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In situ polymerase chain reaction-hybridohistochemistry for renin-producing cells.

肾素生成细胞的原位聚合酶链反应-杂交组织化学。

基本信息

批准号：
06660367
负责人：
KON Yasuhiro
金额：
$ 1.34万
依托单位：
HOKKAIDO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06660367/
关键词：
Coagulating galnd Exocrine Immunoelectron microscopy In situ hybridization Mouse Polymerase chain reaction Renin Reverse transcription RT-PCR

项目摘要

It is well known that renin, trigger enzyme of blood pressure control system is mainly produced from juxtaglomerular cells in the kidney. Recently, it has been reported that very small volume of renin is produced by somewhere extrarenal organs or tissues, adrenal gland, brain, reproductive organs and peripheral blood vessels. Polymerase chain reaction (PCR) method is a useful tool for detection of very small volume of DNA and RNA samples in molecular biological field. The aim of this study is to detect and identify renin-expressing cells in the extrarenal tissues using in situ riverse transcriptase (RT)-PCR method to histological sections, and to clarify the function of coagulating gland renin using hybridohistochemistry, castration and immunoelectron microscopical techniques. In the former, the conditions of probes, digestion, RT and PCR reactions were modified including the concentrations of solutions and several reaction times. In the result, weak reactions were detected to the lymph nodes of the rat. They were restricted to the post capillary venules. In the latter, it was cleared that coagulating gland renin was secreted to seminal lumen by exocrine manner and its expression was regulated by testosterone. These findings are showing that in situ RT-PCR is a useful tool for detection of new local renin-angiotensin system, although the method is advancing even now, and that the coagulating gland renin may effect to female reproductive organs via angiotensinogen synthesis.

众所周知，肾素是血压控制系统的触发酶，主要由肾脏肾小球旁细胞产生。近年来，有报道称肾素是由肾外器官或组织、肾上腺、脑、生殖器官和外周血管产生的。聚合酶链反应（Polymerase chain reaction， PCR）是分子生物学领域中检测小体积DNA和RNA样品的一种有效方法。本研究的目的是利用原位逆转录酶(RT)-PCR方法检测和鉴定肾素在肾外组织中的表达细胞，并利用杂交组织化学、阉割和免疫电镜技术阐明凝腺肾素的功能。前者修改了探针、酶切、RT和PCR反应的条件，包括溶液浓度和若干反应时间。结果，在大鼠淋巴结检测到弱反应。它们局限于毛细血管后小静脉。结果表明，凝腺肾素是通过外分泌方式分泌到精管内的，其表达受睾酮的调控。这些发现表明原位RT-PCR是检测局部肾素-血管紧张素新系统的有效工具，尽管该方法目前仍在发展中，并且凝固腺肾素可能通过血管紧张素原合成对女性生殖器官产生影响。