Depolarization-induced increase in synaptosomal membrane calcium monitored by chlorotetracycline fluorescence

Chlorotetracycline (CT) was used as a fluorescent probe for membrane calcium with intact synaptosomes. The net increase in fluorescence intensity at 520 nm, which is a measure of membrane-bound Ca, increases with increasing Ca, saturating in the millimolar range. Membrane Ca can also be detected in the absence of added external Ca. Potassium-induced depolarization of synaptosomes leads to an increase in membrane Ca, reaching a new steady-state value within 5 min. Neither opiates nor phenytoin affected synaptosomal membrane Ca. Relatively high concentrations of chlorotetracycline increased depolarization-induced uptake of 45Ca into synaptosomes. The data suggest that the Ca-CT complex binds to synaptic plasma membranes, and that depolarization-induced Ca influx results in increased Ca binding to the internal surface of the plasma membrane and/or other internal membranes.