Effects of transmembrane gradients of bicarbonate and ammonium on junctional and nonjunctional membrane conductances in BHK cells.
Weak acids are efficient blockers of gap-junctional conductance. It is generally accepted that intracellular acidification produced by weak acids fully accounts for the gap-junctional uncoupling. Protonation of the cytoplasmic portions of the channel-forming protein connexin is thought to lead to the conformational changes switching the channel from the open into the closed state. If this is the only mechanism of the weak-acid induced uncoupling, then the correlation between junctional conductance (Gj) and intracellular pH (pHin) should not depend on the means of intracellular acidification. We compared the responses of junctional conductance in BHK cells measured in double whole-cell experiments to the applied transmembrane concentration gradients of bicarbonate or ammonium. These treatments were to lower pHin in a predictable way according to the equations: pHin = pHout -lg[[HCO3]out/HCO3-]in) or pHin = PHout - lg[[NH4+]in[NH4+out), respectively. We found that the behavior of Gj depended on the substance used. At a 500-fold bicarbonate gradient (calculated pHin approximately 4.8) the cells remained coupled, while a 100- or 10-fold gradient of ammonium imposing pHin approximately 6.1 produced fast uncoupling. The responses of junctional conductance were often accompanied or preceded by changes of non-junctional membrane conductance. We suggest that the mechanisms of the weak acid/base-induced channel gating may contain an additional "lipophilic" component due to the presence of the non-dissociated form of the acid/base in cell membrane.
Dunina-Barkovskaya AYA
Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Russia. dunina@genebee.msu.su
Bujurina
I M
IM
Pivovarov
V S
VS
Frolov
V A
VA
D000818:Animals; D001639:Bicarbonates / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D006224:Cricetinae; D004553:Electric Conductivity; D004594:Electrophysiology; D017629:Gap Junctions / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D000644:Quaternary Ammonium Compounds / Q000378:metabolism
D001639:Bicarbonates; D000644:Quaternary Ammonium Compounds