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大鼠下丘脑肌动蛋白mRNA水平的昼夜变化以及35S-甲硫氨酸掺入肌动蛋白的情况。

Diurnal changes in actin mRNA levels and incorporation of 35S-methionine into actin in the rat hypothalamus.

作者信息

Iovanna J, Dusetti N, Calvo E, Cardinali D P

机构信息

Departamento de Fisiología, Facultad de Medicina, Universidad de Buenos Aires, Argentina.

出版信息

Cell Mol Neurobiol. 1990 Jun;10(2):207-16. doi: 10.1007/BF00734574.

Abstract
  1. The in vitro incorporation of 35S-methionine into actin and total soluble proteins, as well as the levels of actin mRNA, were studied in the hypothalamus and frontal cerebral cortex of adult male rats killed at six different time intervals during a 24-hr cycle. 2. The specific activity of total soluble proteins after labeled methionine incubations did not vary as a function of time of day in any of the examined brain regions. 3. The incorporation of 35S-methionine into a 43-kDa protein, corresponding to the electrophoretic mobility of actin, varied diurnally in the hypothalamus, exhibiting a maximum at 1200 hr. Such a diurnal variation was not found in frontal cerebral cortex. 4. Similar results were obtained when labeled methionine incorporation into actin was assessed in hypothalamus and cerebral cortex by an immunoprecipitation procedure. 5. An increase in actin hypothalamic mRNA levels, quantitated by dot-blot analysis, was found at 0800, 4 hr in advance to the maximum in 35S-methionine incorporation to actin. 6. The levels of actin mRNA did not vary significantly as a function of time of day in the frontal cerebral cortex.
摘要
  1. 研究了在24小时周期内,于六个不同时间点处死的成年雄性大鼠的下丘脑和额叶皮质中,35S-甲硫氨酸在体外掺入肌动蛋白和总可溶性蛋白的情况,以及肌动蛋白mRNA的水平。2. 在任何一个被检查的脑区中,经标记甲硫氨酸孵育后总可溶性蛋白的比活性均不会随一天中的时间而变化。3. 35S-甲硫氨酸掺入对应于肌动蛋白电泳迁移率的43 kDa蛋白的情况,在下丘脑中呈昼夜变化,在12:00时达到最大值。在额叶皮质中未发现这种昼夜变化。4. 当通过免疫沉淀法评估下丘脑和大脑皮质中标记甲硫氨酸掺入肌动蛋白的情况时,得到了类似的结果。5. 通过斑点印迹分析定量发现,下丘脑肌动蛋白mRNA水平在08:00时升高,比35S-甲硫氨酸掺入肌动蛋白的最大值提前4小时。6. 额叶皮质中肌动蛋白mRNA的水平不会随一天中的时间而显著变化。

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