Effects of ethanol on intracellular-free Ca2+ concentration in cultured rat aortic smooth muscle cells were examined by digital imaging fluorescence microscopy using the Ca2+ fluorescence indicator, fura-2. Ethanol induced dose-dependent decrements in cytosolic-free Ca2+ concentration at 45 mM and 90 mM, which was consistent with previously reported observations of relaxation in intact rat aortic tissues. However, ethanol at high pharmacological concentrations (e.g., 450 mM) failed to induce any further inhibition in cytosolic-free Ca2+ concentration. Our results suggest that the vasodilator effects of ethanol, observed on intact blood vessels, may result in part from an interference with the availability of Ca2+ for excitation-contraction coupling in vascular smooth muscle cells.
Effects of extracellular magnesium ions ([Mg2+]o) on intracellular free Mg2+ ([Mg2+]i) and its subcellular distribution in single fission yeast cells, Schizosaccharomyces pombe, were studied with digital-imaging microscopy and an Mg2+ fluorescent probe (mag-fura-2). Using 0.44 mM [Mg2+]o, [Mg2+]i in yeast cells was 0.91 +/- 0.08 mM. Elevation of [Mg2+]o to 1.97 mM induced rapid (within 5 min) increments in [Mg2+]i (2.18 +/- 0.11 mM). Lowering [Mg2+]o to 0.06 mM, however, exerted no significant effects on [Mg2+]i (0.93 +/- 0.14 mM), at least for periods of up to 30 min. Irrespective of the [Mg2+]o used, the subcellular distribution of [Mg2+]i remained heterogeneous, i.e. where the sub-plasma membrane region > cytoplasm > nucleus. [Mg2+] in all three subcellular compartments increased significantly, two- to threefold, concomitant with [Mg2+] when placed in 1.97 mM [Mg2+]o. We conclude that [Mg2+]i in fission yeast is maintained at a physiologic level when [Mg2+]o is low, but intracellular free Mg2+ rapidly rises when [Mg2+]o is elevated. Like most eukaryotic cells, yeast may have a Mg2+ transport system(s) which functions to maintain gradients of Mg2+ from the outside to inside the cell and among its subcellular compartments.
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