We tested the ability of methylmercury (MeHg) to block calcium channel current in cultures of neonatal cerebellar granule cells using whole-cell patch clamp techniques and Ba(2+) as charge carrier. Low micromolar concentrations of MeHg (0.25-1 microM) reduced the amplitude of whole cell Ba(2+) current in a concentration- and time-dependent fashion; however, this effect was not voltage-dependent and the current-voltage relationship was not altered. Increasing the stimulation frequency hastened the onset and increased the magnitude of block at both 0.25 and 0.5 microM MeHg but not at 1 microM. In the absence of stimulation, all concentrations of MeHg were able to decrease current amplitude. The ability of several Ca(2+) channel antagonists (omega-conotoxin GVIA, omega-conotoxin MVIIC, omega-agatoxin IVA, calcicludine, and nimodipine) to alter the MeHg-induced effect was tested in an effort to determine if MeHg targets a specific subtype of Ca(2+) channel. Each of the antagonists tested was able to decrease a portion of whole cell Ba(2+) current under control conditions. However, none were able to attenuate the MeHg-induced block of whole cell Ba(2+) current, suggesting either that the mechanism of MeHg-induced block involves sites other than those influenced specifically by Ca(2+) channel antagonists or that MeHg was able to "outcompete" these toxins for their binding sites. These results show that acute exposure to submicromolar concentrations of MeHg can block Ba(2+) currents carried through multiple Ca(2+) channel subtypes in primary cultures of cerebellar granule cells. However, it is unlikely that the presence of a specific Ca(2+) channel subtype is able to render granule cells more susceptible to the neurotoxicologic actions of MeHg.
Copyright 2000 Academic Press.