| Project/Area Number |
13671556
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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 |
Anesthesiology/Resuscitation studies
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| Research Institution | HOKKAIDO UNIVERSITY |
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Principal Investigator |
MORIMOTO Yuji Hokkaido Univ., Grad. School of Med., Assistant Prof., 医学部附属病院, 講師 (00250457)
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| Project Period (FY) |
2001 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2002: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 2001: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | Cell culture ; cortical neuron / Cell death apoptosis / Hypothermia ; mild hypothermia / Barbiturate ; pentobarbital / Caspase / Reactive oxygen species / Cerebral ischemia / Glutamate ; glutamate neurotoxocity / 神経細胞培養 / グルタミン酸 |
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| Research Abstract |
It has been reported that mild hypothermia and anesthetics such as barbiturates are neuroprotecitve. However, the mechanism of this protective effect has not been well clarified. We evaluated the effect of hypothermia and pentobarbital on glutamate neurotoxicity in rat cortical neurons.
Rat embryonic cortical neurons were obtained from timed pregnant rats at 18 days gestation. They were maintained in serum-free medium (Neurobasal medium supplemented with B-27 and N-2) for 6 days. The nearly pure neuronal population was obtained by this culture system. After exposure to 50 μM glutamate for 30 min, hypothermia or pentobarbital was induced. After 24 hours, neurotoxicity was evaluated by a leakage assay of lactic dehydogenase. Neuroprotective effect was seen in hypothermia (33 and 30 ℃), but not in clinical doses of pentobarbital. However, induction of hypothermia only during exposure to glutamate did not show any protective effect. Second, production of ROS (measured by flow cytometry with a ROS-specific fluorogen, C-DCDHF-DA) and caspase-3 activation (measured by DEVD-p-nitroaniline cleavage assays) was not inhibited by hypothermia after exposure of glutamate.
Our data indicated that hypothermia is neuroprotective against exposure to glutamate by inhibiting any cascade after the exposure, but not by inhibiting direct effect of glutamate during exposure. However, we did not make clear what cascade was inhibited by hypothermia in this study.
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