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利用原位杂交技术对大鼠脑海马体中糖皮质激素受体信使核糖核酸进行定位。

Localization of glucocorticoid receptor messenger ribonucleic acid in hippocampus of rat brain using in situ hybridization.

作者信息

Yang G, Matocha M F, Rapoport S I

机构信息

Laboratory of Neurosciences, National Institute on Aging, Bethesda, Maryland 20892.

出版信息

Mol Endocrinol. 1988 Aug;2(8):682-5. doi: 10.1210/mend-2-8-682.

Abstract

An in situ hybridization procedure was applied to quantify glucocorticoid receptor (GR) mRNAs in the hippocampus of rat brain. Hybridization was carried out using a radiolabeled antisense probe complementary to the rat liver GR gene. The specificity of the method was validated by showing: 1) a high cellular grain density in sections hybridized with an antisense but not a sense probe; 2) agreement between the experimental and theoretical temperature at which 50% of the hybrids melted, and 3) a high signal distribution of GR mRNA in the hippocampus, a region of brain known to preferentially concentrate steroid hormones. Within the hippocampus, however, subregional differences in hybridization densities were observed. Quantitative autoradiography indicated that the average neuronal silver grain number was highest in the pyramidal cell layers of CA2 and CA4 and lowest in those of CA1 and CA3. Also, there was a significant difference in the average grain number between all of the cell fields except for that between CA2 and CA4. These results show that contiguous but neuroanatomically distinct cell fields of the hippocampus express different levels of GR transcripts, and indicate that differential regulation of GR expression occurs in subpopulations of hippocampal neurons.

摘要

应用原位杂交技术对大鼠脑海马区糖皮质激素受体(GR)mRNA进行定量分析。采用与大鼠肝脏GR基因互补的放射性标记反义探针进行杂交。该方法的特异性通过以下几点得到验证:1)与反义探针而非正义探针杂交的切片中细胞颗粒密度高;2)50%杂交体解链的实验温度与理论温度一致;3)GR mRNA在海马区有高信号分布,海马区是已知优先浓缩甾体激素的脑区。然而,在海马区内,观察到杂交密度存在亚区域差异。定量放射自显影表明,平均神经元银颗粒数在CA2和CA4的锥体细胞层最高,在CA1和CA3的锥体细胞层最低。此外,除CA2和CA4之间外,所有细胞区域的平均颗粒数均存在显著差异。这些结果表明,海马区相邻但神经解剖学上不同的细胞区域表达不同水平的GR转录本,提示海马神经元亚群中存在GR表达的差异调节。

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