ISOLATION OF A GOLGI APPARATUS-RICH FRACTION FROM RAT LIVER : I. Method and Morphology |
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Authors: | D. James Morr , R. L. Hamilton, H. H. Mollenhauer, R. W. Mahley, W. P. Cunningham, R. D. Cheetham, V. S. Lequire |
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Affiliation: | From the Departments of Botany and Plant Pathology and Biology, Purdue University, Lafayette, Indiana 47907; Departments of Anatomy and Pathology, Vanderbilt University Medical School, Nashville, Tennessee 37203; Charles F. Kettering Research Laboratory, Yellow Springs, Ohio 45387; and the Department of Genetics and Cell Biology, University of Minnesota, St. Paul, Minnesota 55101 |
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Abstract: | Golgi apparatus were released without fixatives from rat hepatocytes by gentle homogenization, concentrated by differential centrifugation, and purified by sucrose gradient centrifugation. Examination of sections of purified fractions by electron microscopy showed fields of morphologically intact units of Golgi apparatus consisting of stacks of parallel flattened cisternae, secretory vesicles, and small vesicular profiles. Negative staining of unfixed pellets revealed a complex network of anastomotic tubules continuous with platelike structures and secretory vesicles. These structures corresponded, respectively, to the small vesicular profiles and parallel flattened cisternae with attached secretory vesicles of sectioned material. Small fragments of granular endoplasmic reticulum were often closely associated with the peripheral tubules, suggesting sites of continuity in intact hepatocytes. |
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