Induction of apoptosis in human T-cells by methyl mercury: Temporal relationship between mitochondrial dysfunction and loss of reductive reserve

Abstract

The objective of our study was to define the mechanism by which MeHgCl induces human T-cell apoptosis. We asked the question: does mercury disrupt the ΔΨ(m) and induce a mitochondrial permeability transition state? Using two fluorescent reagents, JC-1 and DiOC6(3), we demonstrated that MeHgCl exposure resulted in a decrease in the ΔΨ(m). Since a decline in ΔΨ(m) can disturb the ph(i), we employed SNARF-1 to assess ph(i); results indicate that mercury treatment reduced the ph(i) from 7.0 to 6.5. Consistent with these observations, we noted that uncoupled electron transfer reactions generated ROS, while cardiolipin, a mitochondrial phospholipid, was oxidized. In concert with the biochemical changes, there was a decrease in overall dimension of the mitochondria of mercury-treated cells and a loss in cristae architecture. The toxicant also depleted the thiol reserves of the cell and promoted translocation of cytochrome c from the mitochondria to the cytosol. Furthermore, when T cells were thiol-depleted, there was increased susceptibility to MeHgCl-induced apoptosis. Finally, we established a temporal relationship between the decline in ΔΨ(m), generation of ROS, and depletion of thiol reserves. The earliest detectable event was at the level of the mitochondrion; in the presence of MeHgCl there was a profound reduction in mitochondrial ΔΨ(m) and a decline in GSH lewis within 1 h. Subsequently, a further decrease in thiol reserves was linked to the generation of ROS. We propose that the target organelle for MeHgCl is the mitochondrion and that induction of oxidative stress leads to activation of death-signaling pathways.