2001 Fiscal Year Annual Research Report

ニューロン・グリアの分化決定とニューロンの個性獲得にいたる制御機構の解析

Research Project

Project/Area Number	12210013
Research Institution	Keio University
Principal Investigator	岡野栄之慶應義塾大学, 医学部, 教授 (60160694)
Keywords	神経幹細胞 / 自己複製 / 分化調節機構 / Musashi / 水頭症 / neurosphere / アンチセンス / 生存維持
Research Abstract	本研究では、神経幹細胞の自己複製と分化調節機構に関する解析を行っていおり、本年度は、神経幹細胞に強く発現するRNA結合性蛋白質Musashiファミリーの機能解析を行った。msi1遺伝子欠損マウスのホモ接合体の出生前の胎児の剖検、組織学的観察からはmsi1遺伝子欠損マウスにおける顕著な異常は観察されなかった。神経幹細胞の増殖や分化が活発で、Msi1の発現が最も高い胎生期の脳室周囲部のventricular zone(VZ)や脳室下帯のsubventricular zone(SVZ)においても組織構築の異常を認めることはできなかった。しかし出生後約一週齢でホモ接合体では頭部の膨大が始まり2-3週齢までにドーム型の頭部を特徴とする水頭症を高頻度で(ホモ接合体の約70-80%の個体)発症した。msi1遺伝子欠損マウスは高頻度で出生後水頭症を発症し、重篤な個体は約1-2ヶ月で死亡した。水頭症を発症した成体個体を組織学的に解析した結果、側脳室の対称性の激しい拡大、大脳皮質の薄化、白質、灰白質の浮腫を伴う実質の欠損、中隔の低形成が観察された。また頭蓋内圧の亢進によると考えられる嗅球、小脳、海馬の吻側、尾側方向への圧迫変性も観察された。msi1遺伝子欠損マウスの初代培養系を用いることにより、神経幹細胞の性質を詳細に検討した。まずneurosphere法を用いて神経幹細胞の数および自己複製能を検討した。神経系の幹細胞は増殖因子存在下で浮遊培養することにより、neurosphereと呼ばれる球状の細胞塊を形成することが知られている。一定数(1x10^5細胞)の胎生14.5日終脳細胞に由来するneurosphereの数は野生型およびmsi1遺伝子欠損マウスで差はみられず、約10回の継代によっても再びneurosphereを形成することから、msi1遺伝子単独欠損によって神経幹細胞の数と自己複製能は変化しないことが明らかとなった。msi1およびmsi2遺伝子は重複した発現パターンを示しており、これらふたつの遺伝子が協同して神経幹細胞の性質を維持している可能性も考えられたため、msi1遺伝子欠損マウスのneurosphere assayにおいてmsi2遺伝子の機能をアンチセンス(antisense peptide nucleic acids ; asPNA)で抑制することにより、msi1、msi2両遺伝子を抑制した場合の神経幹細胞の性質を検討した。細胞は添加したmsi2のasPNAの濃度に依存してneurosphereの形成能を低下させることが明らかとなった。これに対して同腹の野生型由来の神経幹細胞はmsi2のasPNAの濃度には関わらず一定のneurosphereを形成していた。よってmsi1とmsi2は協調して神経幹細胞の増殖あるいは生存維持に必須な役割を担っていると考えられた。

Research Products

(25 results)

All Other

All Publications (25 results)

[Publications] Toda M, Iizuka Y, Yu W-J, Ikeda E, Yoshida K, Imai T, Kawase T, Kawakami Y, Okano H, Uyemura K: "Expression of the neural RNA-binding protein Musashi1 in human gliomas"Glia. 34. 1-7 (2001)
[Publications] Mori H, Sakakibara S, Imai T, Nakamura Y, Iijima T, Suzuki A, Yuasa Y, Takeda M, Okano H: "Expression of mouse igf2 mRNA-binding protein 3 and its implication in the developing central nervous system"J Neurosci Res. 64. 132-143 (2001)
[Publications] Murata T, Nagaso H, Watanabe S, Okano H, Yokoyama K: "The hiiragi gene encodes a poly(A) polymerase, which controls the formation of the wing margin in Drosophila melanogaster"Dev Biol. 233. 137-147 (2001)
[Publications] Okabe M, Imai T, Kurusu M, Hiromi Y, Okano H: "Translational repression determines a neuronal potential in Drosophila asymmetric cell division"Nature. 41. 94-98 (2001)
[Publications] Sawamoto K, Yamamoto A, Kawaguchi A, Yamaguchi M, Mori K, Goldman SA, Okano H: "Visualization and direct isolation of neuronal progenitor cells by dual-color flow cytometric detection of fluorescent proteins"J Neurosci Res. 65. 220-227 (2001)
[Publications] Sawamoto K, Nakao N, Kakishita K, Ogawa Y, Toyama Y, Yamamoto A, Yamaguchi M, Mori K, Goldman SA, Itakura T, Okano H: "Generation of dopaminergic neurons in the adult brain from mesencephalic precursor cells labeled with a nestin-GFP transgene"J Neurosci. 21. 3895-3903 (2001)
[Publications] Imai T, Tokunaga A, Yoshida T, Hashimoto M, Weinmaster G, Mikoshiba K, Nakafuku M, Okano H: "The neural RNA-binding protein Musashi1 translationally regulaties the m-numb gene expression by interacting with its mRNA"Mol Cell Biol. 21. 3888-3900 (2001)
[Publications] Sawamoto K, Nakao N, Kobayashi K, Matsushita N, Takahashi H, Kakishita K, Yamamoto A, Yoshizaki T, Terashima T, Murakami F, Itakura T, Okano H: "Visualization and direct isolation of midbrain dopaminergic neurons expressing GFP"Proc Natl Acad Sci USA. 98. 6423-6423 (2001)
[Publications] Uchida M, Hanai S, Uematsu N, Sawamoto K, Okano H, Miwa M, Uchida K: "Genetic and functional analysis of PARP, a DNA strand break-binding enzyme"Mutat Res. 47. 89-96 (2001)
[Publications] Ishida Y, Nakamoto H, Imai H, Suzuki S, Okano H, Suzuki H: "Heme oxygenase-1 regulates vascular tone of the peritoneum undergoing peritoneal dialysis"Advances in Peritoneal Dialysis. 17. 15-19 (2001)
[Publications] Li R-Y, Baba S, Kosugi I, Arai Y, Kawasaki H, Shinmura Y, Sakakibara S, Okano H, Tsusui Y: "Activation of murine cytomegalovirus immediate-early promoter in cerebral ventricular zone and glial progenitor cells in transgenic mice"Glia. 35. 41-52 (2001)
[Publications] Yagita Y, Kitagawa K, Otsuki T, Kuwabara K, Mabuchi T, Miyata T, Okano H, Hori M, Matsumoto M: "Proliferation of neuronal progenitor cells and increased neurogenesis in the ischemic adult rat hippocampus"Stroke. 32. 1890-1896 (2001)
[Publications] Miyata T, Kawaguchi A, Okano H, Ogawa M: "Asymmetric inheritance of radial glial fibers to neurons during corticogensis in mice"Neuron. 31. 727-741 (2001)
[Publications] Kohyama J, Abe H, Shimazaki T, Koizumi A, Nakashima K, Taga T, Okano H, Hata J, Umezawa A: "Brain from Bone : Efficient "meta-differentiation" of marrow stromal-derived mature osteoblasts to neurons with Noggin or a demethylating agant"Differentiation. 68. 235-244 (2001)
[Publications] Keyoung H.M, Roy N.S, Benraiss A, Louissaint A Jr, Suzuki A, Hashimoto K, Rashbaum WK, Okano H, Goldman SA: "Prospective identification, selection and extraction of two distinct pools of neural stem cells from fetal human brain"Nature Biotech. 19. 843-850 (2001)
[Publications] Kanemura Y, Mori K, Sakakibara S, Ohnishi T, Fujikawa H, Hayashi H, Matsumoto T, Imai T, Ohnishi T, Fushiki S, Nakamura Y, Yamasaki M, Okano H, Arita N: "Musashi1, an evolutionarily conserved neural RNA-binding protein is a versatile marker of human glioma cells in determining their cellluar origin, malignancy and proliferative activity"Differentiation. 68. 141-152 (2001)
[Publications] Sakakibara S, Nakamura Y, Satoh H, Okano H: "RNA-binding protein Musashi2 : developmentally regulated expression in neural precursor cells and subpopulation of neurons in mammalian CNS"J Neurosci. 21. 8097-8107 (2001)
[Publications] Yamamoto N, Matsunaga E, Yamamoto S, Kobayashi D, Torii M, Kawaichi M, Kishi N, Matsuno K, Okano H, Nakafuku M: "Deltex as a nuclear signal transducer downstream of the Notch receptor"J Biol Chem. 276. 45031-45940 (2001)
[Publications] Saunders PTK, Maguire SM, Macpherson S, Fenelon MC, Sakakibara S, Okano H: "The RNA binding protein Musashi1(Msi1) is expressed in the cytoplasm and nucleus of Sertoli cells in the rat testis from fetal life to adulthood"Biology of Reproduction. 66. 500-507 (2002)
[Publications] Uchida M, Hanai S, Uematsu N, Sawamoto K, Okano H, Miwa M, Uchida K: "Overexpression of poly(ADP-ribose) polymerase disrupts organization of cytoskeletal F-actin and tissue polarity in Drosophila"J Biol Chem. 277. 6696-6702 (2002)
[Publications] Matsuno K, Ito M, Hori K, Miyashita F, Suzuki S, Kishi N, Artravanis-Tsakonas S, Okano H: "Involvement of a proline-rich motif and a RING-H2 finger in a function of Deltex as a regulator of Notch signaling"Development. 129. 1049-1059 (2002)
[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. (in press). (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. (in press). (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. (in press). 141-152 (2002)
[Publications] Okano H, Sakakibara S, Sawamoto K, Nakamura Y, Kaneko Y, Akamatsu W, Tokunaga A, Imai T, Miyata T, Shimazaki T: "5th International Symposium on Tissue Engineering for Therapeutic Use"Elsevier science B.V.. 226 (2001)

2001 Fiscal Year Annual Research Report

ニューロン・グリアの分化決定とニューロンの個性獲得にいたる制御機構の解析

Principal Investigator

岡野 栄之 慶應義塾大学, 医学部, 教授 (60160694)

Research Products

[Publications] Toda M, Iizuka Y, Yu W-J, Ikeda E, Yoshida K, Imai T, Kawase T, Kawakami Y, Okano H, Uyemura K: "Expression of the neural RNA-binding protein Musashi1 in human gliomas"Glia. 34. 1-7 (2001)

[Publications] Mori H, Sakakibara S, Imai T, Nakamura Y, Iijima T, Suzuki A, Yuasa Y, Takeda M, Okano H: "Expression of mouse igf2 mRNA-binding protein 3 and its implication in the developing central nervous system"J Neurosci Res. 64. 132-143 (2001)

[Publications] Murata T, Nagaso H, Watanabe S, Okano H, Yokoyama K: "The hiiragi gene encodes a poly(A) polymerase, which controls the formation of the wing margin in Drosophila melanogaster"Dev Biol. 233. 137-147 (2001)

[Publications] Okabe M, Imai T, Kurusu M, Hiromi Y, Okano H: "Translational repression determines a neuronal potential in Drosophila asymmetric cell division"Nature. 41. 94-98 (2001)

[Publications] Sawamoto K, Yamamoto A, Kawaguchi A, Yamaguchi M, Mori K, Goldman SA, Okano H: "Visualization and direct isolation of neuronal progenitor cells by dual-color flow cytometric detection of fluorescent proteins"J Neurosci Res. 65. 220-227 (2001)

[Publications] Sawamoto K, Nakao N, Kakishita K, Ogawa Y, Toyama Y, Yamamoto A, Yamaguchi M, Mori K, Goldman SA, Itakura T, Okano H: "Generation of dopaminergic neurons in the adult brain from mesencephalic precursor cells labeled with a nestin-GFP transgene"J Neurosci. 21. 3895-3903 (2001)

[Publications] Imai T, Tokunaga A, Yoshida T, Hashimoto M, Weinmaster G, Mikoshiba K, Nakafuku M, Okano H: "The neural RNA-binding protein Musashi1 translationally regulaties the m-numb gene expression by interacting with its mRNA"Mol Cell Biol. 21. 3888-3900 (2001)

[Publications] Sawamoto K, Nakao N, Kobayashi K, Matsushita N, Takahashi H, Kakishita K, Yamamoto A, Yoshizaki T, Terashima T, Murakami F, Itakura T, Okano H: "Visualization and direct isolation of midbrain dopaminergic neurons expressing GFP"Proc Natl Acad Sci USA. 98. 6423-6423 (2001)

[Publications] Uchida M, Hanai S, Uematsu N, Sawamoto K, Okano H, Miwa M, Uchida K: "Genetic and functional analysis of PARP, a DNA strand break-binding enzyme"Mutat Res. 47. 89-96 (2001)

[Publications] Ishida Y, Nakamoto H, Imai H, Suzuki S, Okano H, Suzuki H: "Heme oxygenase-1 regulates vascular tone of the peritoneum undergoing peritoneal dialysis"Advances in Peritoneal Dialysis. 17. 15-19 (2001)

[Publications] Li R-Y, Baba S, Kosugi I, Arai Y, Kawasaki H, Shinmura Y, Sakakibara S, Okano H, Tsusui Y: "Activation of murine cytomegalovirus immediate-early promoter in cerebral ventricular zone and glial progenitor cells in transgenic mice"Glia. 35. 41-52 (2001)

[Publications] Yagita Y, Kitagawa K, Otsuki T, Kuwabara K, Mabuchi T, Miyata T, Okano H, Hori M, Matsumoto M: "Proliferation of neuronal progenitor cells and increased neurogenesis in the ischemic adult rat hippocampus"Stroke. 32. 1890-1896 (2001)

[Publications] Miyata T, Kawaguchi A, Okano H, Ogawa M: "Asymmetric inheritance of radial glial fibers to neurons during corticogensis in mice"Neuron. 31. 727-741 (2001)

[Publications] Kohyama J, Abe H, Shimazaki T, Koizumi A, Nakashima K, Taga T, Okano H, Hata J, Umezawa A: "Brain from Bone : Efficient "meta-differentiation" of marrow stromal-derived mature osteoblasts to neurons with Noggin or a demethylating agant"Differentiation. 68. 235-244 (2001)

[Publications] Keyoung H.M, Roy N.S, Benraiss A, Louissaint A Jr, Suzuki A, Hashimoto K, Rashbaum WK, Okano H, Goldman SA: "Prospective identification, selection and extraction of two distinct pools of neural stem cells from fetal human brain"Nature Biotech. 19. 843-850 (2001)

[Publications] Sakakibara S, Nakamura Y, Satoh H, Okano H: "RNA-binding protein Musashi2 : developmentally regulated expression in neural precursor cells and subpopulation of neurons in mammalian CNS"J Neurosci. 21. 8097-8107 (2001)

[Publications] Yamamoto N, Matsunaga E, Yamamoto S, Kobayashi D, Torii M, Kawaichi M, Kishi N, Matsuno K, Okano H, Nakafuku M: "Deltex as a nuclear signal transducer downstream of the Notch receptor"J Biol Chem. 276. 45031-45940 (2001)

[Publications] Saunders PTK, Maguire SM, Macpherson S, Fenelon MC, Sakakibara S, Okano H: "The RNA binding protein Musashi1(Msi1) is expressed in the cytoplasm and nucleus of Sertoli cells in the rat testis from fetal life to adulthood"Biology of Reproduction. 66. 500-507 (2002)

[Publications] Uchida M, Hanai S, Uematsu N, Sawamoto K, Okano H, Miwa M, Uchida K: "Overexpression of poly(ADP-ribose) polymerase disrupts organization of cytoskeletal F-actin and tissue polarity in Drosophila"J Biol Chem. 277. 6696-6702 (2002)

[Publications] Matsuno K, Ito M, Hori K, Miyashita F, Suzuki S, Kishi N, Artravanis-Tsakonas S, Okano H: "Involvement of a proline-rich motif and a RING-H2 finger in a function of Deltex as a regulator of Notch signaling"Development. 129. 1049-1059 (2002)

[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. (in press). (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. (in press). 141-152 (2002)

[Publications] Okano H, Sakakibara S, Sawamoto K, Nakamura Y, Kaneko Y, Akamatsu W, Tokunaga A, Imai T, Miyata T, Shimazaki T: "5th International Symposium on Tissue Engineering for Therapeutic Use"Elsevier science B.V.. 226 (2001)

岡野栄之慶應義塾大学, 医学部, 教授 (60160694)