Hypotonic stress-induced release of KHCO3 in fused renal epitheloid (MDCK) cells.
Mechanisms of cell volume regulation induced by the reduction of the osmolality of the Ringer solution by one-third were studied in fused Madin-Darby canine kidney (MDCK) cells. Intracellular HCO3-, K+ and Cl- concentrations [ion]i in parallel with cell membrane potential (PD), cell membrane conductance (Gm) and conductances of individual ions (Gmion) were evaluated with microelectrode techniques. Fused cells regulate their cell volume by about 50%. Gm increased from 0.43 +/- 0.03 mS/cm2 in isotonic Ringer solution to 4.3 +/-0.3 mS/cm2 in the steady state phase of cell swelling. GmCl was 0.31 +/- 0.03 mS/cm2 in isotonic Ringer solution and thus was the dominant individual ion conductance. In the initial phase of cell swelling GmK increased transiently 64-fold to 0.32 +/- 0.03 mS/cm2, and consequently PD hyperpolarized. At peak hyperpolarization GmCl transiently decreased by 15%. Cell swelling increased GmCl 11-fold and GmHCO3 28-fold to 0.95 +/- 0.1 mS/cm2 in the steady state phase of cell swelling. In this phase GmCl and GmHCO3 were dominating, whereas GmK was only slightly increased compared to isotonic conditions. The hyperpolarization of PD was paralleled by cytoplasmic acidification. At peak acidification [HCO3-]i decreased by 6.4 mmol/kg H2O. Cl- extrusion was not detectable in the initial phase of cell swelling. In isotonic Ringer solution [K+]i was 125 +/- 5 mmol/kg H2O. During the initial phase of cell swelling 23 +/- 5 mmol/kg H2O K+ was extruded, indicating that yet unknown anions participated in cell volume regulation in this phase of cell swelling. In the steady state phase of cell swelling [pH]i was normalized by replenishing [HCO3-]i, whereas Cl- was extruded. We conclude that fused renal epitheloid cells acutely release KHCO3 in response to hypotonicity, but then regain pH homeostasis in the steady state phase of cell swelling.
Kersting
U
U
Department of Physiology, University of Würzburg, Germany.
Wojnowski
L
L
Steigner
W
W
Oberleithner
H
H
D000818:Animals; D001639:Bicarbonates / Q000378:metabolism*; D002459:Cell Fusion; D002460:Cell Line; D002712:Chlorides / Q000378:metabolism; D004285:Dogs; D004594:Electrophysiology; D015622:Epithelioid Cells / Q000502:physiology*; D007038:Hypotonic Solutions / Q000494:pharmacology*; D007668:Kidney / Q000502:physiology*; D008564:Membrane Potentials; D008839:Microelectrodes; D011188:Potassium / Q000378:metabolism*; D017680:Potassium Compounds
D001639:Bicarbonates; D002712:Chlorides; D007038:Hypotonic Solutions; D017680:Potassium Compounds; C026329:potassium bicarbonate; D011188:Potassium