SALIVARY MUCIN SYNTHESIS, PROCESSING, AND SECRETION

该提案的一个关键要素是开发一个现场系统， 对一种粘蛋白的刺激-分泌-合成循环的深入分析， 老鼠的下颌下腺 收集到的信息将用于 为了理解控制合成和加工的机制， 唾液粘蛋白 刺激-分泌-合成系统 标准化，以便所学到的可以作为未来的参考 唾液腺功能障碍如口干、流涎和 慢性肿大 粘蛋白的量、合成速率和降解速率将 在周期的每个阶段通过利用放射免疫测定进行定量， 粘蛋白多核糖体的免疫沉淀。 此外，一个协议是 所描述的方法将提供糖基化事件的准确定时。 翻译和寡糖之间可能的控制相互作用 将在衣霉素的帮助下研究组装。 的影响 不同剂量的促分泌素 刺激-分泌-合成周期进行比较。 本发明提供了一种程序，其允许评估免疫球蛋白的粘蛋白含量。 将使用在分泌刺激之前的个体的腺体。 流 唾液中粘蛋白和蛋白质含量以及新生粘蛋白 将获得腺体的多肽和粘蛋白多核糖体内容物。 最后，粘蛋白低聚糖进行加工的可能性 将研究分泌步骤。 将使用一个协议， 提供来自同一个体的粘蛋白的碳水化合物分析 分泌前后。

A key element of this proposal is to develop an in situ system for an in depth analysis of the stimulus-secretion-synthesis cycle of a mucin from the submandibular glands of mice. The information gathered will be applied toward understanding mechanisms for control of synthesis and processing of salivary mucin. The stimulus-secretion-synthesis system will be standardized so that what is learned may serve as reference for future studies of salivary gland dysfunction such as xerostomia, sialorrhea and chronic enlargement. The amounts, rates of synthesis and rates of degradation of the mucin will be quantitated at each phase of the cycle by utilizing radioimmunoassay and immunoprecipitation of mucin polysomes. In addition, a protocol is described that will provide accurate timing of the glycosylation events. Possible control interactions between translation and oligosaccharide assembly will be investigated with the help of tunicamycin. The effects of various doses of different secretagogues on the stimulus-secretion-synthesis cycle will be compared. A procedure which allows for an assessment of the mucin content of an individual's glands prior to secretory stimulation will be used. Flow rates, mucin and protein contents of saliva, as well as mucin nascent polypeptides and mucin polysome contents of glands will be obtained. Finally, the possibility that mucin oligosaccharides undergo a processing step at secretion will be investigated. A protocol will be used that provides for carbohydrate analyses of the mucin from the same individual before and after secretion.