Mechanisms of neuroprotection against oxidative stress in the anoxia tolerant turtle Trachemys scripta

Abstract

The detrimental effects of oxidative stress caused by the accumulation of Reactive Oxygen Species (ROS) have been acknowledged as major factors in aging, senescence, and several neurodegenerative conditions including Parkinson's Disease and stroke (ischemia/reperfusion). Mammalian models are extremely susceptible to these stresses and the restoration of oxygen after anoxia causes oxidative damage; however some organisms including the freshwater turtle Trachemys scripta can withstand such conditions without any apparent pathology. T. scripta thus provides us with a model in which we can investigate physiological mechanisms of neuroprotection in anoxia and reoxygenation without the damaging effects that come with oxidative stress.The ability of the turtle to survive these stressors is thought to be related to its high innate levels of antioxidants and its capacity to suppress ROS production. One potential antioxidant mechanism is the Methionine Sulfoxide Reductase System (Msr) that catalyzes the reduction of oxidized Methionine (Met) residues. Methionine residues are easily oxidized amino acids and oxidized Met is associated with decreased protein activity. Msr may also restore function to damaged proteins and it has been shown to scavenge ROS and thus ameliorate cellular damage.The present study investigated the role and regulation of MsrA in turtle primary neuronal cell cultures exposed to 4 hours of anoxia and 4 hours of anoxia followed by 4 hours of reoxygenation. Anoxia and anoxia/reoxygenation increase MsrA mRNA and protein levels, which suggests a protective role under these conditions. MsrA expression was then altered with target specific siRNA; we found that the knock down of MsrA expression resulted in decreased cell viability. As the factors that control MsrA expression in this model are still unknown, we also induced FOXO3a expression by transfecting a FOXO3a plasmid into the cells, as it has been shown to regulate MsrA levels in other animal models. The induction of FOXO3a protected neurons against cell death. Furthermore, treatment of cells with the green tea extract Epigallocatechin gallate (EGCG), which has antioxidant properties, results in increased expression of FOXO3a when the cells are exposed to oxidative stress. Together these results suggest that MsrA may play an important role in protection against anoxia and oxidative stress in the turtle brain, and may prove to be a target to treat diseases of ischemia and oxidative stress.Support or Funding InformationNIH‐NIA Project # 1R15AG033374‐01,FAU FoundationThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.