title: Bimodal acute effects of A1 adenosine receptor activation on Na+/H+ exchanger 3 in opossum kidney cells.
abstract: Regulation of renal apical Na+/H+ exchanger 3 (NHE3) activity by adenosine has been suggested to contribute to acute control of mammalian Na(+) homeostasis. The mechanism by which adenosine controls NHE3 activity in a renal cell line was examined. The adenosine analog, N(6)-cyclopentyladenosine (CPA) exerts a bimodal effect on NHE3: CPA concentrations >10(-8) M inactivate NHE3, whereas concentrations <10(-8) M stimulate NHE3 activity. Acute CPA-induced control of NHE3 was blocked by antagonists of A1 adenosine receptors and inhibition of phospholipase C, pretreatment with BAPTA-AM (chelator of cellular calcium), and exposure to pertussis toxin. Stimulatory and to some extent also inhibitory CPA concentrations attenuated 8-bromo-cAMP and dopamine-mediated inhibition of NHE3. BAPTA eliminated the ability of a stimulatory dose of CPA to attenuate 8-bromo-cAMP-induced suppression of NHE3 activity. Upon inhibition of protein kinase C, CPA at an inhibitory dose provoked activation of NHE3, which is partially reverted by 8-bromo-cAMP and suppressed by pre-incubation with BAPTA-AM. Cytochalasin B, an actin-modifying agent, selectively prevented downregulation but did not affect upregulation of NHE3 activity by CPA. In conclusion, these observations demonstrate that (1) CPA modulates NHE3 activity by elevation of cellular Ca(2+) exerting a negative control on adenylate cyclase activity, (2) protein kinase C is the determining factor leading to CPA-induced downregulation of NHE3 activity, and (3) alterations of surface NHE3 abundance may contribute to A1 adenosine receptor-dependent inhibition of NHE3 activity.
authors.affiliation: Department of Physiology and Pathophysiology, Division of Vegetative Physiology and Pathophysiology, Georg-August University of Goettingen, Humboldtallee 23, D-37073 Goettingen, Germany. sole@veg-physiol.med.uni-goettingen.de
authors.forename: Francesca
authors.initials: F
authors.lastname: Di Sole
authors.forename: Robert
authors.initials: R
authors.lastname: Cerull
authors.forename: Soeren
authors.initials: S
authors.lastname: Petzke
authors.forename: Valeria
authors.initials: V
authors.lastname: Casavola
authors.forename: Gerhard
authors.initials: G
authors.lastname: Burckhardt
authors.forename: Corinna
authors.initials: C
authors.lastname: Helmle-Kolb
mesh_terms: D000241:Adenosine / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002614:Chelating Agents / Q000494:pharmacology; D003571:Cytochalasin B / Q000494:pharmacology; D018076:DNA, Complementary / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D004533:Egtazic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology; D007668:Kidney / Q000166:cytology* / Q000378:metabolism; D009893:Opossums; D037342:Pertussis Toxin / Q000494:pharmacology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D017434:Protein Structure, Tertiary; D018047:Receptors, Purinergic P1 / Q000378:metabolism*; D015398:Signal Transduction; D012964:Sodium / Q000378:metabolism; D000074083:Sodium-Hydrogen Exchanger 3; D017923:Sodium-Hydrogen Exchangers / Q000378:metabolism*; D013997:Time Factors; D015854:Up-Regulation
chemical_list: D002614:Chelating Agents; D018076:DNA, Complementary; D018047:Receptors, Purinergic P1; D000074083:Sodium-Hydrogen Exchanger 3; D017923:Sodium-Hydrogen Exchangers; C070379:1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester; D003571:Cytochalasin B; C048599:N(6)-cyclopentyladenosine; D004533:Egtazic Acid; D012964:Sodium; D037342:Pertussis Toxin; D011493:Protein Kinase C; D000241:Adenosine; D002118:Calcium