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Arginine vasopressin stimulates H⁺-ATPase in MDCK cells via V₁ (cell Ca²⁺) and V₂ (cAMP) receptors

Department of Physiology and Biophysics, Instituto de Ciências Biomédicas, University of São Paulo, São Paulo 05508-900, Brazil

Submitted 25 March 2003 ; accepted in final form 26 August 2003

The effect of arginine vasopressin (AVP) and/or atrial natriuretic peptide (ANP) on the regulation of intracellular pH (pH_i) via H⁺-ATPase and of cytosolic calcium ([Ca²⁺]_i) was investigated in Madin-Darby canine kidney (MDCK) cells by the fluorescent probes BCECF-AM and fluo-4-AM, respectively. The pH_i recovery rate was examined after intracellular acidification following an NH₄Cl pulse, in the presence of zero Na⁺ plus Schering 28080 (a specific inhibitor of H⁺-K⁺-ATPase). AVP (10^-12-10^-6 M) increased the rate of pH_i recovery and [Ca²⁺]_i in a dose-dependent manner. V₁- or V₂-receptor antagonists impaired the effect of AVP on both processes, and DDAVP (10^-12-10^-6 M; a V₂-selective agonist) caused a dose-dependent stimulation of them. [Ca²⁺]_i or cAMP (as increased by 10^-5 M thapsigargin or 8-BrcAMP, respectively) alone had no effect on H⁺-ATPase, but their synergic action was necessary to stimulate H⁺-ATPase. In agreement with these findings, ANP (10^-6 M) or dimethyl-BAPTA-AM (5 x 10^-5 M), impairing the increase of [Ca²⁺]_i in response to AVP, blocks the stimulatory effect of AVP on H⁺-ATPase.

atrial natriuretic peptide; Madin-Darby canine kidney cells

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