title: Protein kinase G inhibits flow-induced Ca2+ entry into collecting duct cells.
abstract: The renal cortical collecting duct (CCD) contributes to the maintenance of K(+) homeostasis by modulating renal K(+) secretion. Cytosolic Ca(2+) ([Ca(2+)](i)) mediates flow-induced K(+) secretion in the CCD, but the mechanisms regulating flow-induced Ca(2+) entry into renal epithelial cells are not well understood. Here, we found that atrial natriuretic peptide, nitric oxide, and cyclic guanosine monophosphate (cGMP) act through protein kinase G (PKG) to inhibit flow-induced increases in [Ca(2+)](i) in M1-CCD cells. Coimmunoprecipitation, double immunostaining, and functional studies identified heteromeric TRPV4-P2 channels as the mediators of flow-induced Ca(2+) entry into M1-CCD cells and HEK293 cells that were coexpressed with both TRPV4 and TRPP2. In these HEK293 cells, introducing point mutations at two putative PKG phosphorylation sites on TRPP2 abolished the ability of cGMP to inhibit flow-induced Ca(2+) entry. In addition, treating M1-CCD cells with fusion peptides that compete with the endogenous PKG phosphorylation sites on TRPP2 also abolished the cGMP-mediated inhibition of the flow-induced Ca(2+) entry. Taken together, these data suggest that heteromeric TRPV4-P2 channels mediate the flow-induced entry of Ca(2+) into collecting duct cells. Furthermore, substances such as atrial natriuretic peptide and nitric oxide, which increase cGMP, abrogate flow-induced Ca(2+) entry through PKG-mediated inhibition of these channels.
authors.affiliation: School of Biomedical Sciences, Chinese University of Hong Kong, China.
authors.forename: Juan
authors.initials: J
authors.lastname: Du
authors.forename: Wei-Yan
authors.initials: WY
authors.lastname: Wong
authors.forename: Lei
authors.initials: L
authors.lastname: Sun
authors.forename: Yu
authors.initials: Y
authors.lastname: Huang
authors.forename: Xiaoqiang
authors.initials: X
authors.lastname: Yao
mesh_terms: D000818:Animals; D009320:Atrial Natriuretic Factor / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002460:Cell Line; D006152:Cyclic GMP / Q000494:pharmacology; D017869:Cyclic GMP-Dependent Protein Kinases / Q000187:drug effects / Q000494:pharmacology*; D057809:HEK293 Cells; D006801:Humans; D066298:In Vitro Techniques; D007685:Kidney Tubules, Collecting / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D051379:Mice; D023421:Models, Animal; D009569:Nitric Oxide / Q000494:pharmacology; D010766:Phosphorylation / Q000187:drug effects; D050052:TRPC Cation Channels / Q000187:drug effects / Q000502:physiology; D050396:TRPP Cation Channels / Q000187:drug effects / Q000502:physiology; D050916:TRPV Cation Channels / Q000187:drug effects / Q000502:physiology
chemical_list: D009320:Atrial Natriuretic Factor; D002118:Calcium; D006152:Cyclic GMP; D017869:Cyclic GMP-Dependent Protein Kinases; D009569:Nitric Oxide; D050052:TRPC Cation Channels; D050396:TRPP Cation Channels; D050916:TRPV Cation Channels; C416986:TRPV4 protein, human; C486576:Trpv4 protein, mouse; C096351:transient receptor potential cation channel, subfamily C, member 1