title: Adenosine induces vasoconstriction through Gi-dependent activation of phospholipase C in isolated perfused afferent arterioles of mice.
abstract: Adenosine induces vasoconstriction of renal afferent arterioles through activation of A1 adenosine receptors (A1AR). A1AR are directly coupled to Gi/Go, resulting in inhibition of adenylate cyclase, but the contribution of this signaling pathway to smooth muscle cell activation is unclear. In perfused afferent arterioles from mouse kidney, adenosine and the A1 agonist N(6)-cyclohexyladenosine, when added to the bath, caused constriction in the concentration range of 10(-9) to 10(-6) M (mean diameter: control, 8.8 +/- 0.3 micro m; adenosine at 10(-6) M, 2.8 +/- 0.5 micro m). Adenosine-induced vasoconstriction was stable for up to 30 min and was most pronounced in the most distal part of the afferent arterioles. Adenosine did not cause vasoconstriction in arterioles from A1AR-/- mice. Pretreatment with pertussis toxin (PTX) (400 ng/ml) for 2 h blocked the vasoconstricting action of adenosine or N(6)-cyclohexyladenosine. PTX pretreatment did not affect the constriction response to KCl, whereas the angiotensin II dose-response relationship was shifted rightward. Reverse transcription-PCR revealed expression of Gi but not Go in kidney cortex and preglomerular vessels. The phospholipase C inhibitor U73122 (4 micro M) blocked the constriction responses to both adenosine and angiotensin II. In contrast, the adenylate cyclase inhibitor SQ22536 (10 micro M) and the protein kinase A antagonist KT5720 (0.1 and 1 micro M) did not induce significant vasoconstriction of afferent arterioles. It is concluded that the constriction response to adenosine in afferent arterioles is mediated by A1AR coupled to a PTX-sensitive Gi protein and subsequent activation of phospholipase C, presumably through betagamma subunits released from Galphai.
mesh_terms: D000241:Adenosine / Q000031:analogs & derivatives / Q000494:pharmacology*; D000818:Animals; D001160:Arterioles / Q000187:drug effects / Q000502:physiology; D005260:Female; D019206:GTP-Binding Protein alpha Subunits, Gi-Go / Q000378:metabolism*; D066298:In Vitro Techniques; D007678:Kidney Glomerulus / Q000098:blood supply*; D008297:Male; D051379:Mice; D008817:Mice, Mutant Strains; D010477:Perfusion; D037342:Pertussis Toxin / Q000494:pharmacology; D010738:Type C Phospholipases / Q000378:metabolism*; D014661:Vasoconstriction / Q000187:drug effects*; D014665:Vasodilator Agents / Q000494:pharmacology*
chemical_list: D014665:Vasodilator Agents; C027513:N(6)-cyclohexyladenosine; D037342:Pertussis Toxin; D010738:Type C Phospholipases; D019206:GTP-Binding Protein alpha Subunits, Gi-Go; D000241:Adenosine