| Project/Area Number |
62870100
|
|---|
| Research Category |
Grant-in-Aid for Developmental Scientific Research
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
医学一般
|
|---|
| Research Institution | Okayama University, Faculty of Pharmaceutical Sciences |
|---|
Principal Investigator |
TSUDA Masaaki Okayama Univ., Fac. of Pharmaceutical Sci., Associate Professor, 薬学部, 助教授 (80132736)
|
|---|
| 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)
|
|---|
| Project Period (FY) |
1987 – 1989
|
| Project Status |
Completed (Fiscal Year 1989)
|
| Budget Amount *help |
¥9,300,000 (Direct Cost: ¥9,300,000)
Fiscal Year 1989: ¥3,800,000 (Direct Cost: ¥3,800,000)
Fiscal Year 1988: ¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1987: ¥3,100,000 (Direct Cost: ¥3,100,000)
|
|---|
| Keywords | Retrovirus-mediated gene transfer / Cerebellum / Primary culture / Transplantation / Gene therapy / ベクター / 中枢神経系 / 移植 / ウイルス感染 / 神経疾患治療 |
|---|
| 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.
|
