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The main goal of the research is to characterize the antioxidant activity associated with normal prion protein in both mouse neuroblastoma and bovine fibroblast cell lines.
This research will help us determine the normal physiological function of prion protein, which has not been completely elucidated at this point. The first objective is to determine the amount protein expressed in both normal and prion knocked-down mouse neuroblastoma and bovine fibroblast cell lines. The second objective is to characterize the antioxidant activity in both control and prion knock-down cell lines. Our third objective is to measure and compare biomarkers indicative of oxidative stress in both the control cell lines and their knock-down counterparts.
The normal physiological function of prion protein has not been completely elucidated at this time. Normal prion protein may exhibit neuroprotective functions. The purpose of this study is to determine if prion protein is associated with antioxidant activity or decreased levels of oxidative damage.
To accomplish the first objective, control and knock-down cell cultures will be grown and harvested. An ELISA assay will then be used to determine the amount of prion expression in each sample. For the second objective, antioxidant activity will be characterized using a commercially available spectrophotometric assay kit to measure superoxide dismutase activity defined as the ability to catalyze the conversion of superoxide radicals into molecular oxygen and hydrogen peroxide. Additionally, measurement of the ratio of reduced glutathione to oxidized glutathione will provide an indication of the antioxidant status in our samples. For the third objective, biomarkers of lipid peroxidation and DNA damage, indicators of oxidative stress will be measured using ELISA and GC-MS analytical techniques.