RT-PCR microlocalization of mRNA for guanylyl cyclase-coupled ANF receptor in rat kidney.
RT-PCR microlocalization of mRNA for guanylyl cyclase-coupled ANF receptor in rat kidney.
复制标题
大鼠肾脏中鸟苷酸环化酶偶联 ANF 受体 mRNA 的 RT-PCR 微定位。
DOI：
10.1152/ajprenal.1991.261.6.f1080
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发表时间：
1991
期刊：
影响因子：
--
通讯作者：
A. García
中科院分区：
文献类型：
--
作者：
Y. Terada;T. Moriyama;B. Martin;M. Knepper;A. García
在大鼠肾脏中进行了冠酰环化酶偶联心房利钠因子（ANF）受体mRNA编码微定位。我们在单个微解剖肾小管段、肾小球和直血管束中使用了逆转录和聚合酶链反应（RT-PCR）的组合。利用特异32p标记探针探测的Southern blots自放射图密度测定法对扩增的cDNA进行相对定量。在肾小管段中，终末髓内集管（IMCD）信号最大。在初始IMCD和内髓的Henle节袢中发现稍小的信号。在以下部分（由高到低）也可见到易检测到的信号：皮质集管、近曲小管、髓质厚升肢、皮质厚升肢、远曲小管和髓质外集管。在肾小球和外髓质内条纹的直血管束中也检测到大信号。基于这些结果，我们得出结论：1)通过在微解剖的肾小管和血管元件中应用RT-PCR程序，在肾脏微定位编码激素受体的特异性mrna是可行的；2)guanyyl环化酶偶联ANF受体的基因沿肾元广泛表达，这提高了存在多个ANF作用位点的可能性。
Microlocalization of mRNA coding for the guanylyl cyclase-coupled atrial natriuretic factor (ANF) receptor was carried out in the rat kidney. We used a combination of reverse transcription and polymerase chain reaction (RT-PCR) in individual microdissected renal tubule segments, glomeruli, and vasa recta bundles. Relative quantitation of the resulting amplified cDNA utilized densitometry of autoradiograms from Southern blots probed with a specific 32P-labeled probe. Among renal tubule segments, the largest signal was found in the terminal inner medullary collecting duct (IMCD). Slightly smaller signals were found in the initial IMCD and in loop of Henle segments from the inner medulla. Readily detectable signals were also seen in the following segments (in descending order): cortical collecting duct, proximal convoluted tubule, medullary thick ascending limb, cortical thick ascending limb, distal convoluted tubule, and outer medullary collecting duct. Large signals were also detected in glomeruli and in vasa recta bundles from the inner stripe of the outer medulla. Based on these results, we conclude that 1) renal microlocalization of specific mRNAs coding for hormone receptors is feasible through application of the RT-PCR procedure in microdissected renal tubules and vascular elements, and 2) the gene for the guanylyl cyclase-coupled ANF receptor is broadly expressed along the nephron, raising the possibility that multiple sites of ANF action are present.