Analysis of homeostatic responses to stressful environments using molecular techniques.

使用分子技术分析对应激环境的稳态反应。

基本信息

批准号：
06557006
负责人：
HORI Tetsuro
金额：
$ 11.26万
依托单位：
KYUSHU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06557006/
关键词：
non-RI in situ hybridization antisense oligodeoxynucleotide Morris water maze hippocampus c-kit mutant rat LTP c-fos heat exposure ミュータントラット paired pulse fancilitation 暑熱暴露拘束ストレスインターロイキン-6 肝臓交感神経海馬モリス水迷路 mRNA 非放射性cDNA

项目摘要

(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 … More 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

（1）我们使用该技术使用该技术的NON-RI ANTISSE DNA探针，使用非RI AntisonSe DNA探针进行了新的和可重现的原位杂交技术。脾巨噬细胞对全身性含糖的反应（2）。大鼠的双侧海马是由于盐水注射液体的较紧密的伪造发现，以索引indis水迷宫，（3）在四个氨基酸的位置位置的纯合突变体附近的纯合突变体。在下一个经验中，更多的空间学习wat迷宫是正常的。与+/+对照大鼠相比，齿状纤维中的纤维纤维在1和CA3区域中都显着介导观察表明，在成年中央神经系统中，我们将15-mer的寡脱氧核苷酸合成到C-FOS mRNA的翻译位置，以分析FOS蛋白在热疗法中的作用。在大鼠中，双腹温度的内侧内侧支撑核（MPO）显着降低了注射寡-DNA的托洛尔大鼠。反义寡-DNA的剂量令人震惊，以阻止MPO中的FOS蛋白质