Apical membrane sodium and chloride entry during osmotic swelling of renal (A6) epithelial cells.
To assess the role of chloride in cell volume and sodium transport regulation, we measured cell height changes (CH), transepithelial chloride and sodium fluxes, and intracellular chloride content during challenge with hyposmotic solutions under open circuit (OC) conditions. CH maximally increased following hyposmotic challenge within approximately 5 minutes. The change in CH was smaller under short circuit (SC) conditions or following replacement of chloride in the mucosal solution by gluconate or cyclamate (Cl(-)-freem). When corrected for the osmotically inactive cell volume (30 +/- 2%), delta CH for controls (OC) were greater than predicted for an ideal osmometer. In contrast, delta CH for Cl(-)-freem or SC conditions were similar to that predicted for an ideal osmometer. Na+ and Cl- mucosa-to-serosa fluxes increased following hyposmotic challenge. Chloride fluxes increased maximally within 5 min, then decreased. In contrast, the Na+ flux increased slowly and reached a steady state after approximately 25 min. Under isosmotic conditions, exposure to Cl(-)-freem solutions led to decreases in the transepithelial conductance, Na+ flux, and CH. Chloride permeabilities in the apical and basolateral membranes were detected using the fluorescent intracellular chloride indicator MQAE. The results indicate that during osmotic swelling, the entry of both sodium and chloride is increased. The time courses of these increases differ, suggesting distinct mechanisms for the osmotic regulation of these apical membrane transport processes.
Crowe
W E
WE
Department of Physiology and Biophysics, University of Texas Medical Br., Galveston 77555, USA.
Ehrenfeld
J
J
Brochiero
E
E
Wills
N K
NK
D000818:Animals; D002460:Cell Line; D002463:Cell Membrane Permeability; D016764:Cell Polarity; D048429:Cell Size; D002712:Chlorides / Q000378:metabolism*; D003494:Cyclamates / Q000378:metabolism; D004848:Epithelium / Q000378:metabolism; D005456:Fluorescent Dyes; D005942:Gluconates / Q000378:metabolism; D007424:Intracellular Fluid / Q000378:metabolism; D007686:Kidney Tubules, Distal / Q000166:cytology / Q000378:metabolism*; D009995:Osmosis; D012964:Sodium / Q000378:metabolism*
D002712:Chlorides; D003494:Cyclamates; D005456:Fluorescent Dyes; D005942:Gluconates; D012964:Sodium; C030691:gluconic acid