alexa Oxidative injury triggers autophagy in astrocytes: the role of endogenous zinc.


Journal of Neuroinfectious Diseases

Author(s): Lee SJ, Cho KS, Koh JY, Lee SJ, Cho KS, Koh JY

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Abstract We have recently demonstrated that the accumulation of labile zinc in lysosomes during oxidative stress causes lysosomal membrane permeabilization (LMP) in cultured hippocampal neurons. Since autophagy involves fusion of autophagic vacuoles (AVs) with lysosomes, zinc accumulation may start in AVs. In the present study, we examined the role of endogenous zinc in H2O2-induced autophagy and cell death in mouse astrocyte cultures. Live-cell confocal imaging of astrocytes transfected with GFP-LC3 revealed that the number of AVs positive for LC3 (microtubule-associated protein 1 light chain 3) increased following exposure to H2O2 or ferrous chloride (FeCl2). Staining of RFP-LC3-transfected astrocytes with FluoZin-3 indicated that the levels of labile zinc increased in AVs as well as in the cytosol and nuclei. The majority of AVs were double-stained with LysoTracker, indicating that they were fused with lysosomes. Chelation of zinc with tetrakis [2-pyridylmethyl]ethylenediamine (TPEN) decreased the number of AVs in H2O2-treated astrocytes, whereas exposure to zinc increased their number, suggesting that dysregulation of zinc homeostasis is mechanistically linked to autophagy. Unexpectedly, inhibition of autophagy blocked the rise in labile zinc levels. Astrocytic death induced by H2O2) was ccompanied by LMP. Autophagy inhibitors (3-methyladenine, bafilomycin-1) or TPEN attenuated LMP and cell death in astrocytes. These results support the possibility that endogenous zinc plays a key role in autophagy under oxidative stress in astrocytes, and suggest that autophagy is a necessary preceding event for LMP and cell death in oxidative injury. This article was published in Glia and referenced in Journal of Neuroinfectious Diseases

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