Vandewalle A, Cluzeaud F, Bens M, Kieferle S, Steinmeyer K, Jentsch T J
Institut National de la Santé et de la Recherche Médicale Unité, Paris, France.
Am J Physiol. 1997 May;272(5 Pt 2):F678-88. doi: 10.1152/ajprenal.1997.272.5.F678.
We investigate the intrarenal expression of two recently cloned chloride channels, rClC-K1 and rClC-K2, by reverse transcriptase-polymerase chain reaction on single microdissected tubules from the rat kidney and by immunohistochemistry using a polyclonal antibody that recognizes both highly homologous channels. Both rClC-K1 and rClC-K2 mRNAs were detected in outer medullary late proximal tubules (S3), papillary ascending thin limbs (ATL), and outer medullary (MTAL) and cortical (CTAL) thick ascending limbs, distal tubules (DCT), and cortical, outer medullary, and inner medullary collecting ducts. Indirect immunofluorescence studies demonstrated that the rClC-K proteins were restricted to the basolateral membranes from ATL, DCT, and collecting ducts cells, whereas CTAL and MTAL exhibited a more diffuse basal staining. When rats were dehydrated, a condition which increased the expression of rClC-K1 in cortex and medulla, a weak cytoplasmic staining was found in late proximal tubule cells. Thus these results demonstrate that rat kidney ClC-K channels are predominantly located in the basolateral membranes from cells of the late segments of the renal tubule where most of chloride reabsorption takes place.
我们通过逆转录聚合酶链反应,对大鼠肾脏单个显微切割小管进行研究,以检测两种最近克隆的氯通道rClC-K1和rClC-K2的肾内表达,并使用一种能识别这两种高度同源通道的多克隆抗体进行免疫组织化学检测。在髓质外层近端小管晚期(S3)、乳头上升细段(ATL)、髓质外层(MTAL)和皮质(CTAL)厚上升段、远端小管(DCT)以及皮质、髓质外层和髓质内层集合管中均检测到rClC-K1和rClC-K2 mRNA。间接免疫荧光研究表明,rClC-K蛋白局限于ATL、DCT和集合管细胞的基底外侧膜,而CTAL和MTAL则呈现更弥散的基底染色。当大鼠脱水时,这种情况会增加皮质和髓质中rClC-K1的表达,此时在近端小管晚期细胞中发现了微弱的细胞质染色。因此,这些结果表明,大鼠肾脏ClC-K通道主要位于肾小管晚期段细胞的基底外侧膜,而大部分氯离子重吸收发生在此处。