Localization of the thiazide sensitive Na-Cl cotransporter, rTSC1 in the rat kidney.
Localization of the thiazide sensitive Na-Cl cotransporter, rTSC1 in the rat kidney.
复制标题
噻嗪类敏感的 Na-Cl 协同转运蛋白 rTSC1 在大鼠肾脏中的定位。
DOI：
10.1038/ki.1996.300
复制
发表时间：
1996
影响因子：
19.6
通讯作者：
Hebert,SC
中科院分区：
文献类型：
--
作者：
Plotkin,MD;Kaplan,MR;Verlander,JW;Lee,WS;Brown,D;Poch,E;Gullans,SR;Hebert,SC
噻嗪敏感Na-Cl共转运体rTSC1在大鼠肾脏中的定位。一个噻嗪敏感的Na-C1共转运蛋白rTSC1最近从大鼠肾皮质cDNA文库中克隆出来。该蛋白的分子调控和肾细胞定位尚不清楚。本研究的目的是研究rTSC1蛋白在大鼠肾脏中的分布和亚细胞定位。原位杂交显示rTSC1转录本定位于肾皮质的短而卷曲的小管片段。针对rTSC1氨基酸端110个氨基酸片段的多克隆抗体在大鼠肾皮质而非外髓质膜蛋白的Western blotting上识别出135、140和155 kDa的三个主要条带。免疫荧光研究单独使用抗体和与H+ATPase和calbindin D28抗体的双标记实验显示，远端肾元顶端膜的强烈标记始于初始远端曲小管，终止于连接小管。标记强度沿远端小管从近端到远端部位减弱。免疫电镜超微结构研究显示，该共转运蛋白主要定位于远曲小管细胞的顶端微绒毛上。这些结果与rTSC1编码远端小管顶端噻嗪类敏感Na-Cl共转运蛋白的观点一致。
Localization of the thiazide sensitive Na-Cl cotransporter, rTSC1, in the rat kidney.A thiazide sensitive Na-C1 cotransporter, rTSC1, has recently been cloned from a rat kidney cortex cDNA library. The molecular regulation and nephron localization of this protein is unknown. The purpose of this study was to examine the nephron distribution and subcellular localization of the rTSC1 protein in the rat kidney.In situhybridization showed rTSC1 transcripts were localized to short, convoluted tubule segments in the kidney cortex. Polyclonal antibodies raised against a 110 amino acid segment from the amino terminus of rTSC1 recognized three major bands of 135, 140 and 155 kDa on Western blotting of membrane protein from cortex but not outer medulla of the rat kidney. Immunofluorescence studies using the antibody alone and in double labeling experiments with antibodies against the H+ATPase and calbindin D28, showed intense labeling of apical membranes in the distal nephron beginning in the initial distal convoluted tubule and terminating within the connecting tubule. The intensity of labeling diminished from proximal to distal sites along the distal tubule. Ultrastructural studies by immunoelectron microscopy showed the cotransporter protein to be localized predominately on apical microvilli of the distal convoluted tubule cells. These results are consistent with rTSC1 encoding the apical thiazide sensitive Na-Cl cotransporter in the distal tubule.