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Inhibition of bicarbonate reabsorption in the rat proximal tubule by activation of luminal P2Y₁ receptors

Department of Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, Connecticut 06520

Submitted 2 February 2004 ; accepted in final form 27 May 2004

The present study used a stationary microperfusion technique to investigate in vivo the effect of P2Y₁ receptor activation on bicarbonate reabsorption in the rat proximal tubule. Proximal tubules were perfused with a bicarbonate Ringer solution before flow was stopped by means of an oil block. The recovery of lumen pH from the initial value (pH 8.0) to stationary values (pH 6.7) was recorded by a H⁺-sensitive microelectrode inserted downstream of the perfusion pipette and oil block. The stationary pH value and the t of pH recovery were used to calculate bicarbonate reabsorption (J_HCO3). Both EIPA and bafilomycin A1 caused significant reductions in proximal tubule J_HCO3, consistent with the established contributions of Na/H exchange and H⁺-ATPase to proximal tubule HCO₃ reabsorption. The nucleotides ADP and, to a lesser extent, ATP reduced J_HCO3 but AMP and UTP were without effect. 2MeSADP, a highly selective agonist of the P2Y₁ receptor, reduced J_HCO3 in a dose-dependent manner. MRS-2179, a P2Y₁ receptor-specific antagonist, abolished the effect of 2MeSADP, whereas theophylline, an antagonist of adenosine (P1) receptors, did not. The inhibitory action of 2MeSADP was blocked by inhibition of protein kinase C and reduced by inhibition of protein kinase A. The effects of EIPA and 2MeSADP were not additive. The data provide functional evidence for P2Y₁ receptors in the apical membrane of the rat proximal tubule: receptor activation impairs acidification in this nephron segment.

P2 receptor; purinergic; renal; extracellular nucleotide

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