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Immunocytochemistry and in situ hybridization of some vasoactive peptides and their receptors

一些血管活性肽及其受体的免疫细胞化学和原位杂交

基本信息

批准号：
09670035
负责人：
FUJIMOTO Sunao
金额：
$ 1.41万
依托单位：
University of Occupational and Environmental Health
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

项目摘要

1)The rat carotid body artery (CBA) and thoracic aorta under the hypoxic condition were investigated by electron microscopy, immunocytochernistry and in-situ hybridization. A remarkable increase in the number of Weibel-Palade (WP) bodies occurred in endothelial cells of the hypoxic CBA and thoracic aorta. The degree of immunoreactivities and gene expressions of both endothelin (ET)-1 and calcitonin gene-related peptide (CGRP) was significantly enhanced in the hypoxic samples when compared to the normoxic ones. Since WP bodies are involved in the storage and release of these peptides, the present findings indicate that ET-1 and CGRP, which are actively synthesized and released from endothelial cells, play a crucial role in the regulation of the blood flow through these vessels in the hypoxic condition.2)Vasoactivities after treatment with ET-1, and immunoreactivities for ET receptors were investigated on the rat superior mesenteric vasculature (SMV). By the me asurement of corrosion cast images of the SMVs, it was seen that ET-1. induces the remarkable vasocontraction of the distal arterial branches. The present immunocytochemistry showed that immunoreactivities for ETA receptors are localized to the sarcolemma of medial muscle, cells of the distal arterial branches. These findings suggest that ET-1-induced vasocontraction on the distal arterial branches is mediated from the ETA receptors. Immunoreactivities for ETB receptors were seen on the plasmalemma of endothelial cells in the SMVs. Since the WP bodies of these endothelial cells showed ultrastructural changes such as degranulation and exocytosis after treatment with sarafotoxin S6c, ETB receptor selective agonist, it seems likely that ETB receptors of the SMVs are involved in the release of the WP bodies.

1)采用电镜、免疫细胞化学和原位杂交技术对缺氧状态下的大鼠颈动脉体(CBA)和胸主动脉进行观察。缺氧的 CBA 和胸主动脉的内皮细胞中 Weibel-Palade (WP) 小体的数量显着增加。与常氧样品相比，低氧样品中内皮素 (ET)-1 和降钙素基因相关肽 (CGRP) 的免疫反应性程度和基因表达均显着增强。由于WP体参与这些肽的储存和释放，目前的研究结果表明，内皮细胞主动合成和释放的ET-1和CGRP在缺氧条件下调节通过这些血管的血流中发挥着至关重要的作用。2)在大鼠肠系膜上血管系统上研究了ET-1治疗后的血管活性和ET受体的免疫反应性（SMV）。通过对SMV的腐蚀铸件图像的测量，可以看出ET-1。引起远端动脉分支显着的血管收缩。目前的免疫细胞化学显示，ETA 受体的免疫反应性位于内侧肌肉的肌膜、远端动脉分支的细胞。这些发现表明，ET-1 诱导的远端动脉分支血管收缩是由 ETA 受体介导的。在 SMV 内皮细胞的质膜上观察到 ETB 受体的免疫反应性。由于这些内皮细胞的 WP 小体在用 ETB 受体选择性激动剂 sarafotoxin S6c 处理后显示出超微结构变化，例如脱颗粒和胞吐作用，因此 SMV 的 ETB 受体似乎参与了 WP 小体的释放。