Effect of glutamate and extracellular calcium on uptake of inorganic lead (Pb 2+) in immortalized mouse hypothalamic GT1–7 neuronal cells

Abstract

We have previously shown that although glutamate alone has no effects on viability of mouse hypothalamic GT1–7 cells, it clearly enhances Pb 2+-induced cytotoxicity. It is likely that Pb 2+ must enter cells to exert most of its toxic effects. Pb 2+ is known to substitute for Ca 2+ in many cellular processes. Therefore, we studied the uptake mechanisms of Pb 2+ into GT1–7 neuronal cells with a special focus on the role of extracellular calcium (Ca 2+), voltage-sensitive calcium channels (VSCCs) and glutamate. Basal uptake of Pb 2+ (1 μM or 10 μM), i.e. without any external stimulus, clearly increased in nominally Ca 2+-free buffer and was partially abolished by 13 mM Ca 2+ when compared to uptake in the presence of a physiological concentration of extracellular Ca 2+ (1.3 mM). Depolarization by 25 mM K +, or antagonists of VSCCs, verapamil (10 μM) or flunarizine (10 μM) had no clear effect on basal Pb 2+ uptake. Glutamate (1 mM) increased Pb 2+ uptake, but only when cells were treated with 1 μM Pb 2+ in the presence of 1.3 mM Ca 2+. Our data suggest that Pb 2+ competes for the same cellular uptake pathways with Ca 2+, although not via VSCCs. In addition, enhancement of Pb 2+-induced neurotoxicity by glutamate may be due to increased neuronal uptake of Pb 2+.