2001 Fiscal Year Final Research Report Summary
Establishment of Midbrain Dopaminergic Cell Line and Its Application to Screening of Striatum Neurotrophic Factors
| Project/Area Number |
11672196
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biological pharmacy
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| Research Institution | Meijo University |
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Principal Investigator |
KANEDA Norio Meijo University, Faculty of Pharmacy, Professor, 薬学部, 教授 (00144139)
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| Co-Investigator(Kenkyū-buntansha) |
HIKITA Kiyomi Meijo University, Faculty of Pharmacy, Research Associate, 薬学部, 助手 (30257654)
MURATA Tomiyasu Meijo University, Faculty of Pharmacy, Research Associate, 薬学部, 助手 (80285189)
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| Project Period (FY) |
1999 – 2001
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| Keywords | Parkinson Disease / brain grafting / transgenic mice / immortalization / adrenal medulla / dopaminergic / neurotrophic factor |
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| Research Abstract |
For the purpose of developing neural cell line which is useful for the treatment of Parkinson disease, we planned to establish a catecholaminergic cell line using transgenic (Tg) mouse in which an oncogene, ts-mutant of SV40T antigen is expressed under the control of tyrosine hydroxylase (TH) gene promoter. These Tg mice were found to generate frequently adrenal tumor about several months after birth.
In 1999, we studied expression profiles of intrinsic TH and T antigen in these mice. Whereas T antigen was strongly expressed in the adrenal gland of adult mice, no expression was detected in the substantia nigra. At E13.5, overlapped expression of T antigen and intrinsic TH were detected in the ventral region of mesencephalon.
In 2000, genetically manipulated adrenal cells were isolated from the tumor of the Tg mouse, and the optimal culture conditions were examined. Collagen was the best substrate for the culture of these cells, and we isolated two adrenal chromaffin cell lines, one of which, named tsAM5D, was a dopamine-producing cell line. These cells expressed both TH and T antigen at a permissive temperature (33 ℃), but the latter was disappeared at non-permissive temperature (39 ℃).
In 2001, dependency of tsAM5D cells on various neurotrophic factors was examined. The cells required CNTF and FGF-2 for their growth at 33 ℃, but showed only a weak response to NGF or NT-3. The cells differentiated morphologically into neurons in the presence of CNTF and FGF-2 at 39 ℃.
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