SALIVARY MUCIN SYNTHESIS, PROCESSING, AND SECRETION

这项提议的一个关键要素是为IN开发一个现场系统 一种粘蛋白刺激-分泌-合成循环的深度分析 小鼠的颌下腺。收集到的信息将应用于 对控制合成和加工的机制的理解 唾液粘蛋白。刺激-分泌-合成系统将是 标准化，以便学到的东西可以作为未来的参考 唾液腺功能障碍的研究，如口干、流涎和 慢性肥大。 粘蛋白的量、合成率和降解率 在周期的每个阶段利用放射免疫测定法进行定量 粘蛋白多聚体的免疫沉淀。此外，还有一个协议是 所描述的将提供糖基化事件的准确时间。 翻译与寡糖之间可能的调控相互作用 组装将在衣霉素的帮助下进行研究。的影响 不同剂量不同促分泌剂对小鼠皮肤的影响 将比较刺激-分泌-合成周期。 一种允许评估粘蛋白含量的过程 在刺激分泌之前，将使用个体的腺体。流动 唾液粘蛋白和蛋白质含量以及新生粘蛋白的比率 将获得腺体的多肽和粘蛋白多聚体含量。 最后，粘蛋白低聚糖进行加工的可能性 将对分泌的步骤进行调查。将使用一种协议， 提供来自同一个体的粘蛋白的碳水化合物分析 分泌物分泌前后。

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.