Alteration in Intracellular Zn(2+) Homeostasis as a Result of TRPM2 Channel Activation Contributes to ROS-Induced Hippocampal Neuronal Death

Transient receptor potential melastatin-related 2 (TRPM2) channel, a molecular sensor for reactive oxygen species (ROS), plays an important role in cognitive dysfunction associated with post-ischemia brain damage thought to result from ROS-induced TRPM2-dependent neuronal death during reperfusion. Emerging evidence further suggests that an alteration in the Zn(2+) homeostasis is critical in ROS-induced TRPM2-dependent neuronal death. Here we applied genetic and pharmacological interventions to define the role of TRPM2 channel in ROS-induced neuronal death and explore the mechanisms contributing in the alteration in intracellular Zn(2+) homeostasis in mouse hippocampal neurons. Exposure of neurons to 30–300 μM H(2)O(2) for 2–24 h caused concentration/duration-dependent neuronal death, which was significantly suppressed, but not completely prevented, by TRPM2-knockout (TRPM2-KO) and pharmacological inhibition of the TRPM2 channel. H(2)O(2)-induced neuronal death was also attenuated by treatment with TPEN acting as a Zn(2+) selective chelator. Single cell imaging demonstrated that H(2)O(2) evoked a prominent increase in the intracellular Zn(2+) concentration, which was completely prevented by TPEN as well as TRPM2-KO and inhibition of the TRPM2 channel. Furthermore, H(2)O(2) induced lysosomal Zn(2+) release and lysosomal dysfunction, and subsequent mitochondrial Zn(2+) accumulation that provokes mitochondrial dysfunction and ROS generation. These H(2)O(2)-induced lysosomal/mitochondrial effects were prevented by TRPM2-KO or TPEN. Taken together, our results provide evidence to show that a dynamic alteration in the intracellular Zn(2+) homeostasis as a result of activation of the TRPM2 channel contributes to ROS-induced hippocampal neuronal death.