Identification and localization of TRPC channels in the rat kidney

被引:108
作者
Goel, M
Sinkins, WG
Zuo, CD
Estacion, M
Schilling, WP
机构
[1] Metrohlth Med Ctr, Rammelkamp Ctr Educ & Res, Cleveland, OH 44109 USA
[2] Case Western Reserve Univ, Sch Med, Dept Physiol & Biophys, Cleveland, OH 44106 USA
关键词
ion channels; renal nephron; immunoprecipitation; immunofluorescence; subcellular localization; polyclonal antibodies;
D O I
10.1152/ajprenal.00376.2005
中图分类号
Q4 [生理学];
学科分类号
071003 ;
摘要
It is well established that transient receptor potential (TRP) channels are activated following stimulation of G protein-coupled membrane receptors linked to PLC, but their differential expression in various cells of the renal nephron has not been described. In the present study, immunoprecipitations from rat kidney lysates followed by Western blot analysis using TRPC-specific, affinity-purified antibodies revealed the presence of TRPC1, -C3, and -C6. TRPC4, -C5, and -C7 were nondetectable. TRPC1 immunofluorescence was detected in glomeruli and specific tubular cells of the cortex and outer medulla. TRPC1 colocalized with aquaporin-1, a marker for proximal tubule and thin descending limb, but not with aquaporin-2, a marker for connecting tubule and collecting duct cells. TRPC3 and -C6 immunolabeling was predominantly confined to glomeruli and specific tubular cells of the cortex and both the outer and inner medulla. TRPC3 and -C6 colocalized with aquaporin-2, but not with the Na(+)/Ca(2+) exchanger or peanut lectin. Thus TRPC3 and -C6 proteins are expressed in principle cells of the collecting duct. In polarized cultures of M1 and IMCD-3 collecting duct cells, TRPC3 was localized exclusively to the apical domain, whereas TRPC6 was found in both the basolateral and apical membranes. TRPC3 and TRPC6 were also detected in primary podocyte cultures, whereas TRPC1 was exclusively expressed in mesangial cell cultures. Specific immunopositive labeling for TRPC4, -C5, or -C7 was not observed in kidney sections or cell lines. These results suggest that TRPC1, -C3, and -C6 may play a functional role in PLC-dependent signaling in specific regions of the nephron.
引用
收藏
页码:F1241 / F1252
页数:12
相关论文
共 41 条
[1]   Apical localization of a functional TRPC3/TRPC6-Ca2+-signaling complex in polarized epithelial cells -: Role in apical Ca2+ influx [J].
Bandyopadhyay, BC ;
Swaim, WD ;
Liu, XB ;
Redman, RS ;
Patterson, RL ;
Ambudkar, IS .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2005, 280 (13) :12908-12916
[2]   Distribution of transient receptor potential channels in the rat carotid chemosensory pathway [J].
Buniel, MCF ;
Schilling, WP ;
Kunze, DL .
JOURNAL OF COMPARATIVE NEUROLOGY, 2003, 464 (03) :404-413
[3]   Requirement for the PDZ domain protein, INAD, for localization of the TRP store-operated channel to a signaling complex [J].
Chevesich, J ;
Kreuz, AJ ;
Montell, C .
NEURON, 1997, 18 (01) :95-105
[4]   Signaling microdomains define the specificity of receptor-mediated InsP3 pathways in neurons [J].
Delmas, P ;
Wanaverbecq, N ;
Abogadie, FC ;
Mistry, M ;
Brown, DA .
NEURON, 2002, 34 (02) :209-220
[5]   Expression and relative abundance of short transient receptor potential channels in the rat renal microcirculation [J].
Facemire, CS ;
Mohler, PJ ;
Arendshorst, WJ .
AMERICAN JOURNAL OF PHYSIOLOGY-RENAL PHYSIOLOGY, 2004, 286 (03) :F546-F551
[6]   Specific detection and semi-quantitative analysis of TRPC4 protein expression by antibodies [J].
Flockerzi, V ;
Jung, C ;
Aberle, T ;
Meissner, M ;
Freichel, M ;
Philipp, SE ;
Nastainczyk, W ;
Maurer, P ;
Zimmermann, R .
PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY, 2005, 451 (01) :81-86
[7]   CA-2+-DEPENDENT INHIBITION OF SODIUM-TRANSPORT IN RABBIT CORTICAL COLLECTING TUBULES [J].
FRINDT, G ;
WINDHAGER, EE .
AMERICAN JOURNAL OF PHYSIOLOGY, 1990, 258 (03) :F568-F582
[8]   Differential expression of mammalian TRP homologues across tissues and cell lines [J].
Garcia, RL ;
Schilling, WP .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1997, 239 (01) :279-283
[9]   Selective association of TRPC channel subunits in rat brain synaptosomes [J].
Goel, M ;
Sinkins, WG ;
Schilling, WP .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2002, 277 (50) :48303-48310
[10]   INTERLEUKIN-1-ALPHA CAUSES RAPID ACTIVATION OF CYTOSOLIC PHOSPHOLIPASE A(2) BY PHOSPHORYLATION IN RAT MESANGIAL CELLS [J].
GRONICH, J ;
KONIECZKOWSKI, M ;
GELB, MH ;
NEMENOFF, RA ;
SEDOR, JR .
JOURNAL OF CLINICAL INVESTIGATION, 1994, 93 (03) :1224-1233