Chloride distribution in the proximal convoluted tubule of Necturus kidney.
To assess the mechanism(s) by which intraluminal chloride concentration is raised above equilibrium values, intracellular Cl- activity (alpha iCl) was studied in the proximal tubule of Necturus kidney. Paired measurements of cell membrane PD (VBL) and Cl-selective electrode PD (VBLCl) were performed in single tubules, during reversible shifts of peritubular or luminal fluid composition. Steady-state alpha iCl was estimated at 14.6 +/- 0.6 mmol/liter, a figure substantially higher than that predicted for passive distribution. To determine the site of the uphill Cl- transport into the cell, an inhibitor of anion transport (SITS) was added to the perfusion fluid. Introduction of SITS in peritubular perfusate decreased alpha iCl, whereas addition of the drug in luminal fluid slightly increased alpha iCl; both results are consistent with basolateral membrane uphill Cl- transport from interstitium to the cell. TMA+ for Na+ substitutions in either luminal or peritubular perfusate had no effect on alpha iCl. Removal of bicarbonate from peritubular fluid, at constant pH (a situation increasing HCO3- outflux), resulted in an increase of alpha iCl, presumably related to enhanced Cl- cell influx: we infer that Cl- is exchanged against HCO3- at the basolateral membrane. The following mechanism is suggested to account for the rise in luminal Cl- concentration above equilibrium values: intracellular CO2 hydration gives rise to cell HCO3- concentrations above equilibrium. The passive exit of HCO3- at the basolateral membrane energizes an uphill entry of Cl- into the cell. The resulting increase of alpha iCl, above equilibrium, generates downhill Cl- diffusion from cell to lumen. As a result, luminal Cl- concentration also increases.
D000818:Animals; D001639:Bicarbonates / Q000494:pharmacology; D001692:Biological Transport / Q000187:drug effects; D002462:Cell Membrane / Q000502:physiology; D002712:Chlorides / Q000378:metabolism*; D004566:Electrodes; D007687:Kidney Tubules, Proximal / Q000187:drug effects / Q000378:metabolism*; D007700:Kinetics; D008564:Membrane Potentials; D009337:Necturus; D010477:Perfusion; D011188:Potassium / Q000494:pharmacology; D012964:Sodium / Q000494:pharmacology
D001639:Bicarbonates; D002712:Chlorides; D012964:Sodium; D011188:Potassium