pH regulation in the vertebrate central nervous system: microelectrode studies in the brain stem of the lamprey

Abstract

Studies of intracellular pH (pHi) in nervous tissue are summarized and recent investigation of intracellular and extracellular pH (pHo) in the isolated brain stem of the lamprey is reviewed. In the lamprey, pHi regulation was studied in single reticulospinal neurons using double-barrel ion-selective microelectrodes (ISMs). In nominally [Formula: see text]-free HEPES-buffered media, acute acid loading was followed by a spontaneous recovery of pHi requiring 10–20 min and was associated with a prolonged rise in intracellular Na+. The recovery of pHi was blocked by 1–2 mM amiloride. Amiloride also caused a small rise in pHo. Substitution of external Na+ caused a slow intracellular acidification and extracellular alkalinization. Return of external Na+ reversed these effects. Transition from HEPES to [Formula: see text]-buffered media increased the rate of acid extrusion during recovery of pHi. Recovery in [Formula: see text]-buffered media was inhibited by 4,4′-diisothio-cyanostilbene-2,2′-disulfonic acid and was slowed after exposure to Cl−-free media. Following inhibition of acid extrusion by amiloride, transition to [Formula: see text] media restored pHi recovery. These data indicate that lamprey neurons recover from acute acid loads by both Na+–H+ exchange and an independent [Formula: see text]-dependent mechanism. Evidence for [Formula: see text]-dependent acid extrusion in other vertebrate cells and the protocols of pHi studies using ISMs are discussed.