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J Biol Chem, Vol. 273, Issue 24, 14906-14911, June 12, 1998

Phosphatidylinositol 3-Kinase Contributes to Cell Volume Regulation through Effects on ATP Release

From the Departments of Pediatrics and Medicine, Children's Hospital and the University of Colorado Health Sciences Center, Denver, Colorado 80262 and the ^§ Department of Physiology and Biophysics, University of Alabama, Birmingham, Alabama 35294

Regulated changes in cell volume represent a signal that modulates a broad range of cell and organ functions. In HTC hepatoma cells, increases in volume are coupled to membrane ion permeability through a pathway involving (i) ATP efflux, (ii) autocrine stimulation of P₂ receptors, and (iii) increases in anion permeability and Cl efflux, contributing to recovery of volume toward basal values. Based on recent evidence that cell volume increases also stimulate phosphoinositide kinases, the purpose of these studies was to determine if phosphatidylinositol 3-kinase (PI 3-kinase) modulates these pathways. Exposure of cells to hypotonic buffer (20 or 40% less NaCl) caused an initial increase in cell volume and stimulated a rapid increase in ATP release. Subsequent opening of Cl channels was followed by recovery of cell volume toward basal values, despite the continuous presence of hypotonic buffer. Inhibition of PI 3-kinase with wortmannin (K_i = 3 nM) significantly inhibited both the rate of volume recovery and activation of Cl currents; similar results were obtained with LY294002 (10 µM). Additionally, current activation was inhibited by intracellular dialysis with antibodies specific for the 110-kDa catalytic subunit of PI 3-kinase. Since release of ATP is a critical element in the volume-regulatory pathway, the role of PI 3-kinase on volume-stimulated ATP release was assessed. Both wortmannin and LY294002 decreased basal and volume-stimulated ATP permeability but had no effect on the current response to exogenous ATP (10 µM). These findings indicate that PI 3-kinase plays a significant role in regulation of cell volume and suggest that the effects are mediated in part through modulation of cellular ATP release.

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