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依馬 正次

所属部署名動物生命科学研究センター
職名教授
 
 
更新日: 17/11/17 09:58

研究者基本情報

氏名

    依馬 正次

所属(マスタ)

  • 動物生命科学研究センター 教授

学歴

  • 1993年04月- 1996年03月東北大学大学院 理学研究科 博士課程
  • 1991年04月- 1993年03月東北大学大学院 理学研究科 修士課程
  • 1987年04月- 1991年03月東北大学 理学部 化学第二学科

学位

  • 博士(理学)(東北大学)

所属学協会

    日本生化学会 , 日本分子生物学会 , 日本癌学会 , 日本再生医療学会 , 日本血管生物医学会

委員歴

  • 2014年- 現在日本生化学会 評議員
  • 2014年- 現在日本血管生物医学会 評議員

経歴

  • 2013年09月- 現在滋賀医科大学(教授)
  • 2010年10月- 2015年03月さきがけ・JST(研究員)
  • 2013年07月- 2014年08月筑波大学(准教授)
  • 筑波大学 医学医療系 (講師)
  • 2002年- 筑波大学基礎医学系講師
  • 1998年- 2002年 マウントサイナイ病院、ポスドク
  • 1996年- 1998年 東北大学大学院理学研究科、ポスドク

研究活動情報

研究分野

  • 生物科学 / 発生生物学

書籍等出版物

  • 酸素分圧による遺伝子発現制御機構
    共立出版酸化ストレス・レドックスの生化学 2000年
  • Flk1 expression marks broad spectrum of mesodermal progenitors in the early mouse embryo
    第25回日本分子生物学会年会 2003年
  • マウス成体型造血の不均一性
    第25回日本分子生物学会年会 2003年
  • Cell fate decisions regulated by VEGF and its receptors in multipotent mesodermal progenitors in the early embryo
    International meeting of Differentiation 2004年

MISC

  • Klf5 maintains the balance of primitive endoderm versus epiblast specification during mouse embryonic development by suppression of Fgf4
    Takuya Azami, Takuya Azami, Tsuyoshi Waku, Ken Matsumoto, Hyojung Jeon, Masafumi Muratani, Akihiro Kawashima, Jun Yanagisawa, Jun Yanagisawa, Ichiro Manabe, Ryozo Nagai, Tilo Kunath, Tomonori Nakamura, Tomonori Nakamura, Kazuki Kurimoto, Kazuki Kurimoto, Mitinori Saitou, Mitinori Saitou, Mitinori Saitou, Mitinori Saitou, Satoru Takahashi, Satoru Takahashi, Satoru Takahashi, Masatsugu Ema, Masatsugu Ema
    Development (Cambridge) 144 3706-3718 2017年10月
    © 2017. Published by The Company of Biologists Ltd. The inner cell mass of the mouse blastocyst gives rise to the pluripotent epiblast (EPI), which forms the embryo proper, and the primitive endoderm (PrE), which forms extra-embryonic yolk sac tissues. All inner cells coexpress lineage markers such as Nanog and Gata6 at embryonic day (E) 3.25, and the EPI and PrE precursor cells eventually segregate to exclusively express Nanog and Gata6, respectively. Fibroblast growth factor (FGF)–extracellular signal-regulated kinase (ERK) signalling is involved in segregation of the EPI and PrE lineages; however, the mechanism involved in Fgf4 regulation is poorly understood. Here, we identified Klf5 as an upstream repressor of Fgf4. Fgf4 was markedly upregulated in Klf5 knockout (KO) embryos at E3.0, and was downregulated in embryos overexpressing Klf5. Furthermore, Klf5 KO and overexpressing blastocysts showed skewed lineage specification phenotypes, similar to FGF4-treated preimplantation embryos and Fgf4 KO embryos, respectively. Inhibitors of the FGF receptor (Fgfr) and ERK pathways reversed the skewed lineage specification of Klf5 KO blastocysts. These data demonstrate that Klf5 suppresses Fgf4-Fgfr- ERK signalling, thus preventing precocious activation of the PrE specification programme.
  • Prox1-GFP/Flt1-DsRed transgenic mice: an animal model for simultaneous live imaging of angiogenesis and lymphangiogenesis
    Wei Zhong, Wei Zhong, Wei Zhong, Xinbo Gao, Shuangyong Wang, Kyuyeon Han, Masatsugu Ema, Susanne Adams, Ralf H. Adams, Mark I. Rosenblatt, Jin Hong Chang, Dimitri T. Azar
    Angiogenesis 1-18 2017年08月
    © 2017 Springer Science+Business Media B.V. The roles of angiogenesis in development, health, and disease have been studied extensively; however, the studies related to lymphatic system are limited due to the difficulty in observing colorless lymphatic vessels. But recently, with the improved technique, the relative importance of the lymphatic system is just being revealed. We bred transgenic mice in which lymphatic endothelial cells express GFP (Prox1-GFP) with mice in which vascular endothelial cells express DsRed (Flt1-DsRed) to generate Prox1-GFP/Flt1-DsRed (PGFD) mice. The inherent fluorescence of blood and lymphatic vessels allows for direct visualization of blood and lymphatic vessels in various organs via confocal and two-photon microscopy and the formation, branching, and regression of both vessel types in the same live mouse cornea throughout an experimental time course. PGFD mice were bred with CDh5CreERT2 and VEGFR2lox knockout mice to examine specific knockouts. These studies showed a novel role for vascular endothelial cell VEGFR2 in regulating VEGFC-induced corneal lymphangiogenesis. Conditional deletion of vascular endothelial VEGFR2 abolished VEGFA- and VEGFC-induced corneal lymphangiogenesis. These results demonstrate the potential use of the PGFD mouse as a powerful animal model for studying angiogenesis and lymphangiogenesis.
  • Tissue Myeloid Progenitors Differentiate into Pericytes through TGF-β Signaling in Developing Skin Vasculature
    Tomoko Yamazaki, Ani Nalbandian, Yutaka Uchida, Wenling Li, Thomas D. Arnold, Yoshiaki Kubota, Seiji Yamamoto, Masatsugu Ema, Yoh suke Mukouyama
    Cell Reports 18 2991-3004 2017年03月
    © 2017 Mural cells (pericytes and vascular smooth muscle cells) are essential for the regulation of vascular networks and maintenance of vascular integrity, but their origins are diverse in different tissues and not known in the organs that arise from the ectoderm, such as skin. Here, we show that tissue-localized myeloid progenitors contribute to pericyte development in embryonic skin vasculature. A series of in vivo fate-mapping experiments indicates that tissue myeloid progenitors differentiate into pericytes. Furthermore, depletion of tissue myeloid cells and their progenitors in PU.1 (also known as Spi1) mutants results in defective pericyte development. Fluorescence-activated cell sorting (FACS)-isolated myeloid cells and their progenitors from embryonic skin differentiate into pericytes in culture. At the molecular level, transforming growth factor-β (TGF-β) induces pericyte differentiation in culture. Furthermore, type 2 TGF-β receptor (Tgfbr2) mutants exhibit deficient pericyte development in skin vasculature. Combined, these data suggest that pericytes differentiate from tissue myeloid progenitors in the skin vasculature through TGF-β signaling.
  • Visualization of the epiblast and visceral endodermal cells using Fgf5-P2A-venus BAC transgenic mice and epiblast stem cells
    Le Tran Phuc Khoa, Takuya Azami, Takuya Azami, Tomoyuki Tsukiyama, Jun Matsushita, Setsuko Tsukiyama-Fujii, Satoru Takahashi, Satoru Takahashi, Satoru Takahashi, Masatsugu Ema, Masatsugu Ema
    PLoS ONE 11 2016年07月
    © 2016 Khoa et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.Fibroblast growth factor 5 (Fgf5) has been widely used as a marker for the epiblast in the postimplantation embryo and epiblast stem cells (mEpiSCs) in the mouse, making it valuable for study of differentiation of various tissues and epiblast cells in vivo and in vitro. Here, we report for the first time the generation of Fgf5-P2A-Venus BAC transgenic (Tg) mice and show that the BAC Tg can recapitulate endogenous Fgf5 expression in epiblast and visceral endodermal cells of E6.5 and 7.5 embryos. We also show that Fgf5-P2A-Venus BAC Tg mEpiSCs in the undifferentiated state expressed abundant Venus, and upon reprogramming into naïve state, Venus was suppressed. Furthermore, while most Tg mEpiSCs expressed Venus abundantly, surprisingly the Tg mEpiSCs contained a minor subpopulation of Venus-negative cells that were capable of conversion to Venus-positive cells, indicating that even Fgf5 expression shows dynamic heterogeneity in mEpiSCs. Taken together, Fgf5-P2A-Venus BAC Tg mice and mEpiSCs generated in this study will be useful for developmental biology as well as stem cell biology research.
  • Generation of transgenic cynomolgus monkeys that express green fluorescent protein throughout the whole body
    Yasunari Seita, Yasunari Seita, Tomoyuki Tsukiyama, Chizuru Iwatani, Chizuru Iwatani, Hideaki Tsuchiya, Jun Matsushita, Jun Matsushita, Takuya Azami, Junko Okahara, Shinichiro Nakamura, Yoshitaka Hayashi, Seiji Hitoshi, Yasushi Itoh, Takeshi Imamura, Masaki Nishimura, Ikuo Tooyama, Hiroyuki Miyoshi, Mitinori Saitou, Mitinori Saitou, Mitinori Saitou, Mitinori Saitou, Kazumasa Ogasawara, Erika Sasaki, Masatsugu Ema, Masatsugu Ema
    Scientific Reports 6 2016年04月
    © 2016, Nature Publishing Group. All rights reserved.Nonhuman primates are valuable for human disease modelling, because rodents poorly recapitulate some human diseases such as Parkinson's disease and Alzheimer's disease amongst others. Here, we report for the first time, the generation of green fluorescent protein (GFP) transgenic cynomolgus monkeys by lentivirus infection. Our data show that the use of a human cytomegalovirus immediate-early enhancer and chicken beta actin promoter (CAG) directed the ubiquitous expression of the transgene in cynomolgus monkeys. We also found that injection into mature oocytes before fertilization achieved homogenous expression of GFP in each tissue, including the amnion, and fibroblasts, whereas injection into fertilized oocytes generated a transgenic cynomolgus monkey with mosaic GFP expression. Thus, the injection timing was important to create transgenic cynomolgus monkeys that expressed GFP homogenously in each of the various tissues. The strategy established in this work will be useful for the generation of transgenic cynomolgus monkeys for transplantation studies as well as biomedical research.
  • Comprehensive identification of Krüppel-like factor family members contributing to the self-renewal of mouse embryonic stem cells and cellular reprogramming
    Hyojung Jeon, Tsuyoshi Waku, Takuya Azami, Le Tran Phuc Khoa, Jun Yanagisawa, Jun Yanagisawa, Satoru Takahashi, Satoru Takahashi, Satoru Takahashi, Masatsugu Ema, Masatsugu Ema
    PLoS ONE 11 2016年03月
    © 2016 Jeon et al.This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.Pluripotency is maintained in mouse embryonic stem (ES) cells and is induced from somatic cells by the activation of appropriate transcriptional regulatory networks. Krüppel-like factor gene family members, such as Klf2, Klf4 and Klf5, have important roles in maintaining the undifferentiated state of mouse ES cells as well as in cellular reprogramming, yet it is not known whether otherKlf family members exert self-renewal and reprogramming functions when overexpressed. In this study, we examined whether overexpression of any representative Klf family member, such as Klf1-Klf10, would be sufficient for the self-renewal of mouse ES cells. We found that only Klf2, Klf4, and Klf5 produced leukemia inhibitory factor (LIF)-independent self-renewal, although most KLF proteins, if not all, have the ability to occupy the regulatory regions of Nanog, a critical Klf target gene. We also examined whether overexpression of any of Klf1-Klf10 would be sufficient to convert epiblast stem cells into a naïve pluripotent state and found that Klf5 had such reprogramming ability, in addition to Klf2 and Klf4. We also delineated the functional domains of the Klf2 protein for LIF-independent self-renewal and reprogramming. Interestingly, we found that both the N-terminal transcriptional activation and C-terminal zinc finger domains were indispensable for this activity. Taken together, our comprehensive analysis provides new insight into the contribution of Klf family members to mouse ES self-renewal and cellular reprogramming.
  • Developmental regression of hyaloid vasculature is triggered by neurons
    Yusuke Yoshikawa, Yusuke Yoshikawa, Toru Yamada, Toru Yamada, Ikue Tai-Nagara, Keisuke Okabe, Keisuke Okabe, Yuko Kitagawa, Masatsugu Ema, Yoshiaki Kubota
    Journal of Experimental Medicine 213 1175-1183 2016年01月
    © 2016 Yoshikawa et al.Vascular development involves not only vascular growth, but also regression of transient or unnecessary vessels. Hyaloid vasculature is the temporary circulatory system in fetal eyes, which spontaneously degenerates when the retinal blood vessels start to grow. Failure of the hyaloid vessels to regress leads to disease in humans, persistent hyperplastic primary vitreous, which causes severe intraocular hemorrhage and impairs visual function. However, the mechanism underlying the endogenous program that mediates spontaneous regression of the hyaloid vessels is not well understood. In this study, we identify a robust switch triggering this program directed by neurons in mice. Marked up-regulation of vascular endothelial growth factor (VEGF) receptor 2 (VEG FR2) occurs in retinal neurons just after birth via distal-multipotent-mesodermal enhancer, a hemangioblast- specific enhancer of VEG FR2. Genetic deletion of neuronal VEG FR2 interrupts this program, resulting in massive hyaloid vessels that persist even during late postnatal days. This abnormality is caused by excessive VEGF proteins in the vitreous cavity as a result of impairment in the neuronal sequestration of VEGF. Collectively, our data indicate that neurons trigger transition from the fetal to the postnatal circulatory systems in the retina.
  • Arf6 regulates tumour angiogenesis and growth through HGF-induced endothelial β1 integrin recycling
    Tsunaki Hongu, Yuji Funakoshi, Shigetomo Fukuhara, Teruhiko Suzuki, Teruhiko Suzuki, Susumu Sakimoto, Susumu Sakimoto, Nobuyuki Takakura, Masatsugu Ema, Masatsugu Ema, Satoru Takahashi, Susumu Itoh, Susumu Itoh, Mitsuyasu Kato, Hiroshi Hasegawa, Hiroshi Hasegawa, Naoki Mochizuki, Yasunori Kanaho
    Nature Communications 6 2015年08月
    © 2015 Macmillan Publishers Limited. All rights reserved.Anti-angiogenic drugs targeting vascular endothelial cell growth factor receptor have provided modest clinical benefit, in part, owing to the actions of additional angiogenic factors that stimulate tumour neoangiogenesis in parallel. To overcome this redundancy, approaches targeting these other signalling pathways are required. Here we show, using endothelial cell-targeted mice, that the small GTPase Arf6 is required for hepatocyte growth factor (HGF)-induced tumour neoangiogenesis and growth. Arf6 deletion from endothelial cells abolishes HGF-stimulated β1 integrin recycling. Pharmacological inhibition of the Arf6 guanine nucleotide exchange factor (GEF) Grp1 efficiently suppresses tumour vascularization and growth. Grp1 as well as other Arf6 GEFs, such as GEP100, EFA6B and EFA6D, regulates HGF-stimulated β1 integrin recycling. These findings provide insight into the mechanism of HGF-induced tumour angiogenesis and offer the possibility that targeting the HGF-activated Arf6 signalling pathway may synergize with existing anti-angiogenic drugs to improve clinical outcomes.
  • Functional compensation between Myc and PI3K signaling supports self-renewal of embryonic stem cells
    Tomoaki Hishida, Tomoaki Hishida, Tomoaki Hishida, Yutaka Nakachi, Yutaka Nakachi, Yosuke Mizuno, Miyuki Katano, Yasushi Okazaki, Yasushi Okazaki, Yasushi Okazaki, Masatsugu Ema, Satoru Takahashi, Satoru Takahashi, Masataka Hirasaki, Ayumu Suzuki, Atsushi Ueda, Masazumi Nishimoto, Yuriko Hishida-Nozaki, Eric Vazquez-Ferrer, Ignacio Sancho-Martinez, Juan Carlos Izpisua Belmonte, Akihiko Okuda, Akihiko Okuda
    Stem Cells 33 713-725 2015年03月
    © 2014 AlphaMed Press.c-Myc and phosphatidylinositol 3-OH kinase (PI3K) both participate in diverse cellular processes, including cell cycle control and tumorigenic transformation. They also contribute to preserving embryonic stem cell (ESC) characteristics. However, in spite of the vast knowledge, the molecular relationship between c-Myc and PI3K in ESCs is not known. Herein, we demonstrate that c-Myc and PI3K function cooperatively but independently to support ESC self-renewal when murine ESCs are cultured under conventional culture condition. Interestingly, culture of ESCs in 2i-condition including a GSK3β and MEK inhibitor renders both PI3K and Myc signaling dispensable for the maintenance of pluripotent properties. These results suggest that the requirement for an oncogenic proliferation-dependent mechanism sustained by Myc and PI3K is context dependent and that the 2i-condition liberates ESCs from the dependence of this mechanism. Stem Cells 2015;33:713-725
  • Forced expression of nanog or esrrb preserves the ESC status in the absence of nucleostemin expression
    Miyuki Katano, Masatsugu Ema, Yutaka Nakachi, Yutaka Nakachi, Yosuke Mizuno, Masataka Hirasaki, Ayumu Suzuki, Atsushi Ueda, Masazumi Nishimoto, Masazumi Nishimoto, Satoru Takahashi, Yasushi Okazaki, Yasushi Okazaki, Akihiko Okuda, Akihiko Okuda
    Stem Cells 33 1089-1101 2015年01月
    © 2015 Alpha Med Press.Nucleostemin (NS) is a nucleolar GTP-binding protein that is involved in a plethora of functions including ribosomal biogenesis and maintenance of telomere integrity. In addition to its expression in cancerous cells, the NS gene is expressed in stem cells including embryonic stem cells (ESCs). Previous knockdown and knockout studies have demonstrated that NS is important to preserve the self-renewality and high expression levels of pluripotency marker genes in ESCs. Here, we found that forced expression of Nanog or Esrrb, but not other pluripotency factors, resulted in the dispensability of NS expression in ESCs. However, the detrimental phenotypes of ESCs associated with ablation of NS expression were not mitigated by forced expression of Rad51 or a nucleolar localization-defective NS mutant that counteracts the damage associated with loss of NS expression in other NS-expressing cells such as neural stem/progenitor cells. Thus, our results indicate that NS participates in preservation of the viability and integrity of ESCs, which is distinct from that in other NS-expressing cells.
  • Combined administration of BMP-2 and HGF facilitate bone regeneration through angiogenic mechanisms
    Tomoyuki Masuda, Tomoyuki Masuda, Keishi Otsu, Mika Kumakami-Sakano, Naoki Fujiwara, Masatsugu Ema, Jiro Hitomi, Yoshiki Sugiyama, Hidemitsu Harada
    Journal of Hard Tissue Biology 24 7-16 2015年01月
    © 2015 The Hard Tissue Biology Network Association Printed in Japan, All rights reserved.Promotion of bone repair contributes to the shortening of the treatment period and improvement of the therapeutic effect in large bone defects. The purpose of the present study was to determine the optimal condition for the administration of growth factors to facilitate bone repair and explore the significance of angiogenesis in bone regeneration in vivo. Critical-size calvarial defects were created in the parietal bones of adult ddY mice, and the defects were treated with gelatin sponges impregnated with growth factors BMP-2, FGF-2, and HGF, alone or in combination. The bone regeneration at 1, 2, 3, and 4 weeks was evaluated using micro-computed tomography and histological observation. Critical-size calvarial defects were also created in Flk1-GFP mice to analyze angiogenesis in the early phase of bone repair. The proliferation of Flk1-GFP positive endothelial cells during bone repair was quantitatively evaluated using immunohistochemistry for Ki67. Treatment with a combination of BMP-2 and HGF (BMP-2+HGF) significantly induced more new bone formation within the bone defects compared with the other groups. Compared with BMP-2 alone, bone regeneration was rapidly increased by BMP-2+HGF from 3 weeks of the beginning of the treatment. The number of FLK1-GFP and Ki67 double positive cells in the defect areas of mice administered BMP-2+HGF at 1 week was larger than that in those administered BMP-2 alone. These results indicate that the combination of BMP-2 and HGF was markedly efficient in bone regeneration due to the promotion of angiogenesis in the early phase of bone repair.
  • Osteogenic capillaries orchestrate growth plate-independent ossification of the malleus
    Koichi Matsuo, Yukiko Kuroda, Nobuhito Nango, Kouji Shimoda, Yoshiaki Kubota, Masatsugu Ema, Latifa Bakiri, Erwin F. Wagner, Yoshihiro Takeda, Wataru Yashiro, Atsushi Momose
    Development (Cambridge) 142 3912-3920 2015年01月
    © 2015. Published by The Company of Biologists Ltd.Endochondral ossification is a developmental process by which cartilage is replaced by bone. Terminally differentiated hypertrophic chondrocytes are calcified, vascularized, and removed by chondroclasts before bone matrix is laid down by osteoblasts. In mammals, the malleus is one of three auditory ossicles that transmit vibrations of the tympanic membrane to the inner ear. The malleus is formed from a cartilaginous precursor without growth plate involvement, but little is known about how bones of this type undergo endochondral ossification. Here, we demonstrate that in the processus brevis of the malleus, clusters of osteoblasts surrounding the capillary loop produce bone matrix, causing the volume of the capillary lumen to decrease rapidly in post-weaning mice. Synchrotron X-ray tomographic microscopy revealed a concentric, cylindrical arrangement of osteocyte lacunae along capillaries, indicative of pericapillary bone formation. Moreover, we report that overexpression of Fosl1, which encodes a component of the AP-1 transcription factor complex, in osteoblasts significantly blocked malleal capillary narrowing. These data suggest that osteoblast/endothelial cell interactions control growth plate-free endochondral ossification through ‘osteogenic capillaries’ in a Fosl1-regulated manner.
  • Neurons limit angiogenesis by titrating VEGF in retina
    Keisuke Okabe, Keisuke Okabe, Sakiko Kobayashi, Toru Yamada, Toru Yamada, Toshihide Kurihara, Ikue Tai-Nagara, Takeshi Miyamoto, Yoh Suke Mukouyama, Thomas N. Sato, Thomas N. Sato, Thomas N. Sato, Thomas N. Sato, Thomas N. Sato, Toshio Suda, Masatsugu Ema, Yoshiaki Kubota
    Cell 159 584-596 2014年10月
    © 2014 Elsevier Inc.Vascular and nervous systems, two major networks in mammalian bodies, show a high degree of anatomical parallelism and functional crosstalk. During development, neurons guide and attract blood vessels, and consequently this parallelism is established. Here, we identified a noncanonical neurovascular interaction in eye development and disease. VEGFR2, a critical endothelial receptor for VEGF, was more abundantly expressed in retinal neurons than in endothelial cells, including endothelial tip cells. Genetic deletion of VEGFR2 in neurons caused misdirected angiogenesis toward neurons, resulting in abnormally increased vascular density around neurons. Further genetic experiments revealed that this misdirected angiogenesis was attributable to an excessive amount of VEGF protein around neurons caused by insufficient engulfment of VEGF by VEGFR2-deficient neurons. Moreover, absence of neuronal VEGFR2 caused misdirected regenerative angiogenesis in ischemic retinopathy. Thus, this study revealed neurovascular crosstalk and unprecedented cellular regulation of VEGF: retinal neurons titrate VEGF to limit neuronal vascularization. PaperFlick

受賞

  • 1999年 生化学会奨励賞

競争的資金

  • 血液、血管、幹細胞の発生学的研究