Analysis of Ca2+ homeostasis in neurons dissociated from rat nucleus basalis.

Whole-cell patch-clamp recordings of calcium currents (ICa) and fura-2 microfluorimetric measurements of intracellular free Ca2+ concentration ([Ca2+]i) were made simultaneously in neurons acutely dissociated from rat nucleus basalis. Depolarization activated ICa and caused an increase in [Ca2+]i. The relationship between total Ca2+ influx and the increase in [Ca2+]i was studied. After repolarization, [Ca2+]i recovered to control values within a few seconds. A mathematical model was constructed to simulate the mechanisms underlying [Ca2+]i regulation; the parameters were (1) the rate of Ca2+ influx, (2) the rate of the [Ca2+]i increase by the Ca2+ influx, and (3) the rate of Ca2+ clearance from cytosol due to extrusion across the plasma membrane and sequestration into calcium storing organelles. After an appropriate evaluation of parameter values from the experimental results, the model mimicked the processes of [Ca2+]i increase and recovery. The experimental results and simulations suggest that (1) neurons possess a large Ca2+ buffering capacity, (2) systems for Ca2+ clearance are activated by the Ca2+ influx in a saturable manner, (3) the rate of Ca2+ clearance is relatively small compared to the rate of Ca2+ influx evoked by depolarizations, and (4) the shoulder in the [Ca2+]i recovery phase is due to the asymptote of the Ca2+ clearance rate.