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妊娠大鼠子宫中胆钙蛋白信使核糖核酸(9000Mr CaBP)的分析及原位检测

Analysis and in situ detection of cholecalcin messenger RNA (9000 Mr CaBP) in the uterus of the pregnant rat.

作者信息

Warembourg M, Perret C, Thomasset M

出版信息

Cell Tissue Res. 1987 Jan;247(1):51-7. doi: 10.1007/BF00216546.

Abstract

The molecular cloning of a cDNA fragment synthesised from rat duodenal mRNA coding for cholecalcin (calbindin), a 9000 Mr vitamin D-induced calcium-binding protein (CaBP), has been previously described. DNA/RNA hybridisation assays have been used to examine CaBP mRNA production in the uterine horns and duodena of pregnant (21 day) rats using the cloned CaBP cDNA. Northern hybridisation studies showed that the 32P cDNA sequence hybridised to a single 500-600 nucleotide species in both the uterus and the duodenum, thus demonstrating identical CaBP mRNA processing in both tissues. Dot blot hybridisation studies showed that the CaBP mRNA concentration was greatest in the duodenum while that of the uterine horns was about 10% of the duodenal level. The observed differences in CaBP mRNA levels correlate well with the in vivo CaBP concentrations. In situ hybridisation histochemistry using 3H cDNA revealed that CaBP mRNA visualised by silver grains was found in all the parts of the endometrium and the myometrium. However, CaBP mRNA was more concentrated in the outer and inner muscular fibres and in the luminal cells of the endometrium than in the stroma cells. These results demonstrate that the CaBP gene is expressed in specific cells of the rat uterus.

摘要

先前已描述了从大鼠十二指肠mRNA合成的编码胆钙化蛋白(钙结合蛋白)的cDNA片段的分子克隆,胆钙化蛋白是一种分子量为9000的维生素D诱导的钙结合蛋白(CaBP)。已使用DNA/RNA杂交试验,利用克隆的CaBP cDNA检测怀孕(21天)大鼠子宫角和十二指肠中CaBP mRNA的产生。Northern杂交研究表明,32P cDNA序列与子宫和十二指肠中的单一500 - 600个核苷酸的物种杂交,从而证明了两种组织中CaBP mRNA加工过程相同。斑点印迹杂交研究表明,CaBP mRNA浓度在十二指肠中最高,而子宫角中的浓度约为十二指肠水平的10%。观察到的CaBP mRNA水平差异与体内CaBP浓度密切相关。使用3H cDNA的原位杂交组织化学显示,通过银颗粒可视化的CaBP mRNA存在于子宫内膜和肌层的所有部分。然而,CaBP mRNA在子宫内膜的外层和内层肌纤维以及腔面细胞中比在基质细胞中更集中。这些结果表明CaBP基因在大鼠子宫的特定细胞中表达。

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