CO 2/HCO 3−-withdrawal from the bath medium of hippocampal slices: biphasic effect on intracellular pH and bioelectric activity of CA3-neurons

Abstract

Many studies analyzing interactions of pH and bioelectric activity focus on changes of the extracellular pH, whereas data concerning central neuronal excitability and intracellular pH (pH i) are rare. Here, we report on the spontaneous bioelectric activity and epileptiform activity of CA3-neurons during a procedure which changed pH i. As monitored in BCECF-AM loaded cells, the change from a CO 2/HCO 3 −-buffered to a HEPES-buffered medium (CO 2/HCO 3 −-withdrawal, hereafter termed W) was associated with a transient intracellular alkalosis (ΔpH=0.2±0.04) which preceded a sustained intracellular acidosis (ΔpH=0.4±0.04). Coinciding with this W-induced biphasic shift of pH i a biphasic alteration of spontaneous bioelectric activity was recorded: as a rule, an up to 30 min lasting increase (excitatory phase) preceded a typical sustained suppression (inhibitory phase). This biphasic action was also observed using various in vitro-epilepsy-models (bicuculline, penicillin, caffeine): epileptiform discharges were completely suppressed after an initial increase in frequency. This modulation of bioelectric activity was unlikely due to alterations of the postsynaptic GABA-system as hyperpolarizing GABA A- and GABA B-responses of CA3-neurons were hardly affected. In the majority of the neurons, the initial increase of spontaneous bioelectric activity (excitatory phase) culminated in transient burst periods lasting 5–30 min. These transient burst periods were blocked by NMDA- or AMPA-antagonists: dl-2-amino-5-phosphonovalerate (APV, 50 μM) or 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 50 μM). The calcium-antagonist verapamil (50 μM) reduced amplitudes of depolarizations and duration of the transient burst periods. The results suggest that the biphasic alteration of pH i modulates the susceptibility of glutamate receptors and voltage-gated calcium-channels, which leads to respective changes of bioelectric activity.