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Am J Physiol Renal Physiol (September 9, 2003). doi:10.1152/ajprenal.00121.2003
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Submitted on March 25, 2003
Accepted on August 26, 2003

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

Maria Oliveira-Souza1, Raif Musa-Aziz1, Gerhard Malnic1, and Margarida De Mello-Aires1*

1 Department of Physiology and Biophysics, Instituto de Ciencias Biomedicas, University of Sao Paulo, Sao Paulo, SP, Brazil

* To whom correspondence should be addressed. E-mail: mmaires{at}fisio.icb.usp.br.

The effect of Arginine Vasopressin (AVP) and/or Atrial Natriuretic Peptide (ANP) on the regulation of intracellular pH (pHi) 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. pHi recovery rate was examined following the intracellular acidification after an NH4Cl pulse, in presence of zero Na+ plus Schering 28080 (a specific inhibitor of H+/K+-ATPase). AVP (10-12 - 10-6 M) increased the rate of pHi 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-Br-cAMP, 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, block the stimulatory effect of AVP on H+-ATPase.




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