Advisor: Kenneth A. Barbee, Ph.D.
This study details the investigation of key proteins in the nitric oxide production pathway in microvessels. Nitric oxide is an essential molecule in the prevention of disease states like atherosclerosis, hypertension, clotting disorders, and autoimmune disease. While large vessel endothelial cells are regularly studied for nitric oxide pathway proteins, little data has been produced on the distribution of these proteins in microvessels. Previous cell culture models have demonstrated heterogeneous calcium responses as part of the nitric oxide production pathway when microvessel cells are stimulated with ATP. Here we present the characterization of spatial heterogeneity of the purinergic receptor P2Y2 in rat adrenal medulla capillary cells, as well as the distributions of the microdomain caveolae (identiﬁed by caveolin-1) and endothelial nitric oxide synthase, in vivo.
Furthermore, we present a program used to analyze immunohistochemical images to quantify the expressio n of the proteins of interest within the endothelium. Immunohistochemical staining of these proteins was performed and analyzed for ﬂuorescence intensity and distribution within the tissue’s endothelium. The Kolmogorov-Smirnov Test was employed to verify statistical signiﬁcance of the experimental data from controls. The study reveals populations of endothelial cells that express clearly bimodal distributions of the purinergic receptor, indicating spatial heterogeneity. Caveolin-1 and endothelial nitric oxide synthase distributions are uniform and colocalized, as previous research indicates in other tissues.
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