2003 年度実績報告書

中枢神経系幹細胞の分化機構の解析

研究課題

研究課題/領域番号	12308042
研究機関	慶應義塾大学
研究代表者	岡野栄之慶應義塾大学, 医学部, 教授 (60160694)
キーワード	神経幹細胞 / モノクローナル抗体 / ephrin B1 / 放射状グリア細胞 / M6a / ニューロン / 神経発生 / 細胞分化
研究概要	神経発生過程における神経幹細胞の分化機構を明らかにすることを目的として、分化過程における特定の細胞を生きたまま同定することのできる抗体を作成した。胎生11日目のマウス胎仔期の終脳の細胞をラットに免疫し、約7000のハイブリドーマをクローン化した。まずFACSによって細胞表面を認識する抗体を選別し、次に胎仔脳の切片の免疫組織化学を行い、興味深い染色パターンを示す抗体を産生するクローンをスクリーニングした。プロトプラストフュージョン法による発現クローニングによって、これらのうち二つの抗体の抗原遺伝子を決定したところ、ephrinB1とM6aであることが明らかになった。 Ephrinは、受容体型チロシンキナーゼEphの膜結合型リガンドであり、軸索走行の制御など様々な生物現象に関与することが知られている。今回得られた抗体は、EphrinB1を特異的に認識し、他のEphrinBサブファミリーを認識しないことを確認した。本抗体を用いた免疫組織化学的解析によってEphrinB1は放射状グリアに強く発現していることが明らかになった。放射状グリア細胞には、ニューロンの移動の足場としての役割の他、神経幹細胞様の性質を有することが知られており、これらの細胞生物学的性質にEphrinB1の機能が関与している可能性が考えられる。さらに本抗体は、神経幹細胞のセルソーターによる分離にも役立つ可能性がある。 M6aは、PLP/DM20ファミリーに属する脂質蛋白質である。本研究で得られた抗体は、同じファミリーのメンバーであるPLPとは反応しないことを確認した。免疫組織化学的な解析によって、M6aは胎児脳における中間層に存在するニューロン様細胞に発現することを見いだした。

研究成果

(24件)

すべてその他

すべて文献書誌 (24件)

[文献書誌] Tonchev AB, Okano, H, et al.: "Proliferation of neural and neuronal progenitors after global brain ischemia in young adult macaque monkeys."Mol Cell Neurosci. 23. 292-301 (2003)
[文献書誌] Kayahara T, Okano H, et al.: "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)
[文献書誌] Sasaki T, Okano H, et al.: "Implication of Cyclooxygenase-2 on Enhanced Proliferation of Neural Progenitor Cells in the Adult Mouse Hippocampus After Ischemia."J Neurosci Res. 72. 461-471 (2003)
[文献書誌] Nakamura Y, Okano H, et al.: "Expression of tubulin beta II in neural stem/progenitor cells and radial fibers during human fetal brain development"Lab Invest. 83. 479-489 (2003)
[文献書誌] Yuasa Y, Okano H, et al.: "Drosophila homeodomain protein REPO controls glial differentiation by cooperating with ETS and BTB transcription factors."Dovelopment. 130. 2419-2428 (2003)
[文献書誌] Uchida K, Okano H, et al.: "Grafted swine neuroepithelial stem cells can form myelinated axons and both efferent and afferent synapses with xenogeneic rat neurons."J Neurosci Res. 72. 661-669 (2003)
[文献書誌] Kokuzawa J, Okano H, et al.: "Hepatocyte growth factor promotes proliferation and neural differentiation of neural stem cells from mouse embryos."Mol Cell Neurosci. 24. 190-197 (2003)
[文献書誌] Ishizuya-Oka A, Okano H, et al.: "Thyroid hormone-upregulated expression of Musashi-1 is specific for progenitor cells of the adult epithelium during amphibian gastrointestinal remodeling."J Cell Sci. 116. 3157-3164 (2003)
[文献書誌] Kanuka H, Okano H, et al.: "Cytosol-endoplasmic reticulum interplay by Sec61alpha translocon in polyglutamine-mediated neurotoxicity in Drosophila."Proc Natl Acad Sai USA. 100. 11723-11728 (2003)
[文献書誌] Baker H, Okano H, et al.: "Cortical and striatal expression of tyrosine hydroxylase mRNA in neonatal and adult mice."Cell Mol Neurobiol. 23. 503-518 (2003)
[文献書誌] Tamaki T, Okano H, et al.: "Growth and differentiation potential of main- and side-population cells derived from murine skeletal muscle."Exp Cell Res. 291. 83-90 (2003)
[文献書誌] Miyanomori Y, Okano H, et al.: "Origin of figher affinity to RNA of the N-terminal RNA-binding domain than that of the C-terminal one of a mouse neural protein, musashi1, as revealed by comparison of their structures, modes of interaction, surface electrostatic potentials, and backbone dynamics."J Biol Chem. 278. 41309-41315 (2003)
[文献書誌] Yoshida T, Okano, H: "Distinct expression patterns of splicing isoforms of mNumb in the endocrine lineage of developing pancreas."Differentiation. 71. 486-495 (2003)
[文献書誌] Hu QD, Okano H, et al.: "F3/Contactin acts as a functional ligand for Notch during oligodendrocyte maturation."Cell. 535. 131-135 (2003)
[文献書誌] Matsuzaki Y, Kinjo K, Mulligan RC, Okano H et al.: "Drosophila homeodomain protein REPO controls glial differentiation by cooperating with ETS and BTB transcription factors."Development. 130. 2419-2428 (2003)
[文献書誌] Uchida K, Okano H, et al.: "Grafted swine neuroepithelial stem cells can form myelinated axons and both efferent and afferent synapses with xenogeneic rat neurons."J Neurosci Res. 72. 661-669 (2003)
[文献書誌] Okada S, Okano H, et al.: "Implication of Cyclooxygenase-2 on Enhanced Proliferation of Neural Progenitor Cells in the Adult Mouse Hippocampus After Ischemia."J Neurosci Res. (In press). (2004)
[文献書誌] Yamashima T, Okano H et al.: "Vascular adventitia generates neuronal progenitors in monkey hippocampus after ischemia."Hippocampus. (In press). (2004)
[文献書誌] Ieda M, Okano H, et al.: "Distinct expression patterns of splicing isoforms of mNumb in the endocrine lineage of developing pancreas."Differentiation. (In press). (2004)
[文献書誌] Watanabe K, Okano H, et al.: "Comparison between fetal spinal cord-and forebrain-derived neural stem/progenitor cells as a source of transplantation for spinal cord injury."J Neurosci Res. (In press). (2004)
[文献書誌] Ohba H, Okano H, et al.: "Sox21 is a repressor of neuronal differentiation and is antagonized by YB-1."Neurosci Lett. (In press). (2004)
[文献書誌] Mikami Y, Okano H, et al.: "Implantation of dendritic cells in the injured adult spinal coed results in activation of the endogenous neural/progenitor cells for den novo neurogenesis and axonal regeneration, leading to functional recovery."J Neurosci Res. (In press). (2004)
[文献書誌] Ozawa Y, Okano H, et al.: "Down-regulation of STAT3 activation is required as an intrinsic factor to complete rod photoreceptor differentiation."Mol Cell Neurosci. (In press). (2004)
[文献書誌] Tokunaga A, Okano H et al.: "Mapping spatio-temporal activation of Notch signaling during neurogenesis and gliogenesis in the developing mouse brain."J Neurochem. (In press). (2004)

2003 年度 実績報告書

中枢神経系幹細胞の分化機構の解析

研究代表者

岡野 栄之 慶應義塾大学, 医学部, 教授 (60160694)

研究成果

[文献書誌] Tonchev AB, Okano, H, et al.: "Proliferation of neural and neuronal progenitors after global brain ischemia in young adult macaque monkeys."Mol Cell Neurosci. 23. 292-301 (2003)

[文献書誌] Kayahara T, Okano H, et al.: "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)

[文献書誌] Sasaki T, Okano H, et al.: "Implication of Cyclooxygenase-2 on Enhanced Proliferation of Neural Progenitor Cells in the Adult Mouse Hippocampus After Ischemia."J Neurosci Res. 72. 461-471 (2003)

[文献書誌] Nakamura Y, Okano H, et al.: "Expression of tubulin beta II in neural stem/progenitor cells and radial fibers during human fetal brain development"Lab Invest. 83. 479-489 (2003)

[文献書誌] Yuasa Y, Okano H, et al.: "Drosophila homeodomain protein REPO controls glial differentiation by cooperating with ETS and BTB transcription factors."Dovelopment. 130. 2419-2428 (2003)

[文献書誌] Uchida K, Okano H, et al.: "Grafted swine neuroepithelial stem cells can form myelinated axons and both efferent and afferent synapses with xenogeneic rat neurons."J Neurosci Res. 72. 661-669 (2003)

[文献書誌] Kokuzawa J, Okano H, et al.: "Hepatocyte growth factor promotes proliferation and neural differentiation of neural stem cells from mouse embryos."Mol Cell Neurosci. 24. 190-197 (2003)

[文献書誌] Ishizuya-Oka A, Okano H, et al.: "Thyroid hormone-upregulated expression of Musashi-1 is specific for progenitor cells of the adult epithelium during amphibian gastrointestinal remodeling."J Cell Sci. 116. 3157-3164 (2003)

[文献書誌] Kanuka H, Okano H, et al.: "Cytosol-endoplasmic reticulum interplay by Sec61alpha translocon in polyglutamine-mediated neurotoxicity in Drosophila."Proc Natl Acad Sai USA. 100. 11723-11728 (2003)

[文献書誌] Baker H, Okano H, et al.: "Cortical and striatal expression of tyrosine hydroxylase mRNA in neonatal and adult mice."Cell Mol Neurobiol. 23. 503-518 (2003)

[文献書誌] Tamaki T, Okano H, et al.: "Growth and differentiation potential of main- and side-population cells derived from murine skeletal muscle."Exp Cell Res. 291. 83-90 (2003)

[文献書誌] Yoshida T, Okano, H: "Distinct expression patterns of splicing isoforms of mNumb in the endocrine lineage of developing pancreas."Differentiation. 71. 486-495 (2003)

[文献書誌] Hu QD, Okano H, et al.: "F3/Contactin acts as a functional ligand for Notch during oligodendrocyte maturation."Cell. 535. 131-135 (2003)

[文献書誌] Matsuzaki Y, Kinjo K, Mulligan RC, Okano H et al.: "Drosophila homeodomain protein REPO controls glial differentiation by cooperating with ETS and BTB transcription factors."Development. 130. 2419-2428 (2003)

[文献書誌] Uchida K, Okano H, et al.: "Grafted swine neuroepithelial stem cells can form myelinated axons and both efferent and afferent synapses with xenogeneic rat neurons."J Neurosci Res. 72. 661-669 (2003)

[文献書誌] Okada S, Okano H, et al.: "Implication of Cyclooxygenase-2 on Enhanced Proliferation of Neural Progenitor Cells in the Adult Mouse Hippocampus After Ischemia."J Neurosci Res. (In press). (2004)

[文献書誌] Yamashima T, Okano H et al.: "Vascular adventitia generates neuronal progenitors in monkey hippocampus after ischemia."Hippocampus. (In press). (2004)

[文献書誌] Ieda M, Okano H, et al.: "Distinct expression patterns of splicing isoforms of mNumb in the endocrine lineage of developing pancreas."Differentiation. (In press). (2004)

[文献書誌] Watanabe K, Okano H, et al.: "Comparison between fetal spinal cord-and forebrain-derived neural stem/progenitor cells as a source of transplantation for spinal cord injury."J Neurosci Res. (In press). (2004)

[文献書誌] Ohba H, Okano H, et al.: "Sox21 is a repressor of neuronal differentiation and is antagonized by YB-1."Neurosci Lett. (In press). (2004)

[文献書誌] Mikami Y, Okano H, et al.: "Implantation of dendritic cells in the injured adult spinal coed results in activation of the endogenous neural/progenitor cells for den novo neurogenesis and axonal regeneration, leading to functional recovery."J Neurosci Res. (In press). (2004)

[文献書誌] Ozawa Y, Okano H, et al.: "Down-regulation of STAT3 activation is required as an intrinsic factor to complete rod photoreceptor differentiation."Mol Cell Neurosci. (In press). (2004)

[文献書誌] Tokunaga A, Okano H et al.: "Mapping spatio-temporal activation of Notch signaling during neurogenesis and gliogenesis in the developing mouse brain."J Neurochem. (In press). (2004)

2003 年度実績報告書

岡野栄之慶應義塾大学, 医学部, 教授 (60160694)