Regulation of glomerulotubular balance. III. Implication of cytosolic calcium in flow-dependent proximal tubule transport

Abstract

In the proximal tubule, axial flow (drag on brush-border microvilli) stimulates Na+ and HCO3− reabsorption by modulating both Na/H exchanger 3 (NHE3) and H-ATPase activity, a process critical to glomerulotubular balance. We have also demonstrated that blocking the angiotensin II receptor decreases baseline transport, but preserves the flow effect; dopamine leaves baseline fluxes intact, but abrogates the flow effect. In the current work, we provide evidence implicating cytosolic calcium in flow-dependent transport. Mouse proximal tubules were microperfused in vitro at perfusion rates of 5 and 20 nl/min, and reabsorption of fluid ( Jv) and HCO3− ( JHCO3) were measured. We examined the effect of high luminal Ca2+ (5 mM), 0 mM Ca2+, the Ca2+ chelator BAPTA-AM, the inositol 1,4,5-trisphosphate (IP3) receptor antagonist 2-aminoethoxydiphenyl borate (2-APB), and the Ca-ATPase inhibitor thapsigargin. In control tubules, increasing perfusion rate from 5 to 20 nl/min increased Jv by 62% and JHCO3 by 104%. With respect to Na+ reabsorption, high luminal Ca2+ decreased transport at low flow, but preserved the flow-induced increase; low luminal Ca2+ had little impact; both BAPTA and 2-APB had no effect on baseline flux, but abrogated the flow effect; thapsigargin decreased baseline flow, leaving the flow effect intact. With respect to HCO3− reabsorption, high luminal Ca2+ decreased transport at low flow and mildly diminished the flow-induced increase; low luminal Ca2+ had little impact; both BAPTA and 2-APB had no effect on baseline flux, but abrogated the flow effect. These data implicate IP3 receptor-mediated intracellular Ca2+ signaling as a critical step in transduction of microvillous drag to modulate Na+ and HCO3− transport.