2005 Fiscal Year Final Research Report Summary
Regulation of molecular mechanism in growth and differentiation of dopaminergic neural progenitors and application for treatment of Parkinson's disease.
| Project/Area Number |
15590890
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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 |
Neurology
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| Research Institution | National University Corporation Tottori University |
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Principal Investigator |
TAKESHIMA Takao Tottori University, Faculty of Medicine, Associate Professor, 医学部, 助教授 (20206973)
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| Co-Investigator(Kenkyū-buntansha) |
NAKASO Kazuhiro Tottori University, University Hospital, Research Associate, 医学部附属病院, 助手 (30379648)
IMAMURA Keiko Tottori University, University Hospital, Senior Resident, 医学部附属病院, 医員 (90379652)
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| Project Period (FY) |
2003 – 2005
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| Keywords | neural progenitors / dopaminergic neuron / D-β hydroxybutyrate / PI3K-Nrf2 / THprom-GFP / rotenone |
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| Research Abstract |
We expanded mesencephalic neural progenitors from embryonic day 12 and 14 rat brain as dispersed non-attach culture. We collected RNA in the various growth/differentiation conditions and analyzed regional markers, growth-related signals and marker signals for the dopaminergic neurons. We analyzed markers such as nestin, MAP-2, TH, GABA, A2B5, vimentin, GFAP by fluorescence immunostainigs. As well as the induction of cell differentiation by fibronectin, forskolin, dopamine, and BDNF, we analyzed the effect of retinoic acid, D-β hydroxybutyrate (DBHB), and dopamine agonists. We succeeded in visualization of the TH manifestation by using a TH-gene- promoter -GFP vector in PC12 cells. We introduced the vector into the dopaminergic neural progenitors by lipofection. The induction efficiency in primary cultured progenitors was lower than in the cell line. We found that overexpression of GAPDH participated when proteasome inhibition caused degeneration of dopaminergic neurons in culture. We clarified that overexpression of GAPDH contributed to the formation of the Lewy body which is the hallmark pathology of PD brain. We also showed the participation of a change of transcription activity of p62/A170/ZIP. In the model of differentiated SH-SY5Y cell, DBHB protected the neuronal cells against the mitochondrial toxin rotenone and 3-nitropropionic acid. Furthermore, we found that deprenyl contributed to survival of the dopamine nervous system through a signal of PI3K-Nrf2.
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