Immunocytochemical localization of angiotensin II receptor subtypes and angiotensin II with monoclonal antibodies in the rat adrenal gland
Immunocytochemical localization of angiotensin II receptor subtypes and angiotensin II with monoclonal antibodies in the rat adrenal gland
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DOI：
10.1016/s0167-0115(01)00278-6
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发表时间：
2001-09-15
影响因子：
--
通讯作者：
Imboden, H
中科院分区：
文献类型：
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作者：
Frei, N;Weissenberger, J;Imboden, H
血管紧张素II（Angiotensin II，Ang II）是心血管功能和体液平衡的主要调节剂，通过两种亚型的G蛋白偶联受体（AT（1）和AT（2））介导其生物学作用。本研究的主要目的是制备抗血管紧张素AT（1）和AT（2）受体亚型及Ang II自身的单克隆抗肽抗体，并利用这些单克隆抗体确定AT（1）和AT（2）受体及Ang II在雄性成年大鼠肾上腺内的定位。免疫细胞化学明确显示了肾上腺中血管紧张素II和血管紧张素II受体的区域共定位。新抗体将血管紧张素II和AT（1）受体定位于皮质和髓质的肾小球，而AT（1）受体仅限于髓质。通过用免疫原性肽预吸附抗体来记录免疫染色的特异性。我们的数据强调AT（1）似乎介导了肾上腺中Ang II的大部分生理作用。使用AT（1）抗体对大鼠肾上腺蛋白提取物进行Western印迹分析，显示主要的73-kDa条带和较弱的97-kDa免疫反应条带，对应于AT（1）受体的糖基化形式。抗AT（2）免疫染色产生一条73 kDa的主要免疫反应带和一条120 kDa的较弱带。这些抗体可能在阐明AT（1）和AT（2）的亚细胞定位和细胞内效应途径方面具有价值。(C)2001 Elsevier Science B. V.保留所有权利。
Angiotensin II (Ang II), a major regulator of cardiovascular function and body fluid homeostasis, mediates its biological actions via two subtypes of G protein-coupled receptors, termed AT(1) and AT(2). The primary goal of this study was to raise monoclonal anti-peptide antibodies specific to angiotensin AT(1)- and AT(2)-receptor subtypes and to Ang II itself and using these monoclonal antibodies to determine the intraadrenal localization of AT(1) and AT(2) receptors and Ang II in male adult rats. Immunocytochemistry unambiguously demonstrates a regional colocalization of Ang II and angiotensin II receptors in the adrenal gland. The novel antibodies localized Ang II and the AT(1) receptors to the zona glomerulosa of the cortex and to the medulla whereas AT(1) receptors were limited to the medulla. The specificity of immunostaining was documented by pre-adsorption of the antibody with the immunogenic peptide. Our data underscore that AT(1) appears to mediate most of the physiological actions of Ang II in adrenal. Western blot analysis of rat adrenal protein extracts using AT(1) antibody showed a predominant 73-kDa band and a weaker 97-kDa immunoreactive band corresponding to glycosylated forms of the AT(1) receptor. Immunostaining with anti-AT(2) yielded one major immunoreactive band of 73-kDa size and one additional fainter band of 120 kDa. These antibodies may prove of value in unraveling the subcellular localization and intracellular effector pathways of AT(1) and AT(2). (C) 2001 Elsevier Science B.V. All rights reserved.