首页> 外文期刊>The biochemical journal >Membrane retrieval in the guinea-pig neurohypophysis. Isolation and characterization of secretory vesicles and coated microvesicles after radiolabel incorporation in vivo
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Membrane retrieval in the guinea-pig neurohypophysis. Isolation and characterization of secretory vesicles and coated microvesicles after radiolabel incorporation in vivo

机译:豚鼠神经垂体的膜修复。体内放射性标记掺入后分泌囊泡和包被微囊的分离和表征

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pWe have developed small-scale methods for the isolation and biochemical characterization of subcellular fractions from single guinea-pig posterior-pituitary glands. Secretory vesicles and coated microvesicles produced in this way were of similar purity to those isolated from large amounts of tissue by conventional ultracentrifugation. [35S]Cysteine injected into the hypothalamus was found in the soluble contents of secretory vesicles isolated from the neural lobes 24 h later. High-pressure liquid-chromatographic analysis revealed that the radiolabel was incorporated into the expected neurosecretory products (oxytocin, vasopressin and neurophysin) and also into a biosynthetic intermediate in the vasopressin system. The membranes of secretory vesicles were labelled with [3H]choline 24 h after its hypothalamic injection. Little or no [3H]choline could be demonstrated in coated microvesicles at this time, although these structures were labelled 5 days after injection. Stimulating hormone secretion by chronic dehydration produced a significant fall in [3H]choline content of the secretory-vesicle membranes without any transfer of label into coated microvesicles, suggesting that coated microvesicles are not involved in membrane retrieval in the neurohypophysis./p
机译:>我们已经开发了小规模的方法,用于从单个豚鼠后垂体腺分离和生化表征亚细胞级分。以这种方式产生的分泌性囊泡和包被的微囊泡与通过常规超速离心从大量组织中分离出的那些具有相似的纯度。在下丘脑中注射的[35S]半胱氨酸在24小时后从神经叶分离出的分泌性囊泡的可溶性成分中被发现。高压液相色谱分析表明,放射性标记已被掺入预期的神经分泌产物(催产素,血管加压素和神经元)中,并且也被掺入到血管加压素系统的生物合成中间体中。下丘脑注射后24小时,用[3H]胆碱标记分泌性囊泡的膜。尽管在注射后5天标记了这些结构,但此时在包被的微囊泡中几乎没有[3H]胆碱被证实。慢性脱水刺激激素分泌会导致分泌囊泡膜的[3H]胆碱含量显着下降,而没有任何标记转移到被覆微囊泡中,这表明被覆微囊泡不参与神经垂体的膜修复。

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