Effects of pH on intracellular calcium levels in isolated cochlear outer hair cells of guinea pigs.

Abstract

Effects of intracellular pH (pHi) on intracellular Ca2+ (Cai2+) in the outer hair cell (OHC) were investigated using fluorescence ratio imaging microscopy. Cai2+ and pHi were determined with fluorescence indicators fura-2 and 2,7-bis(2-carboxyethyl)- 5,6-carboxyfluorescein, respectively. Intracellular alkalinization from the basal pHi of 7.22 +/- 0.03 to 7.82 +/- 0.09 (n = 7) induced by 25 mM NH4+ caused a rise in Cai2+ from 102 +/- 15 to 195 +/- 28 nM (n = 8). The elevation of Cai2+ was inhibited by removing external Ca2+ or by 50 microM nifedipine. On the other hand, the cytosolic acidification produced by the removal of NH4+ (delta pHi = 0.53 +/- 0.06, n = 7) and the admission of 5% CO2 (delta pHi = 0.72 +/- 0.05, n = 4) elicited a slight reduction in Cai2+. The depolarization of the membrane potential by exposure to 100 mM K+ induced an increase in Cai2+ that was susceptible to nifedipine, indicating the presence of voltage-sensitive Ca2+ channels. The elevation in Cai2+ induced by 100 mM K(+)-containing solution was increased by 138 +/- 6% (n = 4) by external alkalinization to pH 8.4 and was decreased by 87 +/- 3% (n = 4) by external acidification to pH 6.4. These results demonstrate that both internal and external alkalinization of OHCs facilitate the influx of Ca2+ through the Ca2+ channel, leading to a rise of Cai2+. Thus pH, extra- and intracellular, may modulate the OHC motility by regulating the Cai2+.