Andreasen D, Jensen B L, Hansen P B, Kwon T H, Nielsen S, Skøtt O
Department of Physiology and Pharmacology, University of Southern Denmark-Odense University, DK-5000 Odense, Denmark.
Am J Physiol Renal Physiol. 2000 Dec;279(6):F997-1005. doi: 10.1152/ajprenal.2000.279.6.F997.
The molecular type and localization of calcium channels along the nephron are not well understood. In the present study, we assessed the distribution of the recently identified alpha(1G)-subunit encoding a voltage-dependent calcium channel with T-type characteristics. Using a RNase protection assay, alpha(1G)-mRNA levels in kidney regions were determined as inner medulla >> outer medulla congruent with cortex. RT-PCR analysis of microdissected rat nephron segments revealed alpha(1G) expression in the distal convoluted tubule (DCT), in the connecting tubule and cortical collecting duct (CT+CCD), and inner medullary collecting duct (IMCD). alpha(1G) mRNA was expressed in the IMCD cell line mIMCD-3. Single- and double-labeling immunohistochemistry and confocal laser microscopy on semithin paraffin sections of rat kidneys by using an anti-alpha(1G) antibody demonstrated a distinct labeling at the apical plasma membrane domains of DCT cells, CT principal cells, and IMCD principal cells.
肾单位中钙通道的分子类型和定位尚未完全明确。在本研究中,我们评估了最近鉴定出的编码具有T型特征的电压依赖性钙通道的α(1G)-亚基的分布。使用核糖核酸酶保护分析,确定肾脏各区域的α(1G)-mRNA水平为内髓>>外髓与皮质相当。对显微切割的大鼠肾单位节段进行逆转录聚合酶链反应分析显示,α(1G)在远曲小管(DCT)、连接小管和皮质集合管(CT+CCD)以及内髓集合管(IMCD)中表达。α(1G) mRNA在IMCD细胞系mIMCD-3中表达。通过使用抗α(1G)抗体对大鼠肾脏半薄石蜡切片进行单标记和双标记免疫组织化学及共聚焦激光显微镜检查,发现在DCT细胞、CT主细胞和IMCD主细胞的顶端质膜区域有明显标记。