title: Activation of functionally protective K(+) channels by methylmercury in rat alveolar macrophages.
abstract: Methylmercury (MeHg) is generally known as a neurotoxic heavy metal while its effect on alveolar macrophages is still rarely studied. In this paper, we attempted to use whole cell and cell-attached patch-clamp recording technique and fura-2 fluorescence measurement to elucidate the effects of MeHg on rat alveolar macrophages. The results showed that extracellular application of MeHg induced a transient outward current I(O)(MeHg), 10-20 s in duration, 100-1000 pA in amplitude at -40 mV associated with a marked increase in conductance. The reversal potential depended distinctly on the external K(+) concentration. Removal of external Ca(2+) as well as bath applied verapamil caused a depression of I(O)(MeHg), and intracellular dialysis with 5 mM EGTA completely abolished I(O)(MeHg). Heparin (5 mg/ml) applied by intracellular dialysis greatly accelerated a run-down of I(O)(MeHg) induced by pressure ejection of MeHg. K(+) channel blockers such as quinine, and 4-aminopyridine especially low concentrations of dequalinium and apamin, but not tetraethylammonium inhibited I(O)(MeHg). Cell-attached single-channel recordings with the pipette solution containing 145 mM KCl revealed that the activation of single-channel currents with a conductance of 12 pS could be induced by application of MeHg outside the patch. Since MeHg increased [Ca(2+)](i), in a concentration-dependent manner which was partially blocked by either verapamil or Ca(2+)-free medium containing 1 mM EGTA, it is concluded that MeHg activates a Ca(2+)-dependent K(+) conductance by an increase of [Ca(2+)](i) through an influx from outside the cells as well as mobilization from intracellular store. A possibility that this membrane hyperpolarizing K(+) current may exhibit a functioning modulator in response to the harmful cytotoxic increase in [Ca(2+)](i) caused by MeHg was tested. Accordingly, this working hypothesis is verified by an increase of MeHg-induced cytotoxicity of cultured rat alveolar macrophages through a blockade of this Ca(2+)-activated K(+) channel by dequalinium.
authors.affiliation: Institute of Toxicology, College of Medicine, National Taiwan University, Taipei, Taiwan.
authors.forename: T C
authors.initials: TC
authors.lastname: Kuo
authors.forename: S Y
authors.initials: SY
authors.lastname: Lin-Shiau