title: Hg(2+) and Cu(2+) interfere with agonist-mediated Ca(2+) signaling in isolated Mytilus digestive gland cells.
abstract: 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 Hg(2+), a highly toxic metal and Cu(2+), an essential metal, in the nano-low µM range caused a sustained increase in cytosolic [Ca(2+)]. The effect of mercury on resting [Ca(2+)] was stronger than that of copper. The Hg-induced elevation in [Ca(2+)] seemed to be mainly due to an increased influx through Verapamil-sensitive Ca-channels, whereas the effect of Cu(2+) was related to a release from thapsigargin-sensitive intracellular stores. Agonists, such as epidermal growth factor (EGF), bradykinin (BK) and ATP, evoked Ca(2+) 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 Ca(2+) response was affected by exposure to Hg(2+) and Cu(2+) in a concentration dependent manner: both metals significantly reduced the amplitude of the Ca(2+) spikes elicited by BK and ATP and decreased the percentage of EGF-responsive cells. The effects of Hg(2+) and Cu(2+) were apparently independent of their different type of interaction with the mechanisms involved in Ca(2+) 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 Ca(2+) 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.
authors.affiliation: Istituto di Scienze Fisiologiche, Università di Urbino, Campus Universitario-Loc. Crocicchia, 61029, Urbino, Italy
authors.initials: L
authors.lastname: Canesi
authors.initials: C
authors.lastname: Ciacci
authors.initials: G
authors.lastname: Gallo