Free intracellular Ca2+ concentration ([Ca2+]i, Ca2+ currents, and excitatory amino acid (EAA) currents were studied in spinal neurons cultured in low (4.5 mM) and high (25 mM) extracellular potassium. When challenged with lethal concentrations of N-methyl-D-aspartate (NMDA) or kainate, neurons cultured in 25 mM K+ exhibited markedly attenuated Ca2+ currents and [Ca2+]i responses, and survived the EAA challenge more readily than controls. Surprisingly, NMDA and Kainate currents remained comparable between neurons grown in high- and low K+. The disparity between the observed [Ca2+]i increases and EAA currents suggests that chronic depolarization induces a fundamental alteration in intracellular Ca2+ handling. This phenomenon may provide clues for the development of neuroprotective strategies against excitotoxin excess.