1990 Fiscal Year Final Research Report Summary
Transfection of Tyrosine Hydroxylase cDNA into Cultured Cells : application for Intracerebral Grafting.
| Project/Area Number |
01480362
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Cerebral neurosurgery
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| Research Institution | National Institute of Neuroscience |
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Principal Investigator |
KOHSAKA S. Natl. Inst. Neurosci., Head, 神経研究所・代謝研究部, 部長 (50112686)
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| Co-Investigator(Kenkyū-buntansha) |
TAKEI N. Natl. Inst. Neurosci., Res. Scientist, 神経センター神経研究所・代謝研究部, 研究員 (70221372)
NAKAJIMA K. Natl. Inst. Neurosci., Res. Scientist, 神経センター神経研究所・代謝研究部, 室長 (50175494)
UCHIDA K. Natl. Inst. Neurosci., Postdoctoral fellc, 神経センター神経研究所・代謝研究部, 流動研究員 (00176687)
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| Project Period (FY) |
1989 – 1990
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| Keywords | Tyrosine hydroxylase / cDNA / transfection / L-DOPA / Transplantation / Parkinson's disease / Cultured cell / Dopamine |
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| Research Abstract |
Human Tyrosine Hydroxylase (TH) cDNA was transfected into non-neuronal cell lines (C6 glioma cell and NRK fibroblast) and feasibility of these cells as a donor material for intracerebral grafting was examined. Several transfectants that produce TH molecules were selected. In these cells, high TH activity was detected in a complete assay system in vitro. C6 cells were able to produce and secrete a large amount of L-DOPA. In contrast, NRK cells produced and secreted L-DOPA only when exogenous tetrahydrobiopterin (BH_4) was supplemented.
We investigated whether these genetically manipulated cells also express TH molecules and are able to produce L-DOPA when they are transplanted into the striatum of 6-OHDA-treated rat. All implanted C6 or NRK cells were stained by TH antiserum. Release of L-DOPA from implanted C6 cells was observed, whereas measurable release of L-DOPA from NRK cells was not found. However, intrastriatal NRK cells were able to secrete L-DOPA when BH_4 was added to the perfusates. Moreover, functional recovery was achieved by the implantation of NRK cells when BH_4 was supplemented exogenously.
These results may indicate that TH cDNA transfected non-neuronal cells can be used as donor materials for intracerebral grafting in Parkinson's model rats.
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