2002 Fiscal Year Annual Research Report

中枢神経系幹細胞の分化機構の解析

Research Project

Project/Area Number	12308042
Research Institution	Keio University
Principal Investigator	岡野栄之慶應義塾大学, 医学部, 教授 (60160694)
Keywords	gp130 / 神経幹細胞 / EGFのレセプター / Gab1 / 欠失変異体マウス / 長期維持機構 / オリゴデンドロサイト前駆細胞 / GFAP
Research Abstract	本研究では、gp130、そして神経幹細胞のmitogenであるEGFのレセプターの下流で働くシグナル伝達分子であり、MAP kinase経路の活性化に関与しているGab1の欠失変異体マウスにおける神経幹細胞の動態を含めた中枢神経系発生の異常を解析することによって、神経幹細胞の長期維持機構およびその本質に更にせまることを目的としている。 Gab1のホモ欠失変異体(gab^<-/->)は胎生16日目までに胎盤の発達異常により死亡してしまう故に、まず、胎生14日目の線条体における神経幹細胞の数をneurosphere法によって野生型と比較したところ、顕著な差は見られなかったが、EGFに対する反応性は消失していた。また、この時期に現れ始めるオリゴデンドロサイト前駆細胞の数の減少が見られたが、ニューロンの分化および生存に変化は見られなかった。一方、CNTFなどのgp130を介したシグナルによって正に制御されているGFAPの発現は、生後7日目のヘテロ欠失変異体(gab^<+/->)の脊髄で亢進していた。そのため、胎生14日目の脊髄の初代培養の系において、CNTF存在下で3日間培養を行ったところ、ヘテロ欠失変異体(gab^<+/->)ではGFAPの発現量が野生型に比べて亢進しており、CNTFに対する反応性が亢進していることが確認された。さらに成体においては、ヘテロ欠失変異体(gab^<+/->)の側脳室周囲に存在している神経幹細胞の数を、neurosphere法によって調べた結果、野生型に比べて約80%その数が増加していた。以上より、Gab1は(1)胎生期では主にオリゴデンドロサイト前駆細胞の増殖あるいは生存に必要であること、(2)GFAPの発現、つまりアストロサイトの分化においては抑制的に働くこと、(3)主に生後から成体に至るまでの、神経幹細胞の維持には負に作用していることが示唆された。

Research Products

(26 results)

All Other

All Publications (26 results)

[Publications] Iwai Y, Hirota Y, Ozaki K, Okano H, Takeichi M, Uemura T: "DN-cadherin-dependent synaptogenesis in the Drosophila visual system"Mol Cell Neurosci. 19. 375-388 (2002)
[Publications] Takasawa K-I, Kitagawa K, Yagita Y, Sasaki T, Tanaka S, Ohtsuki T, Miyata T, Okano H, Hori M, Matsumoto M: "Increased proliferation of neural progenitor cells but reduced survival newborn cells in the contralateral hippocampus after focal cerebral ischemia in rats"J Cerebral Blood Flow and Metabolism. 22. 299-307 (2002)
[Publications] Shibata M, Yamawaki T, Fukuuchi Y, Okano H, Miura M: "Up-reguration of Akt Phosphorylation Contributes to Improved Cell Viability in the Ischemic Penumbra at the Early Stage of Middle Cerebral Artery Occlusion in Mice"Brain Res. 942. 1-10 (2002)
[Publications] Shu H-J, Saito T, Watanabe H, Ito J-I, Takeda H, Okano H, Kawata S: "Expression of the Musashil gene encoding the RNA-binding protein in human hepatoma cell lines"Biochem. Biophys. Res. Com. 293. 150-154 (2002)
[Publications] Cuadrado, A., Garcia-Fernandez, L.F., Imai, T., Okano, H., Munoz, A: "Neural RNA-binding protein Musashi-1 modulates tau pre-mRNA alternative splicing and is regulated by thyroid hormone in the developing rat brain"Mol. Cell. Neurosci.. 20. 98-210 (2002)
[Publications] Shamloula, H.K., Mbogho, M.P., Pimentel, A.C., Chrzanowska-Lightowlers, Z.M.A., Hyatt, V., Okano, H., Venkatech.T.R: "rugose (rg), a Drosophila A kinase Anchor Protein (DAKAP), is required for required for retinal pattern formation and interacts genetically with multiple signaling pathways"Genetics. 161. 693-710 (2002)
[Publications] Yagita, Y., Kitagawa, K., Sasaki, T., Miyata, T., Okano H.Hori, T., Matsumoto, M: "Musashil and Nestin as useful markers of neuronal progenitor cells in rat hippocampus after forebrain ischemia"J. Neurosci. Res. 69. 750-756 (2002)
[Publications] Ogawa, Y., Sawamoto, K., Miyata, T., Miyao, S., Watanabe, M., Toyama, Y., Nakamura, M., Bregman, B.S., Koike, M., Uchiyama, Y., Okano, H.: "Transplantation of in vitro expanded fetal neural progenitor cells results in neurogenesis and functional recovery after spinal cord contusion injury in adult rats"J. Neurosci. Res. 69. 925-933 (2002)
[Publications] Murayama A, Matsuzaki Y, Kawaguti A, Shimazaki T, Okano H: "Flow cytometric analysis of stem cells in the developing and adult mouse brain"J. Neurosci. Res. 69. 837-847 (2002)
[Publications] Johansson CB, Lothian C, Molin M, Okano H, Lendahl U: "Nestin enhancer requirements for expression in normal and injured adult CNS"J. Neurosci. Res. 69. 784-794 (2002)
[Publications] Kanemura, Y., Mon, H., Kobayashi, S., Islam, 0., Kodama E., Yamamoto, A., Nakanishi Y., Arita, N., Yamasaki, M., Okano, H., Miyake, J., Hara, M.: "Evaluation of in-vitro proliferative activity of human fetal neural stem cell"J. Neurosci. Res. 69. 869-879 (2002)
[Publications] Kuranaga, E., Kanuka, H., Igaki, T., Sawamoto, K., Ichijo, H., Okano, H., Miura, M: "Reaper-mediated inhibition of DIAP1-induced DTRAF1degradation results in activation of JNK in Drosophila"Nat Cell Biol. 4. 705-710 (2002)
[Publications] Sakakibara, S., Nakamura, Y, Koike, M., Takano, H., Uchiyama, Y., Noda, T., Okano, H: "RNA-binding protein Musashi family, roles for CNS stem cells and a subpopulation of ependymal cells revealed by targeted disruption and antisense ablation"Proc. Natl. Acad. Sci. USA. 99. 15194-15199 (2002)
[Publications] Hirota Y., Sawamoto, K., Okano, H: "Tincar encodes a novel transmembrane protein expressed in the Tinman-expressing cardioblasts of Drosophila"Gene Expression Patterns. 2. 323-327 (2002)
[Publications] Okano, H., Yoshizaki, T., Shimazaki, T., Sawamoto, K: "Isolation and transplantation of Dopaminergic Neurons and Neural Stem Cells"Parkinson & Related Disorders. 9. 23-28 (2002)
[Publications] Okano, H., Imai, T., Okabe, M: "Musashi : a translational regulator of cell fates"J. Cell Sci. 115. 1355-1359 (2002)
[Publications] Okano, H: "The stem cell biology of the central nervous system"J. Neurosci. Res. 69. 698-707 (2002)
[Publications] Okano, H: "Neural stem cells: progression of basic research and perspective for clinical application"Keio J. Med. 51. 115-128 (2002)
[Publications] Potten, C.S., Booth C., Tudor, G.L., Booth, D., Brady, G., Hurley, P., Ashton, G., Clarke, G., Sakakibara, S., Okano.H.: "Identification of a putative intestinal stem cell marker and early lineage marker; Musashi1"Differentiation. 71. 28-41 (2003)
[Publications] Kayahara, T., Sawada, M., Takaishi, S., Fukui, H., Seno, H., Fukuzawa, H., Suzuki, K., Hiai, H., Kageyama, R., Chiba, T.: "Candidate markers for stem and early progenitor cells, Musashi-1 and Hes1, are expressed in crypt base columnar cells of mouse small intestine"FEBS Letter. 535. 131-135 (2003)
[Publications] Tonchev, A.B., Zhao, L., Yamashima, T., Okano, H: "Differential proliferative response in the postischemic hippocampus, temporal cortex and olfactory bulb of adult primates"Glia. (in press). (2003)
[Publications] Tonchev, A.B., Yamashima, T., Zhao, L., Okano, H.J., Okano, H: "Increased proliferation of neural progenitors and neurogenesis in the postischemic brain of young adult primates"Mol. Cell. Neurosci. (in press). (2003)
[Publications] Sasaki, T., Kitagawa, K., Sugimura, S., Omura-Matsuoka, E., Tanaka, S., Yagita, Y., Okano, H: "Matsumoto, M. and Hori, M. : Implication of Cyclooxygenase-2 on enhanced neurogenesis in the adult hippocampus after ischemia"J. Neurosci. Res. (in press). (2003)
[Publications] Nakamura, Y, Yamamoto, M., Oda, B., Yamamoto, A., Kanemura, Y., Hara, M., Suzuki, A., Yamasaki, M., Okano, H: "Expression of tubulin beta II in neural stem/progenitor cells and radial fibers during human fetal brain development"Lab Invest. (in press). (2003)
[Publications] Okano H, Ogawa, Y, Nakamura, M., Kaneko, S., Iwanami, A., Toyama, A.: "Transplantation of neural stem cells into the spinal cord after injury"Seminar in Cell & Devlopmental Biol. (in press). (2003)
[Publications] Sawamoto K, Okano H: "Recent Res. Devel. Mol. Cell Biol."Research Signpost. 661 (2002)

2002 Fiscal Year Annual Research Report

中枢神経系幹細胞の分化機構の解析

Principal Investigator

岡野 栄之 慶應義塾大学, 医学部, 教授 (60160694)

Research Products

[Publications] Iwai Y, Hirota Y, Ozaki K, Okano H, Takeichi M, Uemura T: "DN-cadherin-dependent synaptogenesis in the Drosophila visual system"Mol Cell Neurosci. 19. 375-388 (2002)

[Publications] Shibata M, Yamawaki T, Fukuuchi Y, Okano H, Miura M: "Up-reguration of Akt Phosphorylation Contributes to Improved Cell Viability in the Ischemic Penumbra at the Early Stage of Middle Cerebral Artery Occlusion in Mice"Brain Res. 942. 1-10 (2002)

[Publications] Shu H-J, Saito T, Watanabe H, Ito J-I, Takeda H, Okano H, Kawata S: "Expression of the Musashil gene encoding the RNA-binding protein in human hepatoma cell lines"Biochem. Biophys. Res. Com. 293. 150-154 (2002)

[Publications] Cuadrado, A., Garcia-Fernandez, L.F., Imai, T., Okano, H., Munoz, A: "Neural RNA-binding protein Musashi-1 modulates tau pre-mRNA alternative splicing and is regulated by thyroid hormone in the developing rat brain"Mol. Cell. Neurosci.. 20. 98-210 (2002)

[Publications] Yagita, Y., Kitagawa, K., Sasaki, T., Miyata, T., Okano H.Hori, T., Matsumoto, M: "Musashil and Nestin as useful markers of neuronal progenitor cells in rat hippocampus after forebrain ischemia"J. Neurosci. Res. 69. 750-756 (2002)

[Publications] Murayama A, Matsuzaki Y, Kawaguti A, Shimazaki T, Okano H: "Flow cytometric analysis of stem cells in the developing and adult mouse brain"J. Neurosci. Res. 69. 837-847 (2002)

[Publications] Johansson CB, Lothian C, Molin M, Okano H, Lendahl U: "Nestin enhancer requirements for expression in normal and injured adult CNS"J. Neurosci. Res. 69. 784-794 (2002)

[Publications] Kanemura, Y., Mon, H., Kobayashi, S., Islam, 0., Kodama E., Yamamoto, A., Nakanishi Y., Arita, N., Yamasaki, M., Okano, H., Miyake, J., Hara, M.: "Evaluation of in-vitro proliferative activity of human fetal neural stem cell"J. Neurosci. Res. 69. 869-879 (2002)

[Publications] Kuranaga, E., Kanuka, H., Igaki, T., Sawamoto, K., Ichijo, H., Okano, H., Miura, M: "Reaper-mediated inhibition of DIAP1-induced DTRAF1degradation results in activation of JNK in Drosophila"Nat Cell Biol. 4. 705-710 (2002)

[Publications] Hirota Y., Sawamoto, K., Okano, H: "Tincar encodes a novel transmembrane protein expressed in the Tinman-expressing cardioblasts of Drosophila"Gene Expression Patterns. 2. 323-327 (2002)

[Publications] Okano, H., Yoshizaki, T., Shimazaki, T., Sawamoto, K: "Isolation and transplantation of Dopaminergic Neurons and Neural Stem Cells"Parkinson & Related Disorders. 9. 23-28 (2002)

[Publications] Okano, H., Imai, T., Okabe, M: "Musashi : a translational regulator of cell fates"J. Cell Sci. 115. 1355-1359 (2002)

[Publications] Okano, H: "The stem cell biology of the central nervous system"J. Neurosci. Res. 69. 698-707 (2002)

[Publications] Okano, H: "Neural stem cells: progression of basic research and perspective for clinical application"Keio J. Med. 51. 115-128 (2002)

[Publications] Potten, C.S., Booth C., Tudor, G.L., Booth, D., Brady, G., Hurley, P., Ashton, G., Clarke, G., Sakakibara, S., Okano.H.: "Identification of a putative intestinal stem cell marker and early lineage marker; Musashi1"Differentiation. 71. 28-41 (2003)

[Publications] Tonchev, A.B., Zhao, L., Yamashima, T., Okano, H: "Differential proliferative response in the postischemic hippocampus, temporal cortex and olfactory bulb of adult primates"Glia. (in press). (2003)

[Publications] Tonchev, A.B., Yamashima, T., Zhao, L., Okano, H.J., Okano, H: "Increased proliferation of neural progenitors and neurogenesis in the postischemic brain of young adult primates"Mol. Cell. Neurosci. (in press). (2003)

[Publications] Sasaki, T., Kitagawa, K., Sugimura, S., Omura-Matsuoka, E., Tanaka, S., Yagita, Y., Okano, H: "Matsumoto, M. and Hori, M. : Implication of Cyclooxygenase-2 on enhanced neurogenesis in the adult hippocampus after ischemia"J. Neurosci. Res. (in press). (2003)

[Publications] Nakamura, Y, Yamamoto, M., Oda, B., Yamamoto, A., Kanemura, Y., Hara, M., Suzuki, A., Yamasaki, M., Okano, H: "Expression of tubulin beta II in neural stem/progenitor cells and radial fibers during human fetal brain development"Lab Invest. (in press). (2003)

[Publications] Okano H, Ogawa, Y, Nakamura, M., Kaneko, S., Iwanami, A., Toyama, A.: "Transplantation of neural stem cells into the spinal cord after injury"Seminar in Cell & Devlopmental Biol. (in press). (2003)

[Publications] Sawamoto K, Okano H: "Recent Res. Devel. Mol. Cell Biol."Research Signpost. 661 (2002)

岡野栄之慶應義塾大学, 医学部, 教授 (60160694)