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

众所周知，肾素是血压控制系统的触发酶，主要由肾脏的肾小球旁细胞产生。最近，有报道称肾外某些器官或组织、肾上腺、脑、生殖器官和外周血管产生了极少量的肾素。聚合酶链式反应(PCR)是分子生物学领域中检测极少量DNA和RNA样品的一种有用的工具。本研究的目的是用原位逆转录聚合酶链式反应(RT-PCR)方法对肾外组织中肾素表达细胞进行检测和鉴定，并用杂交组织化学、去势和免疫电子显微镜技术阐明肾素凝固腺的功能。在前者中，改变了探针、消化、RT和PCR反应的条件，包括溶液浓度和几次反应时间。结果发现，对大鼠的淋巴结有微弱的反应。它们仅限于毛细血管后小静脉。在后者中，凝固腺肾素以外分泌方式分泌到精腔，其表达受睾酮的调节。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，尽管该方法至今仍在进步，而且凝固性腺体肾素可能通过血管紧张素原的合成对女性生殖器官产生影响。

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.