Intracellular Zinc Trafficking in Human Umbilical Vein Endothelial Cells During Noxious Injury

Maintaining and regulating intracellular zinc is a fundamental process critical to cell function. Hemorrhagic snake venom and other noxious injuries cause the reduction of tissue integrity, production of reactive oxygen species. This project will investigate intracellular zinc mobilization during Crotalus atrox (western diamondback) venom stimulation. Disrupted intracellular zinc homeostasis that maintains high intracellular zinc can trigger mitochondrial dysfunction, increase oxidative stress, and cell death. Although more details are emerging, it is unclear if zinc is mobilized during cellular injury and if venom induced cellular injury alters the expression of metal binding proteins such as metallothioneins (MT) and/or metal transcription factor 1 (MTF1). This project will use advanced fluorescent microscopy to improve our understanding of zinc trafficking by examining intracellular cellular zinc movement with the fluorescent zinc probe, Fluozin-3AM. In addition, we utilized SDS-PAGE and Western Blotting techniques to monitor the protein expression of MT and MTF1 in after Crotalus atrox venom stimulation