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Small alterations in the level of extracellular H +  can profoundly alter neuronal activity throughout the nervous system. In this study, self-referencing H + -selective microelectrodes were used to examine extracellular H +  fluxes from individual astrocytes. Activation of astrocytes cultured from mouse hippocampus and rat cortex with extracellular ATP produced a pronounced increase in extracellular H +  flux. The ATP-elicited increase in H +  flux appeared to be independent of bicarbonate transport, as ATP increased H +  flux regardless of whether the primary extracellular pH buffer was 26 mM bicarbonate or 1 mM HEPES, and persisted when atmospheric levels of CO 2  were replaced by oxygen. Adenosine failed to elicit any change in extracellular H +  fluxes, and ATP-mediated increases in H +  flux were inhibited by the P2 inhibitors suramin and PPADS suggesting direct activation of ATP receptors. Extracellular ATP also induced an intracellular rise in calcium in cultured astrocytes, and ATP-induced rises in both calcium and H +  efflux were significantly attenuated when calcium re-loading into the endoplasmic reticulum was inhibited by thapsigargin. Replacement of extracellular sodium with choline did not significantly reduce the size of the ATP-induced increases in H +  flux, and the increases in H +  flux were not significantly affected by addition of EIPA, suggesting little involvement of Na + /H +  exchangers in ATP-elicited increases in H +  flux. Given the high sensitivity of voltage-sensitive calcium channels on neurons to small changes in levels of free H + , we hypothesize that the ATP-mediated extrusion of H +  from astrocytes may play a key role in regulating signaling at synapses within the nervous system.

Extracellular ATP-Induced Alterations in Extracellular H+ Fluxes From Cultured Cortical and Hippocampal Astrocytes