-1s play a role in X chromosome inactivation. If this hypothesis is correct, loss of LINE-1 activity would be expected to result in species extinction or in an alternate pathway of dosage compensation. One such alternative pathway would be to evolve a karyotype that does not require dosage compensation between the sexes. Two of the three extant species of the Ryukyu spiny rat Tokudaia have such a karyotype; both males and females are XO. We asked whether this karyotype arose due to loss of LINE-1 activity and thus the loss of a putative component in the X inactivation pathway. Although XO Tokudaia has no need for dosage compensation, LINE-1s have been recently active in Tokudaia osimensis and show higher density on the lone X than on the autosomes.
• X accumulation of LINE-1 retrotransposons in Tokudaia osimensis, a spiny rat with the karyotype XO

  LA Scott, A Kuroiwa, Y Matsuda, HA Wichman

  CYTOGENETIC AND GENOME RESEARCH 112 3-4 261 - 269 2006年 [査読有り][通常論文]
   

  The observation that LINE-1 transposable elements are enriched on the X in comparison to the autosomes led to the hypothesis that LINE-1s play a role in X chromosome inactivation. If this hypothesis is correct, loss of LINE-1 activity would be expected to result in species extinction or in an alternate pathway of dosage compensation. One such alternative pathway would be to evolve a karyotype that does not require dosage compensation between the sexes. Two of the three extant species of the Ryukyu spiny rat Tokudaia have such a karyotype; both males and females are XO. We asked whether this karyotype arose due to loss of LINE-1 activity and thus the loss of a putative component in the X inactivation pathway. Although XO Tokudaia has no need for dosage compensation, LINE-1s have been recently active in Tokudaia osimensis and show higher density on the lone X than on the autosomes.
• Molecular cloning, expression and chromosomal localization of mouse MM-1.

  Tetsuya Inazu, Zaw Myint, Asato Kuroiwa, Yoichi Matsuda, Tamio Noguchi

  Molecular biology reports 32 4 273 - 9 2005年12月 [査読有り][通常論文]
   

  The protooncogene product Myc associates with many proteins. The isolation of the mouse MM-1; c-Myc binding protein (Myc-Modulator 1) cDNA is described. The cDNA contains a 462 bp open reading frame that encodes a polypeptide of 154 amino acid residues. The deduced amino acid sequence indicates that mouse MM-1 has a 99% identity with the sequence of human MM-1. The expression of mouse MM-1 mRNA was detected in the fetal liver, but its level was 3-fold higher than that in the normal adult liver, and was slightly increased after a partial hepatectomy. It is expressed widely in a variety of adult mouse tissues. Thus, MM-1 may play a role in liver development and growth. A bioinformatics analysis indicates that mouse MM-1 gene consists of 6 exons. Furthermore, the chromosomal location of the mouse MM-1 gene was on the F2-F3 band of chromosome 15, as determined by fluorescence in situ hybridization.
• Highly conserved linkage homology between birds and turtles: Bird and turtle chromosomes are precise counterparts of each other

  Y Matsuda, C Nishida-Umehara, H Tarui, A Kuroiwa, K Yamada, T Isobe, J Ando, A Fujiwara, Y Hirao, O Nishimura, J Ishijima, A Hayashi, T Saito, T Murakami, Y Murakami, S Kuratani, K Agata

  CHROMOSOME RESEARCH 13 6 601 - 615 2005年08月 [査読有り][通常論文]
   

  The karyotypes of birds, turtles and snakes are characterized by two distinct chromosomal components, macrochromosomes and microchromosomes. This close karyological relationship between birds and reptiles has long been a topic of speculation among cytogeneticists and evolutionary biologists; however, there is scarcely any evidence for orthology at the molecular level. To define the conserved chromosome synteny among humans, chickens and reptiles and the process of genome evolution in the amniotes, we constructed comparative cytogenetic maps of the Chinese soft-shelled turtle (Pelodiscus sinensis) and the Japanese four-striped rat snake (Elaphe quadrivirgata) using cDNA clones of reptile functional genes. Homology between the turtle and chicken chromosomes is highly conserved, with the six largest chromosomes being almost equivalent to each other. On the other hand, homology to chicken chromosomes is lower in the snake than in the turtle. Turtle chromosome 6q and snake chromosome 2p represent conserved synteny with the chicken Z chromosome. These results suggest that the avian and turtle genomes have been well conserved during the evolution of the Arcosauria. The avian and snake sex Z chromosomes were derived from different autosomes in a common ancestor, indicating that the causative genes of sex determination may be different between birds and snakes.
• Cloning, expression and chromosomal assignment of human pleckstrin 2

  T Inazu, A Kuroiwa, Y Matsuda, K Miyamoto

  MOLECULAR BIOLOGY REPORTS 32 1 35 - 40 2005年03月 [査読有り][通常論文]
   

  We report the isolation of human pleckstrin 2 cDNA. The cDNA contains a 1059 bp open reading frame encoding a polypeptide of 353 amino acid residues. The deduced amino acid sequence indicates that pleckstrin 2 contains two pleckstrin homology domains and a DEP (dishvelled, egl-10, and pleckstrin) domain and had a 95% identity with the sequence of mouse pleckstrin 2. Northern blot and a reverse transcription-coupled polymerase chain reaction analysis revealed that pleckstrin 2 mRNA is widely expressed in a variety of cell lines. The chromosomal location of the mouse pleckstrin 2 gene was on the D3 band of chromosome 12, as determined by fluorescence in situ hybridization and the human pleckstrin 2 gene was mapped to chromosome 14q24.1 by a bioinformatics analysis.
• 鳥類における遺伝子量補正機構

  黒岩麻里, 松田洋一

  蛋白質・核酸・酵素 48 14 1934 - 1939 共立出版 2003年11月 [査読有り][通常論文]
  
• CPEB2, a novel putative translational regulator in mouse haploid germ cells

  Y Kurihara, M Tokuriki, R Myojin, T Hori, A Kuroiwa, Y Matsuda, T Sakurai, M Kimura, NB Hecht, S Uesugi

  BIOLOGY OF REPRODUCTION 69 1 261 - 268 2003年07月 [査読無し][通常論文]
   

  Translational control of specific mRNAs by cytoplasmic polyadenylation has fundamental roles in gametogenesis. The cytoplasmic polyadenylation element binding (CPEB) protein regulates cytoplasmic polyadenylation of mRNAs as a trans factor in oogenesis and spermatogenesis. The CPEB protein contains two RNA recognition motifs and a Zn-finger structure. Proteins (KIAA0940 and KIAA1673) with similar structures are predicted from the genome database, but nothing is known about their expression and function. Here, we report another novel member of the CPEB protein family, CPEB2. Comparison of the amino acid sequences of CPEB family members suggests that the family can be divided structurally and, perhaps, functionally into two groups: the CPEB group, and the CPEB2-KIAA0940-KIAA1673 group. The CPEB2 maps to mouse chromosome distal 5113 and is abundantly expressed in testis. However, it was detected by reverse transcription-polymerase chain reaction in all tissues that we examined. It preferentially binds to poly(U) and localizes to the cytoplasm in transfected HeLa cells. The CPEB2 is expressed postmeiotically in mouse spermatogenesis, suggesting a possible role in translational regulation of stored mRNAs in transcriptionally inactive haploid spermatids.
• Paired activating and inhibitory immunoglobulin-like receptors, MAIR-I and MAIR-II, regulate mast cell and macrophage activation

  K Yotsumoto, Y Okoshi, K Shibuya, S Yamazaki, S Tahara-Hanaoka, S Honda, M Osawa, A Kuroiwa, Y Matsuda, DG Tenen, A Iwama, H Nakauchi, A Shibuya

  JOURNAL OF EXPERIMENTAL MEDICINE 198 2 223 - 233 2003年07月 [査読有り][通常論文]
   

  Immune responses are regulated by opposing positive and negative signals triggered by the interaction of activating and inhibitory cell surface receptors with their ligands. Here, we describe novel paired activating and inhibitory immunoglobulin-like receptors, designated myeloid-associated immunoglobulin-like receptor (MAIR) I and MAIR-II, whose extracellular domains are highly conserved by each other. MAIR-I, expressed on the majority of myeloid cells, including macrophages, granulocytes, mast cells, and dendritic cells, contains the tyrosine-based sorting motif and the immunoreceptor tyrosine-based inhibitory motif-like sequences in the cytoplasmic domain and mediates endocytosis of the receptor and inhibition of IgE-mediated degranulation from mast cells. On the other hand, MAIR-II, expressed on subsets of peritoneal macrophages and B cells, associates with the immunoreceptor tyrosine-based activation motif-bearing adaptor DAP12 and stimulates proinflammatory cytokines and chemokine secretions from macrophages. Thus, MAIR-I and MAIR-II play important regulatory roles in cell signaling and immune responses.
• Molecular cloning of mammalian Spred-3 which suppresses tyrosine kinase-mediated Erk activation

  R Kato, A Nonami, T Taketomi, T Wakioka, A Kuroiwa, Y Matsuda, A Yoshimura

  BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 302 4 767 - 772 2003年03月 [査読無し][通常論文]
   

  We have reported on Spred-1 and Spred-2, which inhibit MAP kinase activation by interacting with c-kit and ras/raf.. Here, we report the cloning of a third member in this family, Spred-3. Spred-3 is expressed exclusively in the brain and its gene locates in chromosome 19q13.13 in human. Like Spred-1 and -2, Spred-3 contains an EVH1 domain in the N-terminus and a Sprouty-related cysteine-rich region (SPR domain) in the C-terminus that is necessary for membrane localization. However, Spred-3 does not possess a functional c-kit binding domain (KBD), since the critical amino acid Arg residue in this region was replaced with Gly in Spred-3. Although Spred-3 suppressed growth factor-induced MAP kinase (Erk) activation, inhibitory activity of Spred-3 was lower than that of Spred-1 or Spred-2. By the analysis of chimeric molecules between Spred-3 and Spred-1, we found that the SPR domain, rather than KBD, is responsible for efficient Erk suppression. The finding of Spred-3 revealed the presence of a novel family of regulators for the Ras/MAP kinase pathway, each member of which may have different specificities for extracellular signals. (C) 2003 Elsevier Science (USA). All rights reserved.
• Identification of chromosome rearrangements between the laboratory mouse (Mus musculus) and the Indian spiny mouse (Mus platythrix) by comparative FISH analysis

  K Matsubara, C Nishida-Umehara, A Kuroiwa, K Tsuchiya, Y Matsuda

  CHROMOSOME RESEARCH 11 1 57 - 64 2003年01月 [査読無し][通常論文]
   

  Comparative chromosome painting was applied to the Indian spiny mouse (Mus platythrix) with mouse (M. musculus) chromosome-specific probes for understanding the process of chromosome rearrangements between the two species. The chromosome locations of the 5S and 18S-28S ribosomal RNA genes and the order of the 119 and Tcp-1 genes in the In(17)2 region of the t-complex were also compared. All the painting probes were successfully hybridized to the Indian spiny mouse chromosomes, and a total of 27 segments homologous to mouse chromosomes were identified. The comparative FISH analysis revealed that tandem fusions were major events in the chromosome evolution of the Indian spiny mouse. In addition, other types of chromosome rearrangements, i.e. reciprocal translocations and insertions, were also included.
• FISH analysis of 142 EGFP transgene integration sites into the mouse genome

  T Nakanishi, A Kuroiwa, S Yamada, A Isotani, A Yamashita, A Tairaka, T Hayashi, T Takagi, M Ikawa, Y Matsuda, M Okabe

  GENOMICS 80 6 564 - 574 2002年12月 [査読無し][通常論文]
   

  Production of transgenic animals is an important technique for studying various biological processes. However, whether the integration of a particular transgene occurs randomly in the mouse genome has not been determined. Analysis by fluorescence in situ hybridization of the integration sites of the 142 EGFP (a mutant of green fluorescent protein) transgenic lines that we produced showed that the transgenes had become incorporated into every mouse chromosome. A single integration site was observed in 82.4% of the lines. The concomitant integrations of transgene into two different loci were observed in 15 cases (10.6%). In 3 cases, the transgenic founder mice showed chimerism in integration sites (2.1%). Chromosomal translocation was observed in 7 cases (4.9%). Moreover, when we statistically analyzed the transgene integration sites of these mouse lines, they were shown to distribute unevenly throughout the genome. This is the first report to analyze the transgene integration sites by producing more than 100 transgenic mouse lines.
• FISH analysis of 142 EGFP transgene integration sites into the mouse genome

  T Nakanishi, A Kuroiwa, S Yamada, A Isotani, A Yamashita, A Tairaka, T Hayashi, T Takagi, M Ikawa, Y Matsuda, M Okabe

  GENOMICS 80 6 564 - 574 2002年12月 [査読有り][通常論文]
   

  Production of transgenic animals is an important technique for studying various biological processes. However, whether the integration of a particular transgene occurs randomly in the mouse genome has not been determined. Analysis by fluorescence in situ hybridization of the integration sites of the 142 EGFP (a mutant of green fluorescent protein) transgenic lines that we produced showed that the transgenes had become incorporated into every mouse chromosome. A single integration site was observed in 82.4% of the lines. The concomitant integrations of transgene into two different loci were observed in 15 cases (10.6%). In 3 cases, the transgenic founder mice showed chimerism in integration sites (2.1%). Chromosomal translocation was observed in 7 cases (4.9%). Moreover, when we statistically analyzed the transgene integration sites of these mouse lines, they were shown to distribute unevenly throughout the genome. This is the first report to analyze the transgene integration sites by producing more than 100 transgenic mouse lines.
• Molecular and cytogenetic characterization of the mouse ATP-binding cassette transporter Abcg4

  M Yoshikawa, H Yabuuchi, A Kuroiwa, Y Ikegami, Y Sai, Tamai, I, A Tsuji, Y Matsuda, H Yoshida, T Ishikawa

  GENE 293 1-2 67 - 75 2002年06月 [査読無し][通常論文]
   

  We have cloned a new mouse ATP-binding cassette (ABC) transporter, Abcg4, from a complementary DNA (cDNA) library of mouse brain. The cloned Abcg4 cDNA encodes a protein consisting of 646 amino acids and including one ATP-binding cassette and six transmembrane domains. The Abcg4 protein exhibits high identity (96%) with human ABCG4 in terms of the amino acid sequence. Fluorescence in situ hybridization with mouse and rat chromosomes has revealed that the Abcg4 gene is located on chromosomes 9A5.3 and 8q22 distal in mouse and rat, respectively. In these loci on mouse and rat chromosomes, conserved linkage homologies were hitherto identified with human chromosome 11q23, which involves the human ABCG4 gene. The mouse Abcg4 gene as well as the human ABCG4 gene each has a total of 14 exons to encode its respective protein. High transcript levels of mouse Abcg4 were detected in mouse brain, spleen, eye, and bone marrow. Taken together, our data on the chromosomal location, gene homology, protein structure, and phylogenetic relationships strongly support the idea that mouse Abcg4 is orthologue to the human ABCG4. By functionally analyzing the mouse Abcg4 protein, we may better understand the biological role of the human ABCG4 transporter. (C) 2002 Published by Elsevier Science B.V.
• The ADAM1a and ADAM1b genes, instead of the ADAM1 (fertilin α) gene, are localized on mouse chromosome 5.

  NISHIMURA H, KIM E, FUJIMORI T, KASHIWABARA S, KUROIWA A, MATSUDA Y, BABA T

  Gene 291 1-2 67 - 76 2002年05月 [査読無し][通常論文]
   

  Fertilin is reported to be a heterodimeric protein composed of (A) under bar (D) under bar isintegrin (A) under bar nd Motalloprotease 1 (ADAM1, fertilin alpha) and ADAM2 (fertilin P) located on the sperm surface. In the process of clarifying the molecular basis of mouse ADAM1, we have identified two intronless mouse genes encoding different isoforms of ADAM1, termed ADAM1a and ADAM1b. The amino acid sequences of ADAM1a and ADAM1b deduced from the DNA sequences were homologous to each other (99% identity) in the pro- and metalloprotease domains, whereas the C-terminal half region of ADAM la, including the disintegrin and Cys-rich domains, shared only a low degree of identity (37%) with that of ADAM1b. These two genes were both localized on mouse chromosome 5 as a single copy gene, and were expressed specifically in the testis. These data demonstrate the presence of the ADAM1a (Adam1a) and ADAM1b (Adatnlb) genes in mouse, instead of the ADAM1 gene, and may imply different roles of ADAM1a and ADAM1b in spermatogenesis, sperm maturation, and/or fertilization. (C) 2002 Elsevier Science B.V. All rights reserved.
• The ADAM1a and ADAM1b genes, instead of the ADAM1 (fertilin alpha) gene, are localized on mouse chromosome 5

  H Nishimura, E Kim, T Fujimori, S Kashiwabara, A Kuroiwa, Y Matsuda, T Baba

  GENE 291 1-2 67 - 76 2002年05月 [査読有り][通常論文]
   

  Fertilin is reported to be a heterodimeric protein composed of (A) under bar (D) under bar isintegrin (A) under bar nd Motalloprotease 1 (ADAM1, fertilin alpha) and ADAM2 (fertilin P) located on the sperm surface. In the process of clarifying the molecular basis of mouse ADAM1, we have identified two intronless mouse genes encoding different isoforms of ADAM1, termed ADAM1a and ADAM1b. The amino acid sequences of ADAM1a and ADAM1b deduced from the DNA sequences were homologous to each other (99% identity) in the pro- and metalloprotease domains, whereas the C-terminal half region of ADAM la, including the disintegrin and Cys-rich domains, shared only a low degree of identity (37%) with that of ADAM1b. These two genes were both localized on mouse chromosome 5 as a single copy gene, and were expressed specifically in the testis. These data demonstrate the presence of the ADAM1a (Adam1a) and ADAM1b (Adatnlb) genes in mouse, instead of the ADAM1 gene, and may imply different roles of ADAM1a and ADAM1b in spermatogenesis, sperm maturation, and/or fertilization. (C) 2002 Elsevier Science B.V. All rights reserved.
• Biallelic expression of Z-linked genes in male chickens

  A Kuroiwa, T Yokomine, H Sasaki, M Tsudzuki, K Tanaka, T Namikawa, Y Matsuda

  CYTOGENETIC AND GENOME RESEARCH 99 1-4 310 - 314 2002年 [査読無し][通常論文]
   

  In birds, females are heterogametic (ZW), while males are homogametic (ZZ). It has been proposed that there is no dosage compensation for the expression of Z-linked genes in birds. In order to examine if the genes are inactivated on one of the two Z chromosomes, we analyzed the allelic expression of the B4GALT1 and CHD-Z genes on Z chromosomes in male chickens. One base substitution was detected among 15 chicken breeds and lines examined for each gene, and cross mating was made between the breeds or lines with polymorphism. cDNAs were synthesized from cultured cell colonies each derived from a single cell of an F1 male embryo. The allelic expression of the B4GALT1 gene was examined by restriction fragment length polymorphism analysis of the PCR products digested with RsaI, and that of the CHD-Z gene by the single nucleotide primer extension (SNuPE) method. Both of the genes displayed biallelic expression, suggesting that these Z-linked genes were not subject to inactivation in male chickens. Comparison between expression levels in males and females by real-time quantitative PCR suggested that expression was compensated for the CHD-Z gene but not for the B4GALT1 gene. Copyright (C) 2002 S. Karger AG, Basel.
• Chromosome assignment of eight SOX family genes in chicken

  A Kuroiwa, M Uchikawa, Y Kamamchi, H Kondoh, C Nishida-Umehara, J Masabanda, DK Griffin, Y Matsuda

  CYTOGENETIC AND GENOME RESEARCH 98 2-3 189 - 193 2002年 [査読有り][通常論文]
   

  Chromosome locations of the eight SOX family genes, SOX1, SOX2, SOX3, SOX5, SOX9, SOX10, SOX14 and SOX21, were determined in the chicken by fluorescence in situ hybridization. The SOX1 and SOX21 genes were localized to chicken chromosome 1q3.1 --> q3.2, SOX5 to chromosome 1p1.6 --> p1.4, SOX10 to chromosome 1p1.6, and SOX3 to chromosome 4p1.2 --> p1.1. The SOX2 and SOX14 genes were shown to be linked to chromosome 9 using two-colored FISH and chromosome painting, and the SOX9 gene was assigned to a pair of microchromosomes. These results suggest that these SOX genes form at least three clusters on chicken chromosomes. The seven SOX genes, SOX1, SOX2, SOX3, SOX5, SOX10, SOX14 and SOX21 were localized to chromosome segments with homologies to human chromosomes, indicating that the chromosome locations of SOX family genes are highly conserved between chicken and human. Copyright (C) 2002 S. Karger AG, Basel.
• Cloning, genomic structure and chromosomal localization of the gene encoding mouse DNA helicase RECQL5 beta

  T Ohhata, R Araki, R Fukumura, A Kuroiwa, Y Matsuda, M Abe

  GENE 280 1-2 59 - 66 2001年12月 [査読有り][通常論文]
   

  Five members of the RecQ helicase family, RECQL, WRN, BLM, RTS and RECQL5, have been found in human and three of them (WRN, BLM and RTS) were disclosed to be the genes responsible for Werner, Bloom and Rothmund-Thomson syndromes. respectively. RECQL5 (RecQ helicase protein-like 5) was isolated as the fifth member of the family in humans through a search of homologous expressed sequence tags. The gene is expressed with at least three alternative splicing products, alpha, beta and gamma. Here, we isolated mouse RECQL5beta and determined the DNA sequence of full-length cDNA as well as the genome organization and chromosome locus. The mouse RECQL5beta gene consists of 2949 bp coding 982 amino acid residues. Comparison of amino acid sequence among human (Homo sapiens), mouse (Mus musculus), Drosophila metanogaster and Caenorhabditis elegans RECQL5beta homologs revealed three portions of highly conserved regions in addition to the helicase domain. Nineteen exons are dispersed over 40 kbp in the genome and all of the acceptor and donor sites for the splicing of each exon conform to the GT/AG rule. The gene is localized to the mouse chromosome 11E2, which has a syntenic relation to human 17q25.2-q25.3 where human RECQL5beta exists. Our genetic characterizations of the mouse RECQL5beta gene will contribute to functional studies on the RECQL5beta products. (C) 2001 Elsevier Science B.V. All rights reserved.
• Two mouse piwi-related genes: miwi and mili

  S Kuramochi-Miyagawa, T Kimura, K Yomogida, A Kuroiwa, Y Tadokoro, Y Fujita, M Sato, Y Matsuda, T Nakano

  MECHANISMS OF DEVELOPMENT 108 1-2 121 - 133 2001年10月 [査読有り][通常論文]
   

  Genes belonging to the piwi family are required for stem cell self-renewal in diverse organisms. We cloned mouse homologues of piwi by RT-PCR using degenerative primers. The deduced amino acid sequences of mouse homologues MIWI and MILI showed that each contains a well-conserved C-terminal PIWI domain and that each shares significant homology with PIWI and their human counterparts HIWI. Both miwi and mili were found in germ cells of adult testis by in situ hybridization, suggesting that these genes may function in spermatogenesis. Furthermore, mili was expressed in primordial germ cells (PGCs) of developing mouse embryos and may therefore play a role during germ cell formation. MIWI may be involved in RNA processing or translational regulation, since MIWI was found to possess RNA binding activity. Our data suggest that miwi and mili regulate spermatogenesis and primordial germ cell production. (C) 2001 Elsevier Science Ireland Ltd. All rights reserved.
• Structure and expression of the Ah receptor repressor gene

  T Baba, J Mimura, K Gradin, A Kuroiwa, T Watanabe, Y Matsuda, J Inazawa, K Sogawa, Y Fujii-Kuriyama

  JOURNAL OF BIOLOGICAL CHEMISTRY 276 35 33101 - 33110 2001年08月 [査読有り][通常論文]
   

  The aryl hydrocarbon receptor (AhR) repressor (AhRR) gene has been isolated and characterized from a mouse genomic library. The gene is distributed as 11 exons in a total length of about 60 kilobase pairs. Fluorescence in situ hybridization analysis has shown that the AhRR gene is located at mouse chromosome 13C2, at rat chromosome 1p11.2, and at human chromosome 5p15.3. The AhRR gene has a TATA-less promoter and several transcription start sites. In addition, putative regulatory DNA sequences such as xenobiotic responsive element (XRE), GC box, and NF-kappaB-binding sites have been identified in the 5'-upstream region of the AhRR gene. Transient transfection analyses of HeLa cells with reporter genes that contain deletions and point mutations in the AhRR promoter revealed that all three XREs mediated the inducible expression of the AhRR gene by 3-methylcholanthrene treatment, and furthermore, GC box sequences were indispensable for a high level of inducible expression and for constitutive expression. Moreover, by using gel mobility shift assays we were able to show that the AhR/Arnt heterodimer binds to the XREs with very low affinity, which is due to three varied nucleotides outside the XRE core sequence. We have also shown that Sp1 and Sp3 can bind to the GC boxes. Finally, both transient transfection analysis and gel mobility shift assay revealed that the AhRR gene is up-regulated by a p65/p50 heterodimer that binds to the NF-kappaB site when the cells has been exposed to 12-O-tetradecanoylphorbol-13-acetate, and this inducible expression was further enhanced by cotreatment of 12-O-tetradecanoylphorbol-13-acetate and 3-methylcholanthrene.
• Cloning, chromosomal mapping, and characteristic 5 '-UTR sequence of murine cytosolic sialidase

  K Kotani, A Kuroiwa, T Saito, Y Matsuda, T Koda, S Kijimoto-Ochiai

  BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 286 2 250 - 258 2001年08月 [査読有り][通常論文]
   

  We have totally sequenced a cytosolic sialidase [EC 3.2.1.18] by RT-PCR from the murine thymus (murine thymic sialidase, NITS) which has a 1844-base length (encoding 385 amino acids including two sialidase motifs) and is the longest cytosolic sialidase ever reported. MTS has high and relatively low homologies with those of mammalian cytosolic sialidases from the mouse brain (99%), rat (91%), and human skeletal muscle (75%), and those of the mouse lysosomal (47%) and membrane-bound (51%) sialidases, respectively. Chromosomal mapping, being the first report of mouse cytosolic sialidase gene, showed that the MTS gene is localized to the distal part of mouse chromosome 1D and to rat chromosome 9q36. RT-PCR with the site-specific primers revealed that the coding region was expressed in all organs tested, but expressions including the 5'-UTR were barely detectable except for in the upper-thymic fraction. Also, soluble sialidase activity in the thymus was the highest of these organs. There were mRNA instability signals and AT-rich regions in 143 bp of NITS 5'-end. (C) 2001 Academic Press.
• Efficient chromosomal transposition of a Tc1/mariner-like transposon Sleeping Beauty in mice

  K Horie, A Kuroiwa, M Ikawa, M Okabe, G Kondoh, Y Matsuda, J Takeda

  PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 98 16 9191 - 9196 2001年07月 [査読有り][通常論文]
   

  The presence of mouse embryonic stem (ES) cells makes the mouse a powerful model organism for reverse genetics, gene function study through mutagenesis of specific genes. In contrast, forward genetics, identification of mutated genes responsible for specific phenotypes, has an advantage to uncover novel pathways and unknown genes because no a priori assumptions are made about the mutated genes. However, it has been hampered in mice because of the lack of a system in which a large-scale mutagenesis and subsequent isolation of mutated genes can be performed efficiently. Here, we demonstrate the efficient chromosomal transposition of a Tc1/mariner-like transposon, Sleeping Beauty, in mice. This system allows germ-line mutagenesis in vivo and will facilitate certain aspects of phenotype-driven genetic screening in mice.
• Cloning and chromosome mapping of human and chicken Iroquois (IRX) genes

  K. Ogura, K. Matsumoto, A. Kuroiwa, T. Isobe, T. Otoguro, V. Jurecic, A. Baldini, Y. Matsuda, T. Ogura

  Cytogenetic and Genome Research 92 3-4 320 - 325 2001年06月28日 

  Three highly homologous homeobox genes (<i>caupolican, araucan </i>and <i>mirror</i>) have been identified in Drosophila. These genes belong to the novel <i>Iroquois </i>complex, which acts as a pre-pattern molecule in Drosophila neurogenesis. Recently several vertebrate <i>Iroquois </i>homologues <i>(Irx) </i>were isolated and found to be involved in pattern formation of various tissues. Here we report cytogenetic mapping of four human and five chicken <i>Iroquois </i>genes by FISH. Our findings revealed that vertebrate <i>Irx </i>genes are clustered at two different loci.
• Chromosomal mapping of 18S-28S rRNA genes and 10 cDNA clones of human chromosome 1 in the musk shrew (Suncus murinus)

  A Kuroiwa, K Matsubara, T Nagase, N Nomura, JK Seong, A Ishikawa, RVP Anunciado, K Tanaka, T Yamagata, JS Masangkay, VB Dang, T Namikawa, Y Matsuda

  JOURNAL OF HEREDITY 92 3 282 - 287 2001年05月 [査読有り][通常論文]
  
• Chromosome assignment of four plexin A genes <i>(Plxna1, Plxna2, Plxna3, Plxna4)</i> in mouse, rat, Syrian hamster and Chinese hamster

  A. Kuroiwa, F. Suto, H. Fujisawa, Y. Matsuda

  Cytogenetic and Genome Research 92 1-2 127 - 129 2001年04月02日 

  We determined chromosome locations of four plexin A subfamily genes, <i>Plxna1</i>, <i>Plxna2</i>, <i>Plxna3</i> and <i>Plxna4</i>, in four rodent species, mouse, rat, Syrian hamster and Chinese hamster, by fluorescence in situ hybridization. <i>Plxna1</i>, <i>Plxna2</i>, <i>Plxna3</i> and <i>Plxna4</i> were localized to Chr 6E2, 1H6, XB-C1 and 6B1 in mouse, Chr 4q34.1, 13q26→q27, Xq37.1→q37.2 and 4q21.3→q22 in rat, Chr 8qb1.1→qb1.3, 11qb8, Xpb8 and 5qb3.3 in Syrian hamster, and Chr 8q1.2, 5q3.7, Xp2.7 and 1q2.2→q2.3 in Chinese hamster, respectively. All the mouse and rat plexin A genes were localized to chromosome regions where conserved homology has been identified among human, mouse and rat.
• Comparative FISH mapping of human cDNA clones to chromosomes of the musk shrew (Suncus murinus, Insectivora)

  K Matsubara, A Ishikawa, A Kuroiwa, T Nagase, N Nomura, T Namikawa, Y Matsuda

  CYTOGENETICS AND CELL GENETICS 93 3-4 258 - 262 2001年 [査読有り][通常論文]
   

  Forty-one cDNA clones of human functional genes were newly mapped to chromosomes of the musk shrew (Suncus murinus, Insectivora) by fluorescence in situ hybridization, and a comparative cytogenetic map of 51 genes, including 10 genes reported in our previous study, was constructed between human (HSA) and musk shrew (SMU) chromosomes. In this comparative map, the 51 genes localized to human autosomes, except HSA 8, 16, and 20, were mapped to 15 shrew autosomes, except SMU 4, 16, 17 and 18. Twelve conserved segments were identified between human and shrew chromosomes, and six segments among the musk shrew, human, and mouse. Our results defined the presence of at least one inversion and several interchromosomal rearrangements that occurred during evolution after the two species diverged from a common ancestor. Localization of three major histocompatibility complex (MHC) genes to shrew chromosome 3 suggested that the MHC genes of the musk shrew are located in a cluster on chromosome 3. The cytogenetic map constructed in this study is the first cytogenetic map with many functional genes in insectivore species. This approach provides clues for clarifying the chromosomal evolution in this order. Copyright (C) 2001 S. Karger AG, Basel.
• Sequence polymorphisms, allelic expression status and chromosome locations of the chicken IGF2 and MPR1 genes

  T Yokomine, A Kuroiwa, K Tanaka, M Tsudzuki, Y Matsuda, H Sasaki

  CYTOGENETICS AND CELL GENETICS 93 1-2 109 - 113 2001年 [査読有り][通常論文]
   

  By screening 26 chicken breeds and lines, DNA polymorphisms were identified in the IGF2 and MPR1 genes, of which mammalian homologues are parentally imprinted, and the GAPD gene, a housekeeping control. Using the polymorphisms as genetic markers, we found that all three genes are expressed biallelically in embryonic tissues. IGF2 and MPR1 were mapped on chicken chromosomes 5 and 3. respectively, by fluorescence in situ hybridization, demonstrating conserved linkage homology between mammals and birds. Copyright (C) 2001 S. Karger AG, Basel.
• Expression and chromosome location of hamster Ku70 and Ku80

  M Koike, A Kuroiwa, A Koike, T Shiomi, Y Matsuda

  CYTOGENETICS AND CELL GENETICS 93 1-2 52 - 56 2001年 [査読有り][通常論文]
   

  Ku proteins play an important role in DNA double-strand break (DSB) repair. chromosome maintenance. and growth regulation. To understand the fundamental characteristics of Ku proteins, we examined the electrophoretic mobility and expression of hamster Ku70 and Ku80 and determined the chromosome locations of their genes. The electrophoretic mobility of hamster Ku proteins are different from that of human Ku proteins. No significant changes in the quantity of Ku proteins were observed in CHO-K1 cells treated with 10 Gy of ionizing radiation. suggesting that both proteins are expressed constitutively in amounts adequate to repair DNA DSBs. The chromosome locations of the Ku genes were determined by direct R-banding fluorescence in situ hybridization. The Ku70 gene was localized to Syrian hamster chromosome 4qa4.1 --> qa4.2 and Chinese hamster chromosome 2p3.1, and the Ku80 gene was localized to Syrian hamster chromosome 4qb5 --> qb6.1 and Chinese hamster chromosome 2p3.5 --> p3.6. These results provide clues to the biological functions of Ku, as well as useful information for constructing comparative chromosome maps between hamsters and other mammalian species, including human, mouse, and rat. Copyright (C) 2001 S. Karger AG, Basel.
• Cloning and chromosome mapping of human and chicken Iroquois (IRX) genes

  K Ogura, K Matsumoto, A Kuroiwa, T Isobe, T Otoguro, Jurecic, V, A Baldini, Y Matsuda, T Ogura

  CYTOGENETICS AND CELL GENETICS 92 3-4 320 - 325 2001年 [査読有り][通常論文]
   

  Three highly homologous homeobox genes (caupolican, araucan and mirror) have been identified in Drosophila. These genes belong to the novel Iroquois complex, which acts as a pre-pattern molecule in Drosophila neurogenesis. Recently several vertebrate Iroquois homologues (Irx) were isolated and found to be involved in pattern formation of various tissues. Here we report cytogenetic mapping of four human and five chicken Iroquois genes by FISH. Our findings revealed that vertebrate Irx genes are clustered at two different loci. Copyright (C) 2001 S. Karger AG, Basel.
• Construction of comparative cytogenetic maps of the Chinese hamster to mouse, rat and human

  A Kuroiwa, K Tsuchiya, K Matsubara, T Namikawa, Y Matsuda

  CHROMOSOME RESEARCH 9 8 641 - 648 2001年 [査読無し][通常論文]
   

  We constructed comparative cytogenetic maps of the Chinese hamster to mouse, rat and human by fluorescence in-situ hybridization using 36 cDNA clones of mouse, rat, Syrian hamster, Chinese hamster and human functional genes. In this study, 30 out of the 36 genes were newly mapped to Chinese hamster chromosomes. The chromosomal homology of the Chinese hamster was identified and arranged in 19, 19 and 18 segments of conserved synteny in mouse, rat and human, respectively. Additionally, two of the 19 segments homologous to mouse chromosomes were initially identified in this study.
• Chromosome assignment of four plexin A genes (Plxna1, Plxna2, Plxna3, Plxna4) mouse, rat, Syrian hamster and Chinese hamster

  A Kuroiwa, F Suto, H Fujisawa, Y Matsuda

  CYTOGENETICS AND CELL GENETICS 92 1-2 127 - 129 2001年 [査読有り][通常論文]
   

  We determined chromosome locations of four plexin A subfamily genes, Plxna1, Plxna2, Plxna3 and Plxna4, in four rodent species, mouse, rat, Syrian hamster and Chinese hamster, by fluorescence in situ hybridization. Plxna1, Plxna2, Plxna3 and Plxna4 were localized to Chr 6E2, 1H6, XB-C1 and 6B1 in mouse, Chr 4q34.1, 13q26 --> q27, Xq37.1 --> q37.2 and 4q21.3 --> q22 in rat, Chr 8qb1.1 --> qb1.3, 11qb8, Xpb8 and 5qb3.3 in Syrian hamster, and Chr 8q1.2, 5q3.7, Xp2.7 and 1q2.2 --> q2.3 in Chinese hamster, respectively. All the mouse and rat plexin A genes were localized to chromosome regions where conserved homology has been identified among human, mouse and rat. Copyright (C) 2001 S. Karger AG, Basel.
• Conservation of the rat X chromosome gene order in rodent species

  A Kuroiwa, K Tsuchiya, T Watanabe, H Hishigaki, E Takahashi, T Namikawa, Y Matsuda

  CHROMOSOME RESEARCH 9 1 61 - 67 2001年 [査読無し][通常論文]
   

  We constructed the comparative cytogenetic maps of X chromosomes in three rodent species, Indian spiny mouse (Mus platythrix), Syrian hamster and Chinese hamster, using 26 mouse cDNA clones. Twenty-six, 22 and 22 out of the 26 genes, which were mapped to human, mouse and rat X chromosomes in our previous study, were newly localized to X chromosomes of Indian spiny mouse, and Syrian and Chinese hamsters, respectively. The order of the genes aligned on the long arm of human X chromosome was highly conserved in rat and the three rodent species except mouse. The present results suggest a possibility that the rat X chromosome retains the ancestral form of the rodent X chromosomes.
• Sequence polymorphisms, allelic expression status and chromosome locations of the chicken IGF2 and MPR1 genes

  T. Yokomine, A. Kuroiwa, K. Tanaka, M. Tsudzuki, Y. Matsuda, H. Sasaki

  Cytogenetic and Genome Research 93 1-2 109 - 113 2001年 

  By screening 26 chicken breeds and lines, DNA polymorphisms were identified in the <i>IGF2</i> and <i>MPR1</i> genes, of which mammalian homologues are parentally imprinted, and the <i>GAPD </i>gene, a housekeeping control. Using the polymorphisms as genetic markers, we found that all three genes are expressed biallelically in embryonic tissues. <i>IGF2</i> and <i>MPR1</i> were mapped on chicken chromosomes 5 and 3, respectively, by fluorescence in situ hybridization, demonstrating conserved linkage homology between mammals and birds.
• Molecular characterization of CRMP5, a novel member of the collapsin response mediator protein family

  M Fukada, Watakabe, I, J Yuasa-Kawada, H Kawachi, A Kuroiwa, Y Matsuda, M Noda

  JOURNAL OF BIOLOGICAL CHEMISTRY 275 48 37957 - 37965 2000年12月 [査読有り][通常論文]
   

  The CRMP (collapsin response mediator protein) family is thought to play key roles in growth cone guidance during neural development. The four members (CRMP1-4) identified to date have been demonstrated to form hetero-multimeric structures through mutual associations. In this study, we cloned a novel member of this family, which we call CRMP5, by the yeast two-hybrid method. This protein shares relatively low amino acid identity with the other CRMP members (49-50%) and also with dihydropyrimidinase (51%), whereas CRMP1-4 exhibit higher identity with each other (68-75%), suggesting that CRMP5 might be categorized into a third subfamily. The mouse CRMP5 gene was located at chromosome 5 B1, Northern blot and in situ hybridization analyses indicated that CRMP5 is expressed throughout the nervous system similarly to the other members (especially CRMP1 and CRMP4) with the expression peak in the first postnatal week. Association experiments using the yeast two-hybrid method and coimmunoprecipitation showed that CRMP5 interacts with dihydropyrimidinase and all the CRMPs including itself, except for CRMP1, although the expression profile almost overlaps with that of CRMP1 during development. These results suggest that CRMP complexes in the developing nervous system are classifiable into two populations that contain either CRMP1 or CRMP5. This indicates that different complexes may have distinct functions in shaping the neural networks.
• Cloning, genomic structure and chromosomal localization of the gene encoding mouse DNA helicase RecQ helicase protein-like 4

  T Ohhata, R Araki, R Fukumura, A Kuroiwa, Y Matsuda, K Tatsumi, M Abe

  GENE 261 2 251 - 258 2000年12月 [査読有り][通常論文]
   

  Five members of the RecQ helicase family, RECQL, WRN, BLM, RECQL4 and RECQL5 have been identified in humans. WRN and BLM have been demonstrated to be the responsible genes in Werner and Bloom syndromes, respectively. RECQL4 (RecQ helicase protein-like 4) was identified as a fourth member of the human RecQ helicase family bearing the helicase domain, and it was subsequently shown to be the responsible gene in Rothmund-Thomson syndrome. Here, we isolated mouse RECQL4 and determined the DNA sequence of full-length cDNA as well as the genome organization and chromosome locus. The mouse RECQL4 consists of 3651 base pairs coding 1216 amino acid residues and shares 63.4% of identical and 85.8% of homologous amino acid sequences with human RECQL4. The RECQL4 gene was localized to mouse chromosome 15D3 distal-E1 and rat chromosome 7q34 proximal. They were mapped in the region where the conserved linkage homology has been identified between the two species. Twenty-two exons dispersed over 7 kilo base pairs and all of the acceptor and donor sites for splicing of each exon conformed to the GT/AG rule. Our observations regarding mouse RECQL4 gene will contribute to functional studies on the RECQL4 products. (C) 2000 Elsevier Science B.V. All rights reserved.
• Assignment of human xylosylprotein β-1,4-galactosyltransferase gene (B4GALT7) to human chromosome 5q35.2→q35.3 by in situ hybridization

  A. Kuroiwa, Y. Matsuda, T. Okajima, K. Furukawa

  Cytogenetic and Genome Research 89 1-2 8 - 9 2000年06月26日
• Molecular analysis of avian circadian clock genes

  T Yoshimura, Y Suzuki, E Makino, T Suzuki, A Kuroiwa, Y Matsuda, T Namikawa, S Ebihara

  MOLECULAR BRAIN RESEARCH 78 1-2 207 - 215 2000年05月 [査読有り][通常論文]
   

  Unlike mammals, avian circadian rhythms are regulated by a multiple oscillatory system consisting of the retina, the pineal and the suprachiasmatic nucleus in the hypothalamus. To understand avian circadian system, we have cloned Clock and Period homologs (qClock, qPer2 and qPer3) and characterized these genes in Japanese quail. Overall, qCLOCK, qPER2 and qPER3 showed similar to 79%, similar to 46% and similar to 33% amino acid identity to mCLOCK, mPER2, mPER3, respectively. Clock was mapped to quail chromosome 4 and chicken chromosome 4q1.6-q2.1. Per2 and Per3 genes were both localized to microchromosomes. qClock mRNA was expressed throughout the day, while qPer2 and qPer3 showed robust circadian oscillation in the eye and the pineal gland. All three genes were expressed in various tissues. In addition, qPer2 mRNA was induced by light, but neither qClock nor qPer3 was induced. These results can explain the molecular basis for circadian entrainment in Japanese quail and also provide new avenues for molecular understanding of avian circadian clock and photoperiodism. (C) 2000 Elsevier Science B.V. All rights reserved.
• Identification and characterization of Elongin A2, a new member of the Elongin family of transcription elongation factors, specifically expressed in the testis

  T Aso, K Yamazaki, K Amimoto, A Kuroiwa, H Higashi, Y Matsuda, S Kitajima, M Hatakeyama

  JOURNAL OF BIOLOGICAL CHEMISTRY 275 9 6546 - 6552 2000年03月 [査読無し][通常論文]
   

  The Elongin complex stimulates the rate of transcription elongation by RNA polymerase II by suppressing the transient pausing of the polymerase at many sites along the DNA template. Elongin is composed of a transcriptionally active A subunit and two small regulatory B and C subunits, the latter of which bind stably to each other to form a binary complex that interacts with Elongin A and strongly induces its transcriptional activity. To further understand the roles of Elongin in transcriptional regulation, Re attempted to identify Elongin-related proteins. sere, we report on the cloning, expression, and characterization of human Elongin A2, a novel transcription elongation factor that exhibited 47% identity and 61% similarity to Elongin A. Biochemical studies have shown that Elongin A2 stimulates the rate of transcription elongation by RNA polymerase II and is capable of forming a stable complex with Elongin BC, However, in contrast to Elongin A, its transcriptional activity is not activated by Elongin BC. Northern blot analysis revealed that Elongin A2 mRNA was specifically expressed in the testis, suggesting that Elongin A2 may regulate the transcription of testis-specific genes.
• Molecular cloning, expression, and chromosomal mapping of human chondroitin 4-sulfotransferase, whose expression pattern in human tissues is different from that of chondroitin 6-sulfotransferase

  Tetsuya Okuda, Satoka Mita, Shinobu Yamauchi, Taeko Matsubara, Fumiko Yagi, Daiki Yamamori, Masakazu Fukuta, Asato Kuroiwa, Yoichi Matsuda, Osami Habuchi

  Journal of Biochemistry 128 5 763 - 770 2000年 [査読有り][通常論文]
   

  Chondroitin 4-sulfotransferase (C4ST) catalyzes the transfer of sulfate from 3'-phosphoadenosine 5'-phosphosulfate to position 4 of the N-acetylgalactosamine residues of chondroitin. We previously reported the cloning of C4ST cDNA from mouse brain. We here report the cloning and expression of human C4ST cDNA. The cDNA was isolated from a human fetal brain cDNA library by hybridization with a DNA probe prepared from rat poly(A)+ RNA used for the cloning of mouse C4ST cDNA. The cDNA comprises a single open reading frame that predicts a Type II transmembrane protein composed of 352 amino acids. The protein has an amino acid sequence homology of 96% with mouse C4ST. When the cDNA was introduced into a eukaryotic expression vector and transfected in COS-7 cells, the sulfotransferase activity that transfers sulfate to both chondroitin and desulfated dermatan sulfate was overexpressed. Northern blot analysis indicated that human C4ST mRNAs (6.0 and 1.9 kb) are expressed ubiquitously in various adult human tissues. Dot blot analysis has shown that human C4ST is strongly expressed in colorectal adenocarcinoma and peripheral blood leukocytes, whereas strong expression of human chondroitin 6-sulfotransferase (C6ST) is observed in aorta and testis. These observations suggest that the expression of C4ST and C6ST may be controlled differently in human tissues. The C4ST gene was localized to chromosome 12q23.2-q23.3 by fluorescence in situ hybridization.
• Assignment of human xylosylprotein β-1,4-galactosyltransferase gene(B4GALT7)to human chromosome 5q35.2→q35.3 by in situ hybridization

  Kuroiwa A, Matsuda Y, Okajima T, Furukawa K

  Cytogenetics Cell Genetics 89 1-2 8 - 9 2000年 [査読有り][通常論文]
  
• Mouse ULK2, a novel member of the UNC-51-like protein kinases: unique features of functional domains.

  Yan J, Kuroyanagi H, Tomemori T, Okazaki N, Asato K, Matsuda Y, Suzuki Y, Ohshima Y, Mitani S, Masuho Y, Shirasawa T, Muramatsu M

  Oncogene 18 43 5850 - 5859 1999年10月 [査読有り][通常論文]
  
• Cytogenetic mapping of 31 functional genes on chicken chromosomes by direct R-banding FISH

  T Suzuki, T Kurosaki, K Shimada, N Kansaku, U Kuhnlein, D Zadworny, K Agata, A Hashimoto, M Koide, M Koike, M Takata, A Kuroiwa, S Minai, T Namikawa, Y Matsuda

  CYTOGENETICS AND CELL GENETICS 87 1-2 32 - 40 1999年 [査読有り][通常論文]
   

  Using direct R-banding fluorescence in situ hybridization, we determined the location of 31 functional genes on chicken chromosomes. Replication R-banded chromosomes were obtained by synchronizing splenocyte cultures with excessive thymidine, followed by BrdU treatment. Thirty-one functional genes were directly localized to banded chicken chromosomes using genomic DNA and cDNA fragments as probes. The possibility of conserved linkage homology between chicken and human chromosomes was demonstrated for seven chicken chromosome regions (1p, 1q, 2q, 4p, 4q, and 5q). Copyright (C) 1999 S. Karger AG, Basel.
• 25.Green Fluorescent Protein (GFP) を用いた雌雄胚の判別

  中西 友子, 松田 洋一, 黒岩 麻里, 伊川 正人, 山田 秀一, 三輪 岳志, 岡部 勝

  日本疾患モデル学会記録 15 39 - 39 公益社団法人 日本実験動物学会 1999年
• Comparative FISH mapping of mouse and rat homologues of twenty-five human X-linked genes

  A. Kuroiwa,, T. Watanabe,, H. Hishigaki,, E. Takahashi,, T. Namikawa, Y. Matsuda

  Cytogenetic and Genome Research 81 3-4 208 - 212 1998年09月02日 

  We constructed a comparative cytogenetic map of 25 functional genes in mouse and rat X chromosomes by direct R-banding fluorescence in situ hybridization. Nineteen and 22 out of the 25 genes, which have been mapped on the human X chromosome, were newly localized to mouse and rat X chromosomes, respectively. Twenty-two additional genes were integrated in the rat-mouse-human comparative map of the X chromosome in this study. Comparison of the gene order indicated the presence of four chromosome segments with conserved linkage homology between mouse and rat X chromosomes, suggesting that a minimum of four chromosomal inversion events occurred between mouse and rat X chromosomes during the evolution of the two species. Four chromosome segments with conserved linkage homology were found between human and rat X chromosomes.
• Identification of mouse ULK1, a novel protein kinase structurally related to C-elegans UNC-51

  J Yan, H Kuroyanagi, A Kuroiwa, Y Matsuda, H Tokumitsu, T Tomoda, T Shirasawa, M Muramatsu

  BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 246 1 222 - 227 1998年05月 [査読有り][通常論文]
   

  A novel protein kinase related to the C. elegans serine/threonine kinase UNC-51 was cloned from mouse. The UNC-51-Like Kinase (ULK)1 is encoded by a cDNA of 1051 amino acids with calculated MW of 113 kDa. Comparison of the ULK1 and UNC-51 shows the highest conservation in the amino-terminal kinase domain, which is followed by a proline/serine-rich (PS) domain and a conserved carboxyl-terminal (C) domain. ULK1 mRNA is expressed in various tissues, and is mapped to mouse chromosome 5F and rat chromosome 12q16.3, by fluorescent in situ hybridization. PIA-tagged ULK1 is expressed as a protein of similar to 150 kDa in COS7 cells and is auto-phosphorylated in vitro in its PS domain. We propose that ULK1, UNC-51 and a yeast protein kinase Apg1p comprise a novel subfamily of protein kinase, which is structurally conserved among eukaryotes. (C) 1998 Academic Press.
• Association of tyrosine phosphatases SHP-1 and SHP-2, inositol 5-phosphatase SHIP with gp49B1, and chromosomal assignment of the gene

  A Kuroiwa, Y Yamashita, M Inui, T Yuasa, M Ono, A Nagabukuro, Y Matsuda, T Takai

  JOURNAL OF BIOLOGICAL CHEMISTRY 273 2 1070 - 1074 1998年01月 [査読無し][通常論文]
   

  We have analyzed the molecules participating in the inhibitory function of gp49B1, a murine type I transmembrane glycoprotein expressed on mast cells and natural killer cells, as well as the chromosomal location of its gene. As assessed by SDS-polyacrylamide gel electrophoresis and immunoblot analysis, tyrosine-phosphorylated, but not nonphosphorylated, synthetic peptides matching each of the two immunoreceptor tyrosine-based inhibitory motif (ITIM)-like sequences found in the cytoplasmic portion of gp49B1 associated with the similar to 65-kDa tyrosine phosphatase SHP-1 and similar to 70-kDa SHP-2 derived from RBL-2H3 cells, In addition, the phosphotyrosyl peptide matching the second ITIM-like sequence also bound the similar to 145-kDa inositol polyphosphate 5-phosphatase SHIP. Thus, it has been strongly suggested that the inhibitory nature of gp49B involves the recruitment of SHP-1, SHP-2, and SHIP for the delivery of inhibitory signal to the cell interior upon phosphorylation of tyrosine residues in their ITIMs. The gp49B gene has been found to be in the juxtaposition of its cognate gene, gp49A. The gene pair was shown to locate in the B4 band of mouse chromosome 10. In this region, no conserved linkage homology to human chromosome 19, where the genes for killer cell inhibitory receptors are found, has been identified.
• Comparative FISH mapping of mouse and rat homologues of twenty-five human X-linked genes

  A Kuroiwa, T Watanabe, H Hishigaki, E Takahashi, T Namikawa, Y Matsuda

  CYTOGENETICS AND CELL GENETICS 81 3-4 208 - 212 1998年 [査読有り][通常論文]
   

  We constructed a comparative cytogenetic map of 25 functional genes in mouse and rat X chromosomes by direct R-banding fluorescence in situ hybridization. Nineteen and 22 out of the 25 genes, which have been mapped on the human X chromosome, were newly localized to mouse and rat X chromosomes, respectively. Twenty-two additional genes were integrated in the rat-mouse-human comparative map of the X chromosome in this study. Comparison of the gene order indicated the presence of four chromosome segments with conserved linkage homology between mouse and rat X chromosomes, suggesting that a minimum of four chromosomal inversion events occurred between mouse and rat X chromosomes during the evolution of the two species. Four chromosome segments with conserved linkage homology were found between human and rat X chromosomes.
• FISH 法を用いた染色体マッピング

  松田 洋一, 黒岩 麻里

  アニテックス 10 20 - 24 1998年 [査読有り][招待有り]
  
• Chromosomal mapping of the gene encoding serotonin N-acetyltransferase to rat chromosome 10q32.3 and mouse Chromosome 11E2

  T Yoshimura, A Nagabukuro, Y Matsuda, T Suzuki, A Kuroiwa, M Iigo, T Namikawa, S Ebihara

  CYTOGENETICS AND CELL GENETICS 79 3-4 172 - 175 1997年 [査読有り][通常論文]
   

  Pineal melatonin is produced during the night. Its nocturnal increase regulates circadian rhythms and the photoperiodic reproductive response. Serotonin is acetylated to N-acetylserotonin by serotonin N-acetyltransferase (SNAT) and then methylated to form melatonin by hydroxyindole-O-methyltransferase (HIOMT). The rhythmicity of melatonin synthesis is regulated by the rhythmic activity of SNAT. Most laboratory mice do not have melatonin because of a genetic defect in the activity of SNAT and/or HIOMT. In a previous study using a recombinant inbred strain, we have found that the locus controlling pineal SNAT activity (Nat4) is located on mouse Chromosome 11. Recently, SNAT has been cloned in the rat. In the present study, the gene encoding SNAT was localized, using a rat cDNA fragment, on rat and mouse chromosomes by direct R-banding fluorescence in situ hybridization (FISH). In addition, using molecular linkage analysis with interspecific backcross mice, a gene encoding SNAT was mapped on a mouse chromosome. The gene encoding SNAT was localized to rat chromosome 10q32.3 and mouse Chromosome 11E2 by FISH. The molecular linkage analysis demonstrated that the gene encoding SNAT maps 1.5 cM distal to D11Mit11. The data suggest that Natl encodes SNAT. These chromosomal locations are in a region of conserved linkage homology between the two species.
• Chromosomal mapping of the gene encoding serotonin N-acetyltransferase to rat chromosome 10q32.3 and mouse Chromosome 11E2

  Takashi Yoshimura, A. Nagabukuro, Y. Matsuda, T. Suzuki, A. Kuroiwa, M. Ligo, T. Namikawa, S. Ebihara

  Cytogenetic and Genome Research 79 3-4 172 - 175 1997年01月01日 [査読有り][通常論文]
   

  Pineal melatonin is produced during the night. Its nocturnal increase regulates circadian rhythms and the photo-periodic reproductive response. Serotonin is acetylated to N-acetylserotonin by serotonin N-acetyltransferase (SNAT) and then methylated to form melatonin by hydroxyindole-O-methyltransferase (HIOMT). The rhythmicity of melatonm synthesis is regulated by the rhythmic activity of SNAT. Most laboratory mice do not have melatonin because of a genetic defect in the activity of SNAT and/or HIOMT. In a previous study using a recombinant inbred strain, we have found that the locus controlling pineal SNAT activity (Nat4) is located on mouse Chromosome 11. Recently, SNAT has been cloned in the rat. In the present study, the gene encoding SNAT was localized, using a rat cDNA fragment, on rat and mouse chromosomes by direct R-banding fluorescence in situ hybridization (FISH). In addition, using molecular linkage analysis with interspecific back-cross mice, a gene encoding SNAT was mapped on a mouse chromosome. The gene encoding SNAT was localized to rat chromosome 10q32.3 and mouse Chromosome 11E2 by FISH. The molecular linkage analysis demonstrated that the gene encoding SNAT maps 1.5 cM distal to DUMit11. The data suggest that Nat4 encodes SNAT. These chromosomal locations are in a region of conserved linkage homology between the two species. © 1997 S. Karger AG, Basel.

書籍

• Reproductive and Developmental Strategies; the Continuity of Life

  KUROIWA Asato (担当:共著範囲:Sex determination and differentiation in birds)
  Springer Japan 2018年
• Avian Reproduction: From Behavior to Molecules

  KUROIWA Asato (担当:共著範囲:Sex-determining mechanism in avian)
  Springer Japan 2017年06月
• ホルモンから見た生命現象と進化シリーズ 第3巻 成長・成熟・性決定 — 継 —, 日本比較内分泌学会編

  黒岩 麻里 (担当:共著範囲:鳥類の性決定と性成熟)
  裳華房 2016年05月
• (080)男の弱まり (ポプラ新書)

  黒岩 麻里 (担当:単著)
  ポプラ社 2016年01月 (ISBN: 459114738X) 250

  
• 消えゆくY染色体と男たちの運命ーオトコの生物学

  黒岩 麻里 (担当:単著)
  学研メディカル秀潤社 2014年03月 (ISBN: 4780908922) 223

  
• Sex Chromosomes: New Research" (eds: Mario D'Aquino & Vincente Stallone)

  KUROIWA Asato (担当:共著範囲:The fate of the Y chromosome)
  Nova Publisher's Inc 2012年
• 生物多様性の基礎知識 (草刈秀紀 編著)

  黒岩 麻里 (担当:共著)
  日刊工業新聞社 2010年08月
• The Wild Mammals of Japan. (ed. Ohdachi SD, Ishibashi Y, Iwasa MA, Saitoh T)

  KUROIWA Asato (担当:共著範囲:Unique and interesting sex chromosome evolution in Tokudaia)
  Mammalogical Society of Japan 2009年
• FISH法を用いた染色体マッピング

  別冊実験医学、non-RI実験の最新プロトコール、羊土社 1999年

講演・口頭発表等

• Y染色体の役割と運命−Yをもたない哺乳類の性決定  [招待講演]

  黒岩 麻里

  染色体学会第72回年会 市民公開講座「みんなに知ってもらいたい、最新の染色体研究」 2021年09月 公開講演，セミナー，チュートリアル，講習，講義等
• Y染色体消失と性決定の多様性  [招待講演]

  黒岩 麻里

  日本進化学会第23回東京大会 シンポジウム「動物の性決定システムの進化」 2021年08月 シンポジウム・ワークショップパネル（指名）
• Unique genome evolution of the Y-absent mammal in genus Tokudaia  [通常講演]

  Asato Kuroiwa (presentar/organizer)

  “Biological fate determination by multidimensional genome changes”, The 43rd Annual Meeting of the Molecular Biology Society of Japan. 2020年12月 シンポジウム・ワークショップパネル（指名）
• Y染色体とSRY遺伝子をもたない哺乳類種の性決定メカニズム  [招待講演]

  黒岩麻里

  第92回日本遺伝学会年会，ワークショップ「マウス遺伝学からみるクロマチン制御機構」 2020年09月 シンポジウム・ワークショップパネル（指名）
• トゲネズミ属におけるSRY遺伝子に依存しない性決定の分子メカニズム  [招待講演]

  黒岩麻里, 奥野未来, 伊藤武彦, 寺尾美穂, 小川湧也, 高田修治, 水島秀成

  2019年12月 シンポジウム・ワークショップパネル（指名）
• Y染色体の役割と運命−Yをもたない哺乳類の性決定  [招待講演]

  黒岩 麻里

  第112回日本繁殖生物学会市民公開講座「性におけるオスとメスの役割に関する新展開」 2019年09月 公開講演，セミナー，チュートリアル，講習，講義等
• Y染色体をもたない哺乳類種の性染色体と性決定機構の進化  [招待講演]

  黒岩 麻里

  遺伝研研究会「有性生殖にかかわる染色体・クロマチン・核動態に関する研究会」 2019年06月 口頭発表（招待・特別）
• トゲネズミ属におけるSRY遺伝子の機能と進化  [通常講演]

  黒岩 麻里

  第41回日本分子生物学会年会 2018年11月 シンポジウム・ワークショップパネル（指名）
• Unique sex chromosome and sex-determining mechanism in Japanese native mammals, genus Tokudaia.  [招待講演]

  KUROIWA Asato

  Genetic Society of Australia 2018 and 6th Asia-Pacific Chromosome Colloquium (GSA2018_APCC6) 2018年07月 口頭発表（基調）
• Sex chromosome evolution and sex-determining mechanism in SRY-absent XO/XO mammals, genus Tokudaia.  [招待講演]

  KUROIWA Asato

  8th Internationa symposium on the biology of vertebrate sex determination 2018年04月 口頭発表（招待・特別）
• XとYのミステリー 性決定の不思議  [招待講演]

  黒岩 麻里

  池田町シニアカレッジ遊ゆう大学 2018年02月 公開講演，セミナー，チュートリアル，講習，講義等
• SRY遺伝子をもたない哺乳類種の新しい性決定メカニズム  [招待講演]

  黒岩 麻里

  生命科学系学会合同年次大会 (ConBio2017) 2017年12月 口頭発表（招待・特別）
• ワークショップ“またまたやってきたオモロイ生き物の分子生物学”  [通常講演]

  黒岩 麻里, 三浦 恭子, オーガナイザー

  生命科学系学会合同年次大会 (ConBio2017) 2017年12月 シンポジウム・ワークショップパネル（公募）
• XとYのミステリー 性が決まる仕組みの生物学  [招待講演]

  黒岩 麻里

  北海道札幌啓成高等学校“SSH特別科学講演会” 2017年09月 公開講演，セミナー，チュートリアル，講習，講義等
• トゲネズミの性決定機構—これまでに明らかになった現象  [招待講演]

  黒岩 麻里

  日本哺乳類学会2017年度大会 2017年09月 口頭発表（招待・特別）
• Y染色体を失った哺乳類の性決定メカニズム  [招待講演]

  黒岩 麻里

  国立成育医療研究センター特別セミナー 2017年07月 口頭発表（招待・特別）
• トゲネズミ属におけるSRY遺伝子の機能消失  [招待講演]

  黒岩 麻里

  第39回日本分子生物学会年会 2016年11月 口頭発表（招待・特別）
• XとYのはたらき—ヒトの性差のつくられ方  [招待講演]

  黒岩 麻里

  北海道女性協会主催 “えるのす連続講座” 2016年11月 公開講演，セミナー，チュートリアル，講習，講義等
• ニワトリの性決定に関わる新規遺伝子の発見  [招待講演]

  黒岩 麻里

  第159回日本獣医学会学術集会 2016年09月 口頭発表（招待・特別）
• 消えゆくY染色体の運命  [招待講演]

  黒岩 麻里

  第4回関西生殖医学集談会/第48回関西アンドロロジーカンファレンス 2016年03月 口頭発表（招待・特別）
• Y染色体をもたない哺乳類の性決定メカニズム  [招待講演]

  黒岩 麻里

  北大・産総研若手研究者研究交流会 2016年02月 口頭発表（招待・特別）
• 動物の染色体の観察と同定―核型から見える種の多様性  [招待講演]

  黒岩 麻里

  北海道立教育研究所附属理科教育センター主催 “理科特別演習講座” 2016年01月 公開講演，セミナー，チュートリアル，講習，講義等
• Yをすてた日本のネズミ―SRYをもたない哺乳類の性決定メカニズム  [招待講演]

  黒岩 麻里

  第38回日本分子生物学会 2015年12月 口頭発表（招待・特別）
• ニワトリの性決定に関わる新規遺伝子の発見  [招待講演]

  黒岩 麻里

  岩手大学全学共通教育「自然科目委員会」FD活動講演会 2015年11月 公開講演，セミナー，チュートリアル，講習，講義等
• 哺乳類Y染色体の消失過程の推定  [招待講演]

  黒岩 麻里

  日本遺伝学会第87回大会 2015年09月 口頭発表（招待・特別）
• Genomic properties of the Ryukyu spiny rats (genus Tokudaia) and evolutionary perspectives.  [招待講演]

  KUROIWA Asato

  Vth International Wildlife Management Congress (IWMC2015) 2015年07月 口頭発表（招待・特別）
• Y染色体をもたない哺乳類の性決定メカニズム  [招待講演]

  黒岩 麻里

  第50回北陸実験動物研究会 2015年07月 口頭発表（招待・特別）
• XとYのミステリー〜性決定の不思議〜  [招待講演]

  黒岩 麻里

  北海道生涯学習協会主催 “「北海道学」かでる講座（道民カレッジ連携講座）” 2015年06月 公開講演，セミナー，チュートリアル，講習，講義等
• Evolution of sex chromosomes and sex-determining mechanism in Y-absent mammals.  [招待講演]

  KUROIWA Asato

  International Symposium of Correlative Gene System Establishing Next-Generation Genetics 2015年05月 口頭発表（招待・特別）
• Evolution of the sex chromosomes in Y-absent mammals.  [招待講演]

  KUROIWA Asato

  Asian Chromosome Colloquium 2015 (ACC5) 2015年04月 口頭発表（招待・特別）
• ワークショップ「脊椎動物の性分化分子機構」  [通常講演]

  黒岩 麻里, 高田 修治, オーガナイザー

  第37回日本分子生物学会 2014年11月 シンポジウム・ワークショップパネル（公募）
• 男性はどこへ？Y染色体の運命  [招待講演]

  黒岩 麻里

  財団法人染色体学会主催，市民公開講座 “ゲノムと性—オスとメスを決めるからくり” 2014年10月 公開講演，セミナー，チュートリアル，講習，講義等
• ワークショップ 「新しい性染色体の獲得と進化」  [通常講演]

  黒岩 麻里, 寺井 洋平, オーガナイザー

  日本遺伝学会第86回大会 2014年09月 シンポジウム・ワークショップパネル（公募）
• 儚きY染色体と男たちの運命  [招待講演]

  黒岩 麻里

  河合塾主催，河合塾生物学セミナー 2014年08月 公開講演，セミナー，チュートリアル，講習，講義等
• Y染色体をもたないトゲネズミの性決定メカニズム  [招待講演]

  黒岩 麻里

  日本実験動物科学技術さっぽろ2014 2014年05月 口頭発表（招待・特別）
• シンポジウム「性染色体がうまれるとき」  [通常講演]

  黒岩 麻里, オーガナイザー

  FResHU F3 シンポジウム 2014年03月 シンポジウム・ワークショップパネル（公募）
• 性を決める遺伝子  [招待講演]

  黒岩 麻里

  文部科学省新学術領域「性差構築の分子基盤」主催，市民公開シンポジウム “性の不思議―女と男―” 2013年12月 公開講演，セミナー，チュートリアル，講習，講義等
• 生物の性が決まる仕組み  [招待講演]

  黒岩 麻里

  第107回環境・自然を考える会 2013年11月 公開講演，セミナー，チュートリアル，講習，講義等
• ニワトリの精巣分化に関わる新規遺伝子の解析  [招待講演]

  黒岩 麻里

  日本動物遺伝育種学会第14回大会 2013年10月 口頭発表（招待・特別）
• Sex chromosome evolution in Y-absent mammals.  [招待講演]

  KUROIWA Asato

  International symposium on “molecular and phenotype evolution” 2013年09月 口頭発表（招待・特別）
• ニワトリ性分化に関わる新規遺伝子の機能解明―過剰発現TGニワトリ胚の解析―  [招待講演]

  黒岩 麻里

  日本分子生物学会第35回年会 2012年12月 口頭発表（招待・特別）
• Y染色体をもたない哺乳類の進化研究  [招待講演]

  黒岩 麻里

  北海道牛受精卵移植研究会 第31回研究発表会 2012年08月 口頭発表（招待・特別）
• CHH is a new gene involved in the gonadal differentiation of chicken.  [招待講演]

  KUROIWA Asato

  10th International Symposium on Avian Endcrinology 2012年06月 口頭発表（招待・特別）
• Sex-determining mechanism of birds  [通常講演]

  KUROIWA Asato (o

  FResHU F3 Green Symposia Series #3 2012年05月 シンポジウム・ワークショップパネル（公募）
• 性決定と性比  [招待講演]

  黒岩 麻里

  応用倫理研究会 2011年12月 公開講演，セミナー，チュートリアル，講習，講義等
• 性を決める遺伝子  [招待講演]

  黒岩 麻里

  文部科学省新学術領域「性差構築の分子基盤」主催，市民公開シンポジウム “性の不思議―女と男―” 2011年09月 公開講演，セミナー，チュートリアル，講習，講義等
• 染色体から読み解く性の未来  [招待講演]

  黒岩 麻里

  男女共同参画企画事業交流会 2011年02月 公開講演，セミナー，チュートリアル，講習，講義等
• Y染色体の進化—消失か？存続か？  [招待講演]

  黒岩 麻里

  分子生物学会・生化学会・生物物理学会合同シンポジウム 2010年11月 口頭発表（招待・特別）
• 性を決める遺伝子と性染色体のしくみ  [招待講演]

  黒岩 麻里

  財団法人染色体学会主催，市民公開講座 “知っておきたい身近な‘遺伝子と染色体’のはなし” 2010年11月 公開講演，セミナー，チュートリアル，講習，講義等
• 天然記念物トゲネズミの保全活動と遺伝学  [招待講演]

  黒岩 麻里

  日本遺伝学会主催，市民公開講座 “遺伝学は語る〜未来へのメッセージ” 2010年09月 公開講演，セミナー，チュートリアル，講習，講義等
• ワークショップ「Y染色体の進化」  [通常講演]

  黒岩 麻里, 黒木 陽子

  2010年09月 シンポジウム・ワークショップパネル（公募）
• 分子細胞遺伝学：生物としての男  [招待講演]

  黒岩 麻里

  さっぽろ自由学校「遊」公開講座 2008年10月 公開講演，セミナー，チュートリアル，講習，講義等
• トゲネズミの保全活動と染色体研究  [招待講演]

  黒岩 麻里

  日本動物学会北海道支部主催，公開講演会 “動物学への招待” 2008年08月 公開講演，セミナー，チュートリアル，講習，講義等
• アマミトゲネズミにおけるY染色体消失過程の推定―Y連鎖遺伝子の運命―  [招待講演]

  黒岩 麻里

  第11回遺伝学談話会 2008年05月 口頭発表（招待・特別）
• トゲネズミたちの不思議―染色体のはなし―  [招待講演]

  黒岩 麻里

  環境省やんばる野生生物保護センター主催，公開講演会 2008年05月 公開講演，セミナー，チュートリアル，講習，講義等
• Sex chromosome evolution in the X0 mammal, the Amami spiny rat (Tokudaia osimensis).  [招待講演]

  KUROIWA Asato

  The 52nd NIBB Conferences 2006年01月 口頭発表（招待・特別）
• 哺乳類における性染色体の分化と性決定機構の進化  [招待講演]

  黒岩 麻里

  財団法人染色体学会主催，公開シンポジウム “性の分化について考える〜性染色体研究の現状と展望” 2005年10月 公開講演，セミナー，チュートリアル，講習，講義等
• 鳥類のエピジェネシス―鳥類の性染色体に遺伝子量補正機構は存在するのか―  [招待講演]

  黒岩 麻里, 松田 洋一

  日本畜産学会第100回大会 2002年03月 口頭発表（招待・特別）
• 齧歯類及び食虫類の染色体構造進化―特にＸ染色体の構造変化を中心として―  [招待講演]

  黒岩 麻里, 松田 洋一

  日本遺伝学会第70回大会 1998年09月 口頭発表（招待・特別）

その他活動・業績

• トゲネズミ研究の最近（4）〜保全と生命科学研究を繋ぐ〜

  城ヶ原貴通, 越本知大, 安田雅俊, 小高信彦, 黒岩麻里 哺乳類科学 58 (1) 103 -104 2018年 [査読無し][通常論文]
  
• トゲネズミ研究の最近3〜琉球諸島哺乳類保全の次世代を担う者達〜

  城ヶ原貴通, 山田文雄, 越本知大, 黒岩麻里, 木戸文香, 中家雅隆, 望月春佳, 村田知慧, 三谷 匡 哺乳類科学 53 (1) 170 -173 2013年 [査読無し][通常論文]
  
• トゲネズミ研究の最近（2）

  城ヶ原貴通, 山田文雄, 村田智慧, 黒岩麻里, 越本知大, 三谷匡 哺乳類科学 51 (1) 154 -158 2011年 [査読無し][通常論文]
  
• オキナワトゲネズミ再発見と, トゲネズミ研究の最近

  山田 文雄, 鈴木 仁, 黒岩 麻里, 村田 知慧 哺乳類科学 49 (1) 133 -135 2009年06月30日 [査読無し][通常論文]
  
• Chromosome evolution of the cichlid fishes from Lake Victoria

  K. Yoshida, A. Kuroiwa, Y. Terai, S. Mizoiri, M. Aibara, N. Kobayashi, Y. Matsuda, N. Okada INTEGRATIVE AND COMPARATIVE BIOLOGY 49 E329 -E329 2009年02月 [査読無し][通常論文]
  
• O-15. Increase in the copy number of the M33 gene of Ryukyu spiny rat(Abstracts of the oral and poster presentations) :

  HANDA Sanae, YAMADA Fumio, HASHIMOTO Takuma, ABE Shintaro, SHINOHARA Akio, KOSHIMOTO Chihiro, KATOH-FUKUI Yuko, MOROHASHI Ken-Ichirou, MATSUDA Yoichi, KUROIWA Asato Chromosome science 9 (4) 119 -119 2006年
• O-14. Chromosomal localization of mammalian Y-linked genes in X0/X0 mammal, Ryukyu spiny rat(Abstracts of the oral and poster presentations) :

  ISHIGUCHI Yasuko, YAMADA Fumio, HASHIMOTO Takuma, ABE Shintaro, MATSUDA Yoichi, KUROIWA Asato Chromosome science 9 (4) 118 -118 2006年
• O-04. Development of chromosome markers in a cichlid species, Paralabidochromis chilotes, from Lake Victoria in Africa(Abstracts of the oral and poster presentations) :

  KUROIWA Asato, WATANABE Masakatsu, OKADA Norihiro, MATSUDA Yoichi Chromosome science 9 (4) 113 -113 2006年
• Sex chromosome differentiation and sex determination in mammals(Symposium open to the public : "Thinking about 'Sex Differentiation' : the Present Status and Prospect of Chromosome Research", Abstract of the 56th Annual Meeting of The Society of Chromosome Research) :

  Kuroiwa Asato Chromosome science 9 (1) 8 -8 2006年
• O-24. Detection of chromosomal differences between female and male Amami spiny rat, XO(Abstracts of the oral and poster presentations, Abstract of the 56th Annual Meeting of The Society of Chromosome Research) :

  KOBAYASHI Tsuyoshi, YAMADA Fumio, HASHIMOTO Takuma, SATO Kaori, ISHII Nobuo, ABE Shintaro, SHINOHARA Akio, KOSHIMOTO Chihiro, MATSUDA Yoichi, KUROIWA Asato Chromosome science 9 (1) 21 -21 2006年
• O-07. Search of sex determination gene in Amami spiny rat, XO(Abstracts of the oral and poster presentations, Abstract of the 56th Annual Meeting of The Society of Chromosome Research) :

  HANDA Sanae, YAMADA Fumio, HASHIMOTO Takuma, SATO Kaori, ISHII Nobuo, ABE Shintaro, SHINOHARA Akio, KOSHIMOTO Chihiro, KATOH(FUKUI) Yuko, MOROHASHI Ken-Ichirou, MATSUDA Yoichi, KUROIWA Asato Chromosome science 9 (1) 13 -13 2006年
• O-13. Centromere repositioning in X chromosome of X0/X0 mammals, Ryukyu spiny rat(Abstracts of the oral and poster presentations)

  Tsuyoshi KOBAYASHI, Fumio YAMADA, Takuma HASHIMOTO, Shintaro ABE, Akio SHINOHARA, Chihiro KOSHIMOTO, Yoichi MATSUDA, Asato KUROIWA, Grad. Sch, Sci. Hokkaido Univ, Kansai Res. Cen. FFPRI, JWRC, Amami Wildlife, Conser. Cen, Min. Envi, Frontier Sci, Res. Cen, Miyazaki Univ, Frontier Sci, Res. Cen, Miyazaki Univ, Grad. Sch, Sci. Hokkaido, Univ.:CRIS Hokkaido Univ, Grad. Sch, Sci. Hokkaido, Univ.:CRIS Hokkaido Uni Chromosome science 9 (4) 2006年01月01日 [査読無し][通常論文]
  
• イモリにおける核内遺伝子配置を可視化する方法の開発

  牧信安, 黒岩麻里, 梅原千鶴子, 松田洋一, 阿形清和 日本発生生物学会大会発表要旨集 36th 2003年
• Studies on mammalian karyotypic evolution by comparative FISH analyses(Symposium : Current development and perspective on chromosome banding techniques)(The 53rd Annual Meeting of the Society of Chromosome Research) :

  Matsubara Kazumi, Kuroiwa Asato Chromosome science 6 (4) 107 -107 2002年
• Identification of two different isoforms of mouse ADAM 1 (fertilin α)

  NISHIMURA Hitoshi, KIM Ekyune, FUJIMORI Tetsuji, KASHIWABARA Shin-ichi, KUROIWA Asato, MATSUDA Yoichi, BABA Tadashi Keio journal of medicine 50 83 -83 2001年10月01日
• 自由30 FISH法を用いた遺伝子オーダリングに基づく霊長類X染色体の構造変化に関する研究(VI 共同利用研究 2.研究成果)

  黒岩 麻里, 松田 洋一 霊長類研究所年報 31 154 -154 2001年10月01日
• Zoo‐FISH法を用いたApodemus属の核型進化の解析

  松原和純, 梅原千鶴子, 黒岩麻里, 土屋公幸, 松田洋一 日本遺伝学会大会プログラム・予稿集 73rd 131 2001年08月22日 [査読無し][通常論文]
  
• 自由27 FISH法を用いた遺伝子オーダリングに基づく霊長類X染色体の構造変化に関する研究(V 共同利用研究 2.研究成果)

  黒岩 麻里, 松田 洋一 霊長類研究所年報 30 126 -126 2000年10月01日

受賞

• 2023年10月 国立大学法人北海道大学 桂田芳枝賞
  
• 2013年04月 文部科学省 平成25年度文部科学大臣表彰若手科学者賞
   Y染色体をもたない哺乳類種の性染色体進化の研究 

  受賞者: 黒岩 麻里
• 2011年11月 財団法人染色体学会 2011年度(第62回)染色体学会賞
   哺乳類および鳥類における性染色体と性決定機構の進化研究 

  受賞者: 黒岩 麻里

共同研究・競争的資金等の研究課題

• シス因子による哺乳類の新しい性決定メカニズムの解明

  日本学術振興会：科学研究費助成事業

  研究期間 : 2022年04月 -2026年03月 

  代表者 : 黒岩 麻里
• 化学物質の鳥類卵内投与による性分化異常評価手法の開発とテストガイ ドライン化に向けた提案

  環境研究総合推進費

  研究期間 : 2022年04月 -2025年03月
• 有胎盤哺乳類におけるSRY遺伝子に依存しない新しい性決定メカニズムの解明

  日本学術振興会：科学研究費助成事業 基盤研究(B)

  研究期間 : 2019年04月 -2022年03月 

  代表者 : 黒岩 麻里, 高田 修治, 伊藤 武彦
• 鳥類の性決定にはたらくnon-coding RNAの解析

  日本学術振興会：科学研究費補助金 挑戦的研究（萌芽）

  研究期間 : 2018年06月 -2020年03月 

  代表者 : 黒岩 麻里
• XO型アマミトゲネズミにおける元Y遺伝子の解析

  成茂動物科学振興基金：成茂動物科学振興基金研究助成

  研究期間 : 2018年10月 

  代表者 : 黒岩 麻里
• 平胸類エミューを用いた鳥類の性決定遺伝子の同定

  日本学術振興会：新学術領域研究生命科学系3分野支援活動 ゲノム支援

  研究期間 : 2017年06月 -2018年03月 

  代表者 : 黒岩 麻里
• 平胸類エミューを用いた鳥類の性決定遺伝子の同定

  日本学術振興会：科学研究費補助金 基盤研究（B）

  研究期間 : 2015年04月 -2018年03月 

  代表者 : 黒岩 麻里
• Chicken and ChIPs; genetic conrol of avian gonadal development

  4. Australian Government, Australian Research Council：Discovery Projects Proposal for Funding Commencing, 2016〜2018

  研究期間 : 2016年 -2018年 

  代表者 : Craig Smith

   

  Partner Investigator: Asato Kuroiwa Chief Investigator: Craig Smith (Monash University)
• 平胸類エミューを用いた鳥類の性決定遺伝子の同定

  日本学術振興会：新学術領域研究生命科学系3分野支援活動 ゲノム支援

  研究期間 : 2016年04月 -2017年03月 

  代表者 : 黒岩 麻里
• 性分化疾患の解明に向けたSOX9遺伝子遠位エンハンサーの解析

  公益財団法人 寿原記念財団：寿原記念財団研究助成

  研究期間 : 2017年 

  代表者 : 黒岩 麻里
• SRYをもたない哺乳類における新しい性決定遺伝子の同定

  日本学術振興会：科学研究費補助金 新学術領域研究(研究領域提案型)

  研究期間 : 2014年04月 -2016年03月 

  代表者 : 黒岩 麻里
• SRYをもたない哺乳類における新しい性決定遺伝子の同定

  日本学術振興会：新学術領域研究生命科学系3分野支援活動 ゲノム支援

  研究期間 : 2014年06月 -2015年03月 

  代表者 : 黒岩 麻里
• 鳥類特異的な生殖腺性差構築に関わる新規遺伝子の解析

  日本学術振興会：科学研究費補助金 新学術領域研究（研究領域提案型）

  研究期間 : 2011年04月 -2013年03月 

  代表者 : 黒岩 麻里
• Y染色体退化と雄性機能維持メカニズムの解明

  内藤記念財団：内藤記念女性研究者研究助成金

  研究期間 : 2011年 -2013年 

  代表者 : 黒岩 麻里
• ニワトリの性決定遺伝子の同定

  日本学術振興会：科学研究費補助金 挑戦的萌芽研究

  研究期間 : 2010年04月 -2012年03月 

  代表者 : 黒岩 麻里
• 性決定機構が未解明な動物種における性染色体の構造と性決定関連遺伝子群の解析

  日本学術振興会：科学研究費補助金 特定領域研究

  研究期間 : 2004年04月 -2009年03月 

  代表者 : 松田 洋一

   

  研究分担者 黒岩 麻里
• 集団遺伝学を取り入れた種形成機構の解析

  日本学術振興会：科学研究費補助金 特定領域研究

  研究期間 : 2006年04月 -2008年03月 

  代表者 : 舘田 英典
• XO型アマミトゲネズミにおけるX染色体不活性化機構の研究

  日本学術振興会：科学研究費補助金 若手研究(B)

  研究期間 : 2006年04月 -2008年03月 

  代表者 : 黒岩 麻里
• トゲネズミ三種の比較解析によるY染色体消失過程の推定

  住友財団：住友財団基礎科学研究助成

  研究期間 : 2008年 

  代表者 : 黒岩 麻里
• 絶滅危惧種オキナワトゲネズミの捕獲調査と研究材料の確保

  北海道大学：北海道大学若手研究者自立支援

  研究期間 : 2008年 

  代表者 : 黒岩 麻里
• SRY遺伝子に依存しない新たな性決定メカニズムの解明

  内藤記念財団：第40回内藤記念科学奨励金（研究助成）

  研究期間 : 2008年 

  代表者 : 黒岩 麻里
• XO型トゲネズミにおける性染色体進化の研究

  稲盛財団：稲盛財団研究助成

  研究期間 : 2006年 

  代表者 : 黒岩 麻里
• ニワトリゲノムにおける遺伝子量補正機構関連遺伝子の探索

  ノーステック財団：ノーステック財団基盤的研究開発育成事業 (若手研究補助金)

  研究期間 : 2004年 

  代表者 : 黒岩 麻里
• ニワトリの遺伝子量補正機構に関与する新規遺伝子の探索

  秋山記念財団：秋山記念生命科学研究助成金

  研究期間 : 2004年 

  代表者 : 黒岩 麻里
• 鳥類の性染色体における遺伝子量補正機構の解析

  日本学術振興会：科学研究費補助金 特別研究員奨励費

  研究期間 : 2003年04月 

  代表者 : 黒岩 麻里
• 鳥類における性染色体不活性化機構の解析

  日本学術振興会：科学研究費補助金 特別研究員奨励費

  研究期間 : 2001年04月 -2003年03月 

  代表者 : 黒岩 麻里
• A study on gene dosage compensation of Z chromosome in chicken.

教育活動情報

主要な担当授業

大学運営

学内役職歴

• 2017年10月26日 - 2019年3月31日 経営戦略室室員
• 2019年4月1日 - 2020年9月30日 経営戦略室室員
• 2020年10月12日 - 2021年3月31日 経営戦略室室員
• 2022年4月1日 - 2023年1月9日 広報・社会連携室室員
• 2023年1月10日 - 2024年3月31日 広報・社会連携室室員
• 2020年10月12日 - 2022年3月31日 広報室室員
• 2017年4月1日 - 2019年3月31日 総長補佐
• 2019年4月1日 - 2020年9月30日 総長補佐
• 2020年10月12日 - 2022年3月31日 総長補佐
• 2022年4月1日 - 2024年3月31日 総長補佐
• 2020年10月12日 -  総長補佐（広報室担当）

委員歴

• 2022年 - 現在   日本動物学会   理事
• 2020年 - 現在   染色体学会   理事
• 2019年 - 現在   日本遺伝学会   編集委員
• 2019年 - 現在   日本遺伝学会   評議員（全国区）
• 2009年 - 2020年   染色体学会   評議員
• 2009年 - 2015年   日本遺伝学会   評議員（北海道地区)
• 2012年 - 2014年   日本動物学会   北海道支部会計幹事
• 2012年 - 2014年   日本動物学会   北海道支部札樽地区委員

メディア報道

• オトコとオンナ “性”のゆらぎのミステリー

  報道 : 2020年10月

  発行元・放送局 : HNK

  番組・新聞雑誌 : BSプレミアム「ヒューマニエンス〜40億年のたくらみ〜」

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• “男はつらい”ってホント？

  報道 : 2018年05月

  発行元・放送局 : HNK

  番組・新聞雑誌 : Eテレ「又吉直樹のヘウレーカ！」

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• ♂&♀はミステリー 性決定の不思議な世界

  報道 : 2015年10月

  発行元・放送局 : BSフジ

  番組・新聞雑誌 : ガリレオX

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• 沖縄やんばる 幻のオキナワトゲネズミ再発見

  報道 : 2009年04月

  発行元・放送局 : NHK

  番組・新聞雑誌 : ハイビジョン「プレミアム8 ワイルドライフ」
• Y染色体のミステリー

  報道 : 2009年02月

  発行元・放送局 : NHK

  番組・新聞雑誌 : ハイビジョン特集
• 男が消える？人類も消える？

  報道 : 2009年01月

  発行元・放送局 : NHK

  番組・新聞雑誌 : NHKスペシャル「女と男」

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