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，触发血压控制系统的酶主要是由肾脏中的近后细胞产生的。最近，据报道，很少的肾素是由某个地方的内脏或组织，肾上腺，脑，生殖器官和外周血血管产生的。聚合酶链反应（PCR）方法是一种有用的工具，用于检测分子生物学领域中非常小的DNA和RNA样品。这项研究的目的是使用原位转录酶（RT）-PCR方法检测和鉴定出肾外组织中表达肾素的细胞，并阐明使用杂交组织化学，cantatration和Immunoelecopron Microscopolical Microscopolical技术来阐明凝结腺肾素的功能。在前者中，修改了探针，消化，RT和PCR反应的条件，包括溶液的浓度和几个反应时间。在结果中，检测到弱反应对大鼠的淋巴结。他们被限制在毛细血管后静脉里。在后者中，可以清楚地表明，凝结腺肾素是通过外分泌方式分泌到精确管腔的，其表达受睾丸激素的调节。这些发现表明，原位RT-PCR是检测新的局部肾素 - 血管紧张素系统的有用工具，尽管该方法即使现在也在前进，并且凝结的腺肾素可能通过血管紧张素原合成对女性生殖器官作用。