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    Hg(2+) and Cu(2+) interfere with agonist-mediated Ca(2+) signaling in isolated Mytilus digestive gland cells

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    The effects of mercury and copper on agonist-mediated Ca-signaling were investigated in isolated cells from the marine mussel: Mytilus galloprovincialis Lam., by single cell fluorescence microscopy. In isolated digestive gland cells, short-term exposure (10 min) to both Hg2+, a highly toxic metal and Cu2+, essential metal, in the nano-low mu M range caused a sustained increase in cytosolic [Ca2+]. The effect of mercury on resting [Ca2+] was stronger than that of copper. The Hg-induced elevation in [Ca2+] seemed to be mainly due to an increased influx through Verapamil-sensitive Ca-channels, whereas the effect of Cu2+ was related to a release from thapsigargin-sensitive intracellular stores. Agonists, such as epidermal growth factor (EGF), bradykinin (BK) and ATP, evoked Ca2+ transients in isolated digestive gland cells through different mechanisms similar to those observed in mammalian cells, demonstrating the presence of common pathways of Ca-mediated cell signaling in both invertebrates and vertebrates. The agonist-mediated Ca2+ response was affected by exposure to Hg2+ and Cu2+ in a concentration dependent manner: both metals significantly reduced the amplitude of the Ca2+ spikes elicited by BK and ATP and decreased the percentage of EGF-responsive cells. The effects of Hg2+ and Cu2+ were apparently independent of their different type of interaction with the mechanisms involved in Ca2+ homeostasis. The results clearly demonstrate that, in marine invertebrate cells, short-term exposure to heavy metal concentrations comparable to environmental exposure levels results in alterations of intracellular Ca2+ homeostasis which compromise the cell response to extracellular stimuli involving Ca-mediated signaling. The mechanisms of heavy metal interference with Ca-homeostasis and signaling are discussed. (C) 2000 Elsevier Science B.V. All rights reserved

    Heavy metals and glutathione metabolism in mussel tissues

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    Data are presented on the tissue glutathione content and on the activity of two of the main enzymes involved in GSH metabolism (γ-glutamyl-cysteine synthetase (GCS), GSH transferase (GST)) in the gills and digestive gland of mussels (Mytilus galloprovincialis Lam.) exposed for 1, 4 and 7 days to sublethal concentrations of copper and of mercury, both in the inorganic form of HgCl2 and in the organic form of CH3HgCl. The results indicate that Cu2+ (0.6 μM) and CH3Hg+ (0.2 μM) can affect, although to a different extent, glutathione metabolism in mussel tissues; common traits were a decrease in tissue glutathione content, stimulation of GST activity and inhibition of GSH synthesis rate; on the other hand, inorganic Hg2+ (0.2 μM) did not significantly affect glutathione content and metabolism in mussel tissues. Exposure to copper (0.6 μM) resulted in a decrease in total glutathione content in both gills and the digestive gland that was maximal after the first day, followed by a tendency to recover at longer exposure times. Such a decrease seemed to be mainly related to a stimulation of GST activity, which was transient in gills and persistent in the digestive gland. The tissue glutathione content was also decreased in mussel exposed to methylmercury (0.2 μM); however, such a decrease seemed to be mainly due to inhibition of the GSH synthesis rate and, to a lesser extent, to an increased GST activity. On the contrary, inorganic Hg2+ (0.2 μM) did not significantly affect glutathione metabolism, although mussel exposure to the same concentration of organic and inorganic Hg2+ resulted in similar total metal tissue load in mussel tissues. The role of glutathione as a potential defense against heavy metals is discussed in relation to different routes of metal toxicity and homeostasis in mussel tissues. Copyright (C) 1999 Elsevier Science B.V
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