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Denny, M F; Hare, M F; Atchison, W D
Toxicology and applied pharmacology, 10/1993, Letnik: 122, Številka: 2Journal Article
The effects of the neurotoxic organomercurial methylmercury (MeHg) on intrasynaptosomal polyvalent cation concentrations were examined using fura-2. In the presence of extracellular Ca2+ (Ca2+e), MeHg caused a concentration-dependent, biphasic elevation in the ratio of fluorescence intensity at the emission wavelength of 505 nm following excitation at 340 and 380 nm (340/380 nm ratio). The first phase was independent of Ca2+e and complete within 5 sec. The second phase was dependent upon Ca2+e and was not complete within 6 min. MeHg increased the synaptosomal membrane permeability to Mn2+, suggesting that the second phase was due to influx of Ca2+e. Ruthenium red (20 microM), mitochondrial depolarization (10 mM NaN3 plus 4 micrograms/ml oligomycin), thapsigargin (1 microM), or caffeine (40 mM) did not elevate Ca2+i or alter the response of the synaptosomes to MeHg. Upon closer inspection, we noticed that MeHg simultaneously increased the fluorescence intensity at the excitation wavelengths of 340 and 380 nm and at the Ca(2+)-insensitive excitation wavelength of 360 nm. Pretreatment of synaptosomes with the cell-permeant heavy metal chelator TPEN (50 microM) blocked the MeHg-induced elevations in the 360-nm intensity and the 340/380 nm ratio. TPEN given after MeHg reversed the elevations in the 360-nm intensity. The cell-impermeant heavy metal chelator DTPA (150 microM) had no effect. We conclude that MeHg disrupts polyvalent cation homeostasis by at least two mechanisms. The first involves release of endogenous non-Ca2+ polyvalent cations, while the second is due to increased Ca2+ permeability of the plasma membrane.
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