Immunolocalization of V-1 vasopressin receptors in the rat kidney using anti-receptor antibodies
Immunolocalization of V-1 vasopressin receptors in the rat kidney using anti-receptor antibodies
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DOI：
10.1038/ki.1997.445
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发表时间：
1997-11-01
影响因子：
19.6
通讯作者：
Menzel, D
中科院分区：
文献类型：
--
作者：
Gonzalez, CB;Figueroa, CD;Menzel, D
利用免疫细胞化学技术，我们已经能够定位的V-1加压素受体在大鼠肾脏。免疫印迹使用抗血清提出的亲和纯化的加压素受体显示了55，000道尔顿的蛋白质带，具有类似的分子量的肝V-1加压素受体，如交联研究所证明的。抗体的免疫印迹显示在A-10细胞中的55，000道尔顿的条带，其含有V-1亚型，而其不染色具有V-2亚型的LLC-PK 1细胞，表明抗体识别V-1加压素受体。用该抗血清对肾切片进行免疫染色，显示连接小管和皮质及髓质集合管的强烈反应。连接小管和集合管细胞的免疫染色模式是不同的，即，前者表现出染色的顶端和基底质膜，但也在细胞质中，而后者表现出强烈的反应，主要是在基底侧膜。用抗组织激肽释放酶（一种仅存在于连接小管中的酶）的抗血清和抗受体血清对连续连续切片进行免疫染色，使我们能够首次显示连接小管细胞中存在加压素受体，而在闰细胞中不存在加压素受体，闰细胞是连接小管中存在的另一种细胞类型。这些发现支持了80年代进行的关于AVP释放肾组织激肽释放酶的实验。
By using immunocytochemical techniques we have been able to localize the V-1 vasopressin receptor in the rat kidney. Immunoblotting using an antiserum raised against an affinity-purified vasopressin receptor showed a 55,000 daltons protein band that has a molecular mass similar to that of the liver V-1 vasopressin receptor, as demonstrated by cross-linking studies. Immunoblotting of the antibody showed a band of 55,000 daltons in A-10 cells, which contains the V-1 subtype, whereas it did not stain LLC-PK1 cells, which possess the V-2 subtype, showing that the antibody recognizes the V-1 vasopressin receptor. The immunostaining of kidney sections with this antiserum showed a strong reaction of the connecting tubules and cortical and medullary collecting ducts. The immunostaining pattern of connecting tubule and collecting duct cells was different, that is, the former showed a staining of both the apical and basal plasma membrane but also in the cytoplasm, whereas the latter showed a strong reaction mainly in the basolateral membrane. Immunostaining of consecutive serial sections with an antiserum raised against tissue kallikrein, an enzyme present exclusively in connecting tubules, and with the anti-receptor serum allowed us to show, for the first time, the presence of the vasopressin receptor in the connecting tubule cells and their absence in intercalated cells, the other cell type present in connecting tubules. These findings support experiments carried in the eighties on the release of renal tissue kallikrein by AVP.