The influence of calcium transients on intracellular pH in cortical neurons in primary culture.

The objective of this study was to assess the influence of Ca2+ influx on intracellular pH (pHi) of neocortical neurons in primary culture. Neurons were exposed to glutamate (100-500 microM) or KCl (50 mM), and pHi was recorded with microspectrofluorometric techniques. Additional experiments were carried out in which calcium influx was triggered by ionomycin (2 microM) or the calcium ionophore 4-Br-A23187 (2 microM). Glutamate exposure either caused no, or only a small decrease in pHi (delta pH approximately 0.06 units). When a decrease was observed, a rebound rise in pHi above control was observed upon termination of glutamate exposure. In about 20% of the cells, the acidification was more pronounced (delta pH approximately 0.20 units), but all these cells had high control pHi values, and showed gradual acidification. Exposure of cells to 50 mM KCl consistently increased pHi. Since this increase was similar in the presence and nominal absence of HCO3-, it probably did not reflect influx of HCO3- via a Na(+)-HCO3- symporter. Furthermore, since it occurred in the absence of external Ca2+ (or a measurable rise in Cai2+) it seemed independent of Ca2+ influx. It is tentatively concluded that the rise in pHi was due to reduced passive influx of H+ along the electrochemical gradient, which is reduced by depolarization. In Ca(2+)-containing solutions, depolarization led to a rebound increase in pHi above control. This, and the rebound found after glutamate transients, may reflect Ca(2+)-triggered phosphorylation and upregulation of the Na+/H+ antiporter which extrudes H+ from the cell.(ABSTRACT TRUNCATED AT 250 WORDS)