| Co-Investigator(Kenkyū-buntansha) |
TAKE Sachiko Kyushu University, Dept. of Physiology, Assistant Professor, 大学院・医学系研究科, 助手 (80253425)
TAKAKI Atsushi Kyushu University, Dept. of Physiology, Assistant Professor, 大学院・医学系研究科, 助手 (30243934)
KATAFUCHI Toshihiko Kyushu University, Dept. of Physiology, Assistant Professor, 大学院・医学系研究科, 講師 (80177401)
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| Research Abstract |
We have developed the methods for to analyze the expression of brain cytokines and an immediate eary gene, c-fos, during the stress response to various environmental stimuli using molecular biological techniques. We also examined the effects of microinjection of antisense oligo DNA for c-fos mRNA into the hypothalamus on the responses to the stressors. Our findings are as follows ;
(1) Cytokines including IL-1β and IFNα were expressed in the brain in response to not only inflammatory stresses but also non-inflammatory stresses such as immobilization (IMB) and heat (33℃) or cold (4℃) exposure.
(2) Quantitative analyses of the mRNA for brain cytokines and neuropeptide were successively performed by competitive RT-PCR using mimic cDNA and real-time capillary RT-PCR methods.
(3) We observed an expression of Fos protein in the medial preoptic area (MPO), paraventricular nucleus (PVN), and ventromedial nucleus (VMH) in the hypothalamus after IMB and heat or cold exposure for 2 hours.
(4) Express
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ion of c-fos mRNA equally increased in the respective hypothalamus nuclei in rats by heat exposure when measured by real-time papillary RT-PCR methods. However, cold exposure induced marked increases in the amount of c-fos mRNA in the MPO, lateral hypothalamus (LHA), and VMH.
(5) Heat exposure produced a decrease in IL-1β and an increase in IFN-αmRNA contents, while mRNAs for IL-6 and TNF-α were not affected.
(6) For protein expressed in rat MPO during heat or cold exposure works to elevate body temperature since blockage of Fos expression by antisense oligo DNA for c-fos mRNA results in the decrease in body temperature during both heat and cold exposure.
(7) The suppression of Fos protein induced by antisense oligo DNA was accompanied by the increase in the amount of c-fos mRNA in the MPO, suggesting an autoinhibitory action of Fos protein on the expression of c-fos gene. This also suggests that the suppression of Fos protein by antisense oligo DNA results from an inhibition of the translation of c-fos mRNA. Furthermore, since the expression of IFN-αmRNA by heat exposure was suppressed by antisense oligo DNA, it is suggested that one of the target genes of Fos protein is IFN-α gene. Less
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