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Fractionation of the rat ventral prostate with respect to isolation and exocytosis of the prostatic secretion protein
Authors:Tapio Haaparanta  Åke Pousette  Bertil Högberg  Jan-Åke Gustafsson  Hans Glaumann
Institution:1. Department of Pathology, Karolinska Institute, Huddinge University Hospital, S-141 86 Huddinge, Sweden;2. Medical Nutrition, Karolinska Institute, Huddinge University Hospital, S-141 86 Huddinge, Sweden
Abstract:The ventral prostrate was fractionated into one mitochondrial and three microsomal fractions. The different fractions were characterized morphologically and chemically. An interesting finding was that upon homogenization the endoplasmic reticulum membranes often turned ‘inside-out’ giving rise to microsomes with ribosomes attached to the inside of the vesicles. The secretion of the protatic secretion was studied by means of isotopic pulse labeling using radioactive leucine. Peak radioactivity in the secretory fluid was obtained at 2 h after injection with a relativity rapid fall. The radioactivity in the secretory fluid displayed a continuous increase up to 8 h followed by a plateau. When prostatic secretion was purified from secretory fluid and microsomes using a Con A-Sepharose column it showed a typical precursor-product relationship with an early peak at 60 min in microsomal prosatatic secretion protein followed by a peak in secretory fluid at 4 h. Vinblastine blocked the release of labeled secretion protein into the secretory fluid, a phenomenon characteristic for secretory proteins which are exocytosed by means of fusion between secretory granules and the plasma membrane. Following intravenous injection of 3H]estramustine, accumulation was seen in the secretory fluid. Some estramustine probably binds to newly synthesized protatic secretion protein and follows the same route of intracellular transport and extracellular discharge as does prostatic secretion protein.
Keywords:Exocytosis  Secretion protein  Estramustine  Pulse labeling  (Rat ventral prostate)
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