| Project/Area Number |
15590207
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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 |
Environmental physiology (including Physical medicine and Nutritional physiology)
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| Research Institution | Shizuoka University |
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Principal Investigator |
YAMADA Junko Shizuoka University, Graduate School of Electronic Science and Technology, Research Associate, 大学院・電子科学研究科, 助手 (30334965)
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| Co-Investigator(Kenkyū-buntansha) |
FUKUDA Atsuo Hamamatsu University School of Medicine, Department of physiology, Professor, 医学部, 教授 (50254272)
INOUE Koichi Hamamatsu University School of Medicine, Department of physiology, Research Associate, 医学部, 助手 (80345818)
OKABE Akihito Hamamatsu University School of Medicine, Department of physiology, Research Associate, 医学部, 助手 (10313941)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 2004: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2003: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | development / Chrolide / Cl^-homeostasis / GABA receptor / Tributyotin / GFP / in utero electropolation / neocortex / 神経回路 / シナプス / 電気穿孔法 |
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| Research Abstract |
Measurement of [Cl^-]i and cell miglation
Using in vivo electropolation, we transfected eGFP prasmid embryonic day 14 (E14) mice. After electropolation, mice were exposed tributyltin chloride (TBTCl ; 1μl) via amniotic fluid. 3-4 days after exposure TBTCl, mice were observed their conditions.
Results were as follows ;
1.TBTCl 1M : Mice were dead with atrophy. Both side of mice, which were intact, also dead.
2.500 mM-100 mM : Mice were dead with atrophy. Both side of mice, which were intact, survived.
3.10 mM : Mice were dead but looks were normal.
4.1 mM : Mice sere survived and cell miglation was normal.
The effect of TBTCl exposure on Cl^-homeostasis.
Using gramicidin perforate patch-clamp method, we measured [Cl^-]_i TBTCl exposed cell (30 nM, 3days) and intact cell (0.1 % DMSO, 3 days). Both cell group had normal [Cl^-]_i.
The effect of TBTCl exposure on GABA receptor expression
mRNAs of TBTCl exposed cell and control cell were harvested. Using RT-PCR method, expression of GABAA receptor subunit (α1-5、β1-3、γ1-3、δ、ε、and β-actin)were analysed.
The acute effect of TBTCl on culture cell.
Using patch-clamp method and Ca2+ imaging, neurons were observed effect of TBTCl application. At low concentration of TBTCl application (30 nM), there were no effect on membrane potantial and synaptic current. It seems that toxicity at low concentration of TBTCl acts slowly.
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