| Project/Area Number |
06557006
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 試験 |
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| Research Field |
Environmental physiology (including Physical medicine and Nutritional physiology)
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| Research Institution | KYUSHU UNIVERSITY |
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Principal Investigator |
HORI Tetsuro Kyushu University, Faculty of Medicine Professor, 医学部, 教授 (00022814)
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| Co-Investigator(Kenkyū-buntansha) |
TAKE Sachiko Kyushu University, Faculty of Medicine Assistant Professor, 医学部, 助手 (80253425)
TAKAKI Atsushi Kyushu University, Faculty of Medicine Assistant Professor, 医学部, 助手 (30243934)
KATAFUCHI Toshihiko Kyushu University, Faculty of Medicine Assistant Professor, 医学部, 講師 (80177401)
AOU Shuji Kyushu University, Faculty of Medicine Associate Professor, 医学部, 助教授 (40150908)
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| Project Period (FY) |
1994 – 1996
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| Keywords | non-RI in situ hybridization / antisense oligodeoxynucleotide / Morris water maze / hippocampus / c-kit mutant rat / LTP / c-fos / heat exposure |
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| Research Abstract |
(1) We estabalished a new and reproducible in situ hybridization technique using non-RI antisense DNA probes designed by aymmetric polymerase chain reaction and thymine dimer method. Using this technique, we revealed that high amount of IL-1betamRNA was expressed in rat splenic macrophages in responses to systemic injection of lipopolysaccharide.
(2) We synthesized an 18-mer oligodeoxynucleotide complementary to the translation initiation site of calcineurin AalphamRNA to inhibit the expression of calcineurin Aalpha and analyzed its effect on the spatial learning and memory. Injections of the antisense oligo-DNA into the bilateral hippocampus in rats resulted in a slight reduction of time required for platform finding in Morris water maze compared with animals injected with saline, although the difference was not statistically significant.
(3) Homozygous mutant rats with deficits in four aminoacids located near the tyrosine autophosphorylation site of c-kit tyrosine kinase receptor showe
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d an impaired spatial learning when tested in the Morris water maze, whereas activity and motor abilities appeared normal. In the next experiment, extracellular field potentials were recorded in areas CA1 and CA3 of the hippocampal slice preparation following stimulation of the Schaffer-commissural pathway and mossy fiber layr in the dentate gyrus, respectively. Presynaptically mediated forms of synaptic potentiation, paired-pulse facilitation of the c-kit mutant rats significantly reduced both in CA1 and CA3 region compared with+/+control rats. In addition, the mutant rats are also deficient in long-term potentiation after tetanic stimulation in CA3 region. These observations suggested a role for kit signialing in the adult central nervous system.
(4) We synthesized an 15-mer oligodeoxynucleotide complementary to the translation initiation site of c-fos mRNA to analyze roles of Fos protein in thermoregulation. When the antisense oligo-DNA was injected into the bilataral medial proptic nucleus (MPO) in rats, rectal temperatures during heat exposure were significantly lower than those in control rats injected with sense oligo-DNA.The dose of the antisense oligo-DNA was sufficient to block the expression of Fos protein in the MPO. Less
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