Coupling of cell volume and membrane potential changes to fluid secretion in a model of renal cysts

Coupling of cell volume and membrane potential changes to fluid secretion in a model of renal cysts. Renal tubular epithelia ordinarily absorb NaCl and water, although recent evidence indicates that renal cysts secrete fluid. We have utilized the experimental advantages offered by cultured cysts, formed in a collagen matrix by propagating Madin-Darby canine kidney cells, to investigate the mechanisms involved in fluid secretion by this renal epithelium. The rate of fluid transport (adduced from changes in cavity volume), cell volume and changes in membrane potential were measured simultaneously in isolated cysts. Under basal conditions, cysts absorbed fluid (-0.83 ± 0.34 × 10−6 ml/min/cm2 cavity surface area, N = 23). AVP and IBMX changed the direction of net fluid transport to secretion (4.24 ± 0.49 × 10−6 ml/min/cm2. Cell volume initially fell 7.4 ± 0.5% and remained stable thereafter as secretion continued. Membrane electrical potential (bis-oxonol epifluorescence) hyperpolarized in 13 cysts and depolarized in 6, the mean change was 1.9 ± 3.1%. Fluid secretion was abolished by 0.1mM ouabain. Secretion was not affected by 0.1mM DIDS and cell pH (bis-carboxyethyl-carboxyfluorescein epifluorescence) was not altered by the induction of secretion, suggesting that secretion is not dependent on Cl-HCO3 exchange. Barium, in the presence of AVP and IBMX, depolarized the cell membrane potential (bis-oxonol fluorescence increased 22.3 ± 0.03%), reversed secretion to absorption (from 3.21 ± 0.93 to -1.52 ± 0.61 × 10−6 ml/min/cm2), and increased cell volume 2.7 ± 0.5%. Bumetanide (100 µM) reduced fluid secretion from 4.49 ± 1.23 to -0.75 ± 0.55 × 10−6 ml/min/cm2, further reduced cell volume 4.4 ± 1.2% and hyperpolarized the membranes (bis-oxonol fluorescence fell 24.3 ± 5.0%). In the absence of AVP and IBMX bumetanide had no effect on fluid transport, cell volume or membrane potentials. We conclude that AVP reversed the direction of fluid transport in these cultured renal epithelial cysts from absorption to secretion by stimulating a coordinated interaction of basolateral and apical K, Cl and Na transport mechanisms