Na+-dependent H+ and Cl- transport in in vitro frog gastric mucosa

Na+ dependency of H+ and Cl- transport in resting and stimulated gastric mucosae was investigated. We measured transepithelial potential difference (PD), short-circuit current (Isc), conductance, H+ secretion (JH), and unidirectional fluxes of 36Cl- (muscosal (m) to serosal (s), JClm leads to s and JCLs leads to m) during Isc conditions in Ussing-type chambers. Resting tissues: Na+-free serosal solution (choline replacement), but not mucosal, Ringer solution caused PD and Isc to decrease to zero with a time course identical to that observed with Cl--free solutions. Conductance also decreased by more than 50%. Isc = JCLs leads to m--JCLm leads to 8 = JClnet during control ([Na+] = 105 mM), and Na+-free conditions. When [Na+] = 50 mM (choline replacement) Isc (=JClnet) was reduced by approximately 25%; when K+ replaced Na+, Isc (=JClnet) was reduced by approximately 65%. The dependence of Isc on [Na+] (choline replacement) was sigmoidal indicating that there may be two sites at which Na+ activates Cl- secretion. Plots of 1/Isc vs. 1/[Cl-] with different [NA+] indicated, that the affinity of the Cl- "carrier" may be affected by [Na+]. stimulated tissues: Na+-free (choline replacement) solutions caused PD, Isc, and JClnet all to decrease to zero but sometimes Na+-containing mucosal solution caused JH to increase back to 50% of control, whereas Isc increased by only 16%. Na+-free effects were not reversed by 1 mM dibutyryl cAMP plus 0.1 mM isobutyl methylxanthine in the serosal solution. As [Na+] was increased, Isc (sigmoidal) and JH (monotonic) both increased. K+ replacement of Na+ caused a larger decrease in Isc than when choline was used; JH remained constant if [Na+] greater than or equal to mM. We have proposed that the energy-dependent step for active Cl- transport occurs at the serosal membrane in a Na+-coupled process. Cl- that accompanies JH during open-circuit conditions may utilize a separate system.