pH dependence and compartmentalization of zinc transported across plasma membrane of rat cortical neurons.

In this study, Zn(2+) transport in rat cortical neurons was characterized by successfully combining radioactive tracer experiments with spectrofluorometry and fluorescence microscopy. Cortical neurons showed a time-dependent and saturable transport of (65)Zn(2+) with an apparent affinity of 15-20 microM. (65)Zn(2+) transport was pH dependent and was decreased by extracellular acidification and increased by intracellular acidification. Compartmentalization of newly transported Zn(2+) was assessed with the Zn(2+)-selective fluorescent dye zinquin. Resting cortical neurons showed uniform punctate labeling that was found in cell processes and the soma, suggesting extrasynaptic compartmentalization of Zn(2+). Depletion of intracellular Zn(2+) with the membrane-permeant chelator N,N,N',N'-tetrakis(2-pyridylmethyl)-ethylenediamine (TPEN) resulted in the complete loss of punctate zinquin labeling. After Zn(2+) depletion, punctate zinquin labeling was rapidly restored when cells were placed in 30 microM Zn(2+), pH 7.4. However, rapid restoration of punctate zinquin labeling was not observed when cells were placed in 30 microM Zn(2+), pH 6.0. These data were confirmed in parallel (65)Zn(2+) transport experiments.