A Method to Isolate Rabbit Arterial Organelles in a Pure and Functional State as Determined by a Marker Enzyme and an Organelle Specific Enzyme
Yourick, Jeffrey J.
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Atherosclerosis is a disease which causes lipids to accumulate within the intima of arteries. Lipid accumulation leads to the development of morphologic lesions in the artery. The metabolic state of the mitochondria is also altered in this disease process. In studying the pathways of cholesterol biosynthesis and changes in the metabolic rates of the arterial cell, it is necessary to isolate the specific organelles responsible for each function. The goal of this study was to develop a method for isolating mitochondria and microsomes from rabbit aortas. Methods have been developed for organelle separation in rats and swine, but little work has been done with rabbits. The method was developed by maximizing organelle purity according to a marker enzyme, succinate dehydrogenase. Homogenization time; tissue handling (stripping, layering, and mincing); temperature; centrifugation times; and buffers were considered. Mitochondrial function was tested by an ADP-ATP translocase assay. The results provide a method for rabbit arterial organelle separation.
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