Ligands and signaling of the G-protein-coupled receptor GPR14, expressed in human kidney cells.
Activation of the urotensin II (U-II) receptor, GPR14, leads to an increase in Ca(2+), activation of phospholipase A(2) (PLA(2)) and an increase in arachidonic acid. The signaling pathway for guanylin peptides in the kidney involves an unknown G-protein coupled receptor which activates PLA(2) and increases arachidonic acid as well. To test if guanylin peptides could be, as U-II, agonists for the GPR14 receptor in the kidney, we used HEK293 and CHO cells transfected with hGPR14 (HEK293+hGPR14, CHO+hGPR14, respectively). Effects of guanylin peptides and U-II were studied by slow-whole-cell patch-clamp analysis and microfluorimetric measurements of intracellular Ca(2+). Guanylin peptides and U-II depolarized HEK293+hGPR14 significantly more than wild type cells. These effects were inhibited in the presence of Ba(2+) or PLA(2) inhibition (AACOCF(3)), suggesting that guanylin peptides and U-II increase arachidonic acid and inhibit ROMK channels in these cells. However, only U-II was capable to increase the cellular Ca(2+), suggesting different mechanism of GPR14 activation by guanylin peptides and U-II. This signaling pathway of U-II involves PKC, because U-II effects in HEK293+hGPR14 cells were inhibited by calphostin C. Guanylin peptides activate PLA(2) and inhibit ROMK channels in HEK293 cells transfected with the human GPR14 receptor. Since GPR14 is present in mouse and human CCD it is a candidate for the guanylate cyclase independent receptor for guanylin peptides.
Lehner
Ulrike
U
Universitätsklinikum Münster, Medizinische Klinik und Poliklinik D, Experimentelle Nephrologie, Münster, Germany.
Velić
Ana
A
Schroter
Rita
R
Schlatter
Eberhard
E
Sindić
Aleksandra
A
D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D020013:Calcium Signaling / Q000187:drug effects; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D017931:DNA Primers / Q000235:genetics; D005768:Gastrointestinal Hormones / Q000494:pharmacology; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000378:metabolism*; D008024:Ligands; D008564:Membrane Potentials / Q000187:drug effects; D045265:Natriuretic Peptides / Q000494:pharmacology; D010741:Phospholipases A / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D014162:Transfection; D014579:Urotensins / Q000494:pharmacology
D017931:DNA Primers; D005768:Gastrointestinal Hormones; D008024:Ligands; D045265:Natriuretic Peptides; D012333:RNA, Messenger; D043562:Receptors, G-Protein-Coupled; D011994:Recombinant Proteins; C121697:UTS2R protein, human; D014579:Urotensins; C072685:guanylin; C029677:urotensin II; D010741:Phospholipases A