Bicarbonate-chloride exchange in gill plasma membranes of blue crab

The uptake of chloride was studied in gill plasma membrane vesicles from low-salinity-adapted blue crabs (Callinectes sapidus). Cl- uptake was not Na+ dependent. However, when a HCO-3 gradient (in greater than out) was imposed across the membrane, a transient overshoot of about 2.5-fold was produced. Approximately 90% of the Cl- uptake reflected entry into the osmotically active intravesicular space. Cl- itself, nitrate, hydroxyl, and sulfite could substitute for HCO-3. The HCO-3/Cl- exchange process appeared to saturate at higher concentrations of either HCO-3 or Cl-. The apparent Km for Cl- was 15 mM. HCO-3-dependent Cl- uptake was significantly inhibited by 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid and thiocyanate but not by amiloride, furosemide, or ouabain. Alterations in membrane potential had no effect on Cl- uptake. Addition of Cl- or HCO-3 to the external medium also accelerated efflux of 36Cl- and H14CO-3 from preloaded vesicles. These results indicate that the uptake of Cl- by the crab gill plasma membrane is a carrier-mediated Na+-independent anion exchange process.