Cellular localization of the potassium channel Kir7.1 in guinea pig and human kidney.

BACKGROUND K(+) channels have important functions in the kidney, such as maintenance of the membrane potential, volume regulation, recirculation, and secretion of potassium ions. The aim of this study was to obtain more information on the localization and possible functional role of the inwardly rectifying K(+) channel, Kir7.1. METHODS Kir7.1 cDNA (1114 bp) was isolated from guinea pig kidney (gpKir7.1), and its tissue distribution was analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR). In addition, a genomic DNA fragment (6153 bp) was isolated from a genomic library. cRNA was expressed in Xenopus laevis oocytes for functional studies. Immunohistochemistry and RT-PCR were used to localize Kir7.1 in guinea pig and human kidney. RESULTS The expression of gpKir7.1 in Xenopus laevis oocytes revealed inwardly rectifying K(+) currents. The reversal potential was strongly dependent on the extracellular K(+) concentration, shifting from -14 mV at 96 mmol/L K(+) to -90 mV at 1 mmol/L K(+). gpKir7.1 showed a low affinity for Ba(2+). Significant expression of gpKir7.1 was found in brain, kidney, and lung, but not in heart, skeletal muscle, liver, or spleen. Immunocytochemical detection in guinea pig identified the gpKir7.1 protein in the basolateral membrane of epithelial cells of the proximal tubule. RT-PCR analysis identified strong gpKir7.1 expression in the proximal tubule and weak expression in glomeruli and thick ascending limb. In isolated human tubule fragments, RT-PCR showed expression in proximal tubule and thick ascending limb. CONCLUSION Our results suggest that Kir7.1 may contribute to basolateral K(+) recycling in the proximal tubule and in the thick ascending limb.

[1]  A. Karschin,et al.  Genomic structure and promoter analysis of the rat Kir7.1 potassium channel gene (Kcnj13) , 2000, FEBS letters.

[2]  S. Tucker,et al.  pH Dependence of the Inwardly Rectifying Potassium Channel, Kir5.1, and Localization in Renal Tubular Epithelia* , 2000, The Journal of Biological Chemistry.

[3]  E. Schlatter,et al.  A novel cGMP‐regulated K+ channel in immortalized human kidney epithelial cells (IHKE‐1) , 1999, The Journal of physiology.

[4]  Y. Suzuki,et al.  Inwardly rectifying K+ channel Kir7.1 is highly expressed in thyroid follicular cells, intestinal epithelial cells and choroid plexus epithelial cells: implication for a functional coupling with Na+,K+-ATPase. , 1999, The Biochemical journal.

[5]  J. Daut,et al.  Separation of cardiomyocytes and coronary endothelial cells for cell-specific RT-PCR. , 1999, American journal of physiology. Heart and circulatory physiology.

[6]  M. Lazdunski,et al.  Expression of TWIK-1, a novel weakly inward rectifying potassium channel in rat kidney. , 1998, American journal of physiology. Cell physiology.

[7]  R. Schneggenburger,et al.  The Epithelial Inward Rectifier Channel Kir7.1 Displays Unusual K+ Permeation Properties , 1998, The Journal of Neuroscience.

[8]  I. Housini,et al.  Localization of the ROMK potassium channel to the apical membrane of distal nephron in rat kidney. , 1998, Kidney international.

[9]  Jürgen Lindemeier,et al.  Haptenylation of antibodies during affinity purification: a novel and convenient procedure to obtain labeled antibodies for quantification and double labeling , 1998, Histochemistry and Cell Biology.

[10]  D. Clapham,et al.  A Novel Inward Rectifier K+ Channel with Unique Pore Properties , 1998, Neuron.

[11]  A. S. Segal,et al.  Regulation of an Inwardly Rectifying ATP-sensitive K+ Channel in the Basolateral Membrane of Renal Proximal Tubule , 1998, The Journal of general physiology.

[12]  J. Wade,et al.  Localization of ROMK channels in the rat kidney. , 1997, Journal of the American Society of Nephrology : JASN.

[13]  P. Welling,et al.  Basolateral membrane targeting of a renal-epithelial inwardly rectifying potassium channel from the cortical collecting duct, CCD-IRK3, in MDCK cells. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[14]  S. Hebert,et al.  Localization of the ROMK protein on apical membranes of rat kidney nephron segments. , 1997, American journal of physiology. Renal physiology.

[15]  M. L. Watkins,et al.  A simplified method for isolation of large numbers of defined nephron segments. , 1997, American journal of physiology. Renal physiology.

[16]  H. Choe,et al.  Is the secretory K channel in the rat CCT ROMK? , 1997, The American journal of physiology.

[17]  J. Slightom,et al.  Cloning and Characterization of Two K+ Inward Rectifier (Kir) 1.1 Potassium Channel Homologs from Human Kidney (Kir1.2 and Kir1.3)* , 1997, The Journal of Biological Chemistry.

[18]  F. Ortis,et al.  Immunopurification of polyclonal antibodies to recombinant proteins of the same gene family. , 1996, BioTechniques.

[19]  R. Lifton,et al.  Genetic heterogeneity of Barter's syndrome revealed by mutations in the K+ channel, ROMK , 1996, Nature Genetics.

[20]  Y. Horio,et al.  Immunolocalization of an inwardly rectifying K+ channel, KAB‐2 (Kir4.1), in the basolateral membrane of renal distal tubular epithelia , 1996, FEBS letters.

[21]  S. Hebert An ATP-regulated, inwardly rectifying potassium channel from rat kidney (ROMK). , 1995, Kidney international.

[22]  W. Wang,et al.  Two types of K+ channel in thick ascending limb of rat kidney. , 1994, The American journal of physiology.

[23]  G. Giebisch,et al.  ATP is a coupling modulator of parallel Na,K-ATPase-K-channel activity in the renal proximal tubule. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[24]  R. Sauvé,et al.  Hypotonic shock activates a maxi K+ channel in primary cultured proximal tubule cells. , 1990, American Journal of Physiology.

[25]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[26]  W. Guggino,et al.  Ca2+-activated K+ channels in cultured medullary thick ascending limb cells. , 1987, The American journal of physiology.

[27]  M. Burg,et al.  Thick ascending limb of Henle's loop. , 1982, Kidney international.

[28]  A. Ziegler,et al.  Mutations in the gene encoding the inwardly-rectifying renal potassium channel, ROMK, cause the antenatal variant of Bartter syndrome: evidence for genetic heterogeneity. International Collaborative Study Group for Bartter-like Syndromes. , 1997, Human molecular genetics.

[29]  F. Lang,et al.  Potassium channels in renal epithelial transport regulation. , 1992, Physiological reviews.

[30]  E. Harlow,et al.  Antibodies: A Laboratory Manual , 1988 .