1994 Fiscal Year Final Research Report Summary
Identification and functional analysis of proteins involved in neural circuit formation
| Project/Area Number |
05680679
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Neurochemistry/Neuropharmacology
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| Research Institution | Tokyo Metropolitan University |
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Principal Investigator |
ISOBE Toshiaki Tokyo Metropolitan Univ, Associate professor, 理学部, 助教授 (70106607)
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| Co-Investigator(Kenkyū-buntansha) |
HANAOKA Kazunori Kitazato Univ.Professor, 理学部, 教授 (40189577)
ISOBE Toshiaki Tokyo Metropolitan Univ, Associate professor (70106607)
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| Project Period (FY) |
1993 – 1994
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| Keywords | Neural circuit / Cerebellar morphogenesis / Cell cycle / Signal transduction / Cerebellar protein / molecular cloning / Nuclear factor / Primary structure |
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| Research Abstract |
Morphogenesis of the rat cerebellar cortex, including differentiation of neurons and synaptogenesis, occurs at an early postnatal stage, mainly during the first two weeks after birth. To elucidate the molecular mechanism of this dynamic cellular process, we have developed a data base of rat cerebellar proteins detectable by two-dimensional electrophoresis. The data base contains two-dimensional patterns and derived information pertaining to polypeptide constituents of the developing rat cerebellum. Using this data base, we have compared the protein constituents of a series of immature and mature cerebella. A subset of polypeptides were found to be uniquely expressed in the immature cerebellum. A number of polypeptides of this class were digested by proteinases in situ or after extraction from the gel, and the generated fragments were micro-sequenced by conventional Edman degradation or mass spectrometry combined with liquid chromatography. The sequences were then compared with those in
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protein/gene sequence data bases to identify the isolated proteins. The identified proteins include phosphorylated and non-phosphorylated forms of stathmin whichi has been studied extensively in relation to the differentiation of neurons, and fatty acid-binding protein that belongs to a family of retinoic-acid binding proteins and participates in the neuronal differentiation in chick retina. Two proteins analyzed were unique in that no significant homologies were found to any other sequences in the data bases. The amino acid sequence of these proteins termed V-1 and LANP respectively, were determined by direct protein analysis and cDNA cloning. The sequence showed that the V-1 is a typical cytosolic protein of 117 amino acids and has two and a half of 33 amino acids repeats called "cell-cycle motif". LAMP contains 247 amino acids consisting of teh N-teminally located leucine-rich repeats followed by an acidic amino acids tail with nuclear transfer signal. Both the cell-cycle motif and leucine-rich repeats mediate protein-protein interation through binding to specific target molecules, and are found in numbers of proteins that function in cell fate determination in wide organisms including yeast and Drosophila. In situ histochemical analysis postnatal rat cerebella showed the transient expression of V-1 in granular layr and of LANP in Purkinje cells, suggsting the potential roles of V-1 and LANP in the signal transduction pathway that directs the cerebellar morphogenesis. Less
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Research Products
(18 results)
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[Publications] Taoka, M., Isobe, T., Okuyama, T., Yamakawa, Y., Watanabe, M., Kondo, H., Ozawa, F., Hishinuma, F., Noguchi, K., Song, S-Y.and Yamakuni, T: "Murine cerebellar neurons express a novel gene encoding a protein related to cell-cycle control and cell-fate determination proteins." J.Biol.Chem.269. 9946-9951 (1994)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Matsuoka, K., Taoka, M., Satozawa, N., Nakayama, H., Ichimura, T., Takahashi, N., Yamakuni, T., Song, S.-Y.and Isobe, T: "A nuclear factor containing the leucine-rich repeats expressed in murine cerebellar neurons." Proc.Natl.Acad.Sci.U.S.A. 91. 9670-9674 (1994)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Matsuoka, K., Seta, K., Yamakawa, Y., Okuyama, T., Shinoda, T.and Isobe, T: "The covalent structure of bovine brain calreticulin." Biochem.J. 298. 435-442 (1994)
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「研究成果報告書概要(欧文)」より
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[Publications] Shimizu, K., Kuroda, S., Yamamori, B., Matsuda, S., Kaibuchi, K., Yamauchi, T., Isobe, T., Irie, K., Matsumoto, K.and Takai, Y.: "Synergistic activation by Ras and 14-3-3 protein of a MAP kinase kinase kinase named Ras-dependent ERK kinase stimulator (REKS)." J.Biol.Chem.269. 22917-22920 (1994)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Watanabe, M., Isobe, T., Ichimura, T., Kuwano, R., Takahashi, Y.and Kondo, H: "Molecular cloning of rat cDNAs for beta and gamma subtypes of 14-3-3 protein and developmental changes in expression of their mRNAs in the nervous system." Mol.Brain Res. 17. 135-146 (1994)
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「研究成果報告書概要(欧文)」より
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[Publications] Watanabe, M., Isobe, T., Ichimura, T., Kuwano, R., Takahashi, Y.and Kondo, H.(1993): "Develomental regulation of neuronal expression for the eta subtype of the 14-3-3 protein, a putative regulatory protein for protein kinase C." Dev.Brain Res. 73. 225-235 (1993)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Watanabe, M., Isobe, T., Ichimura, T., Kuwano, R., Takahashi, Y.and Kondo, H.and Inoue, Y.: "Molecular cloning of rat cDNAs for the zeta andsigma subtypes of 14-3-3 protein and differential distributions of their mRNAs in the brain." Mol.Brain Res. 25. 113-121 (1994)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Sawai, S., Shimono, A., Wakamatsu, Y., Palmes, C., Hanaoka, K.& Kondoh, H: "Defects of embryonic organogenesis resulting from targeted disruption of the N-myc gene in the mouse." Development. 117. 1445-1455 (1993)
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「研究成果報告書概要(欧文)」より
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[Publications] Nabeshima, Y., Hanaoka, K., Hayasaka, M., Showei, Li., Esumi, Y.& Nabeshima, Y.: "Myogenin gene disruption results in perinatal lethality because of severe muscle defects." Nature. 364. 332-335 (1993)
Description
「研究成果報告書概要(欧文)」より
