The primary goal for this fellowship is to acquire advanced training in cell biology and transport physiology by examining the effects of endotoxin administration on hepatocellular transport function. Sinusoidal and canalicular hepatocyte membrane vesicles will be prepared from endotoxin-treated rats and control animals. The kinetics of 3H- taurocholate uptake will be determined using an ultra-rapid filtration technique. Endotoxin-induced alterations in membrane vesicular Na+/K+ ATPase activity will then be assessed using a coupled kinetic enzymatic assay. Western immunoblotting will also be performed to measure endotoxin-induced changes in protein expression of the Na+/K+ ATPase, and alterations in membrane fluidity will be determined using fluorescence polarization. In accomplishing this goal, I will expand my knowledge of cellular transport physiology and protein/membrane biology and acquire the skills and techniques required for further investigation in these fields.
