1989 Fiscal Year Final Research Report Summary
Application of retrovirus-mediated gene transfer and cell transplantation for studying the structure and function of nervous systems at the level of gene action
| Project/Area Number |
62870100
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| Research Category |
Grant-in-Aid for Developmental Scientific Research
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| Allocation Type | Single-year Grants |
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| Research Field |
医学一般
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| Research Institution | Okayama University, Faculty of Pharmaceutical Sciences |
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Principal Investigator |
TSUDA Masaaki Okayama Univ., Fac. of Pharmaceutical Sci., Associate Professor, 薬学部, 助教授 (80132736)
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| Co-Investigator(Kenkyū-buntansha) |
ONO Katsuhiko Okayama Univ., Fac. of Medical Sci., Research associate, 医学部, 助手 (30152523)
KAWAMURA Koki Keio Univ., Fac. of Medical Sci., Professor, 医学部, 教授 (40048286)
TSUCHIYA Tomofusa Okayama Univ., Fac. of Pharmaceutical Sci., Professor, 薬学部, 教授 (80012673)
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| Project Period (FY) |
1987 – 1989
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| Keywords | Retrovirus-mediated gene transfer / Cerebellum / Primary culture / Transplantation / Gene therapy |
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| Research Abstract |
For understanding the structure and function of the central nervous systems (CNS), it is convenient to introduce foreign genes into specific areas of brain and express them. In addition, this method should be useful in the treatment of nervous disorders caused by the loss or damage of certain cells in specified areas of the CNS.
To develop a procedure for the transplantation of genetically modified brain primary cells, we transplanted cultured mouse cerebellar cells infected with recombinant retroviruses harboring chloramphenicol acetyltransferase (CAT) gene, we selected only virus-infected cells for transplantation by culturing the cells in medium containing G418 for 3 weeks. CAT was continuously expressed in the cultured cerebellar cells during 3 week incubation, but by immunoblotting analysis with anti-glial fibtillar acidic protein (GFAP) or anti-neurofilament protein (NFP) antiserum the population of cultured cerebellar cells was found to change during the incubation. Immunohistochemical analysis using anti-CAT antiserum demonstrated that the transplanted cell mass containing CAT-positive cells was detectable in the cerebellum up to 3 weeks, but 3 months after the transplantation of G418-selected cells into the cerebellar of 7-week-old mice. Recently, we have found that the plasmid DNAs injected into mouse brain through microsyringe can be incorporated and expressed by brain cells. This method would give us a very convenient method for studying the structure and function of the CNS at the level of gene regulation.
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[Publications] Tsuda, M., Ono, K., Katayama, N., Yamagata, Y., Kikuchi, K. and Tsuchiya, T.: "Neurite outgrowth from mouse neuroblastoma and cerebellar cells induced by the protein kinase inhibitor H-7" Neurosci. Lett. 105, 241-245 (1989).
Description
「研究成果報告書概要(欧文)」より
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[Publications] Katayama, N., Yamagata, Y., Hashimoto, H., Kanazawa, H., Tsuchiya, T. and Tsuda, M.: "Concanavalin A affects -tubulin mRNA expression during neuritic processes of mouse neuroblastoma N18TG2 cells in a different manner from colchicine" Biochem. Biophys. Res. Commun. (1990).
Description
「研究成果報告書概要(欧文)」より
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[Publications] Tsuda, M., Yuasa, S., Fujino, Y., Sekikawa, M., Ono, K., Tsuchiya, T. and Kawamura, K.: "Retrovirus-mediated gene transfer into mouse cerebellar primary culture and its application to the neural transplantation" Brain Res. Bull. (1990).
Description
「研究成果報告書概要(欧文)」より
