FUNCTIONAL DEVELOPMENT OF THE SALIVARY GLANDS

这个项目的总体目标是描述 潜在机制或途径的出生后发展 唾液腺分泌液体和电解质。我们 建议使用一种制备分散细胞的方法 1日龄、7日龄、14日龄和21日龄大鼠下颌下腺的观察 在新的授权期内，这条道路的几个方面。在……里面 成熟的腺体，现在已知它涉及离子的激活 运输系统，这导致了跨腺泡运动 钠和氯，以及由此产生的渗透梯度，水。其他 有证据表明，唾液腺的刺激会增强，因为 在其他各种组织中，膜的周转 磷脂，导致形成起作用的产物 手机信使。尽管两者之间的确切关系 这两个组成部分的分泌性反应并不完全 明白了，有证据表明他们之间有关联 通过细胞内钙离子的变化。人们对此知之甚少 这个复杂的分泌物的各种成分的外观 唾液腺发育过程中的反应。我们建议 因此，研究以下内容：1)跨膜通量 在同位素示踪剂的帮助下，单价离子(Na，K，C1)。 这些离子的吸收和外流将在不存在的情况下测量。 在促分泌剂、转运抑制剂和 特定的通道阻滞剂。动力学性质(即时间 课程、费率、半最高值等)和外部离子 将调查每种助焊剂的需求；2)刺激- 诱导磷脂周转。形成的时间进程 和肌醇磷酸盐、二酰甘油和 花生四烯酸的色谱学研究 放射性标记分子的分离；3) 上述2)中确定的产物对钙离子的释放 细胞内储存与细胞外钙离子动员 环境。同位素示踪剂(45Ca)和 为此，将使用Fura-2的荧光分光光度法。这个 这两种钙离子运动对单价离子的影响 还将通过测量离子(同位素)来研究运输。 钙离子通道存在时，上述1)中所示的通量 阻滞剂和将其捕获到细胞存储器中的物质(即TMB- 8)。这些研究的结果将表明该复合体是否 商店反应在出生后发育的所有阶段都起作用 以及它的所有组成元素是否在 在此期间的同一时间。

The overall objective of this project is to characterize the postnatal development of the mechanism or pathway underlying the secretion of fluid and electrolytes in salivary glands. We propose to use a preparation of dispersed cells from the submandibular gland of 1, 7, 14 and 21 day old rats to investigate in the new grant period several aspects of this pathway. In mature glands, it is now known to involve the activation of ion transport systems, which results in the transacinar movement of Na and Cl and, by the resulting osmotic gradient, of water. Other evidence indicates that stimulation of salivary glands enhances, as in a variety of other tissues, the turnover of membrane phospholipids, leading to the formation of products which act as cellular messengers. Although the exact relationship between these two components of the secretory response is not fully understood, there is suggestive evidence that they are linked through changes in cell calcium. Little is known about the appearance of the various components of this complex secretory response during salivary gland development. We propose to investigate, therefore, the following: 1) transmembrane fluxes of monovalent ions (Na, K, C1) with the aid of isotopic tracers. Uptake and efflux of these ions will be measured in the absence and in the presence of secretagogues, transport inhibitors and specific channel blockers. The kinetic properties (i.e., time courses, rates, half maximal values, etc.) and the external ionic requirements of each flux will be investigated; 2) stimulation- induced turnover of phospholipids. The time course of formation and breakdown of inositol phosphates, diacylglycerol and arachidonic acid will be investigated by chromatographic separation of radiolabeled molecules; 3) the effects of the products identified in 2) above on the release of Ca++ from intracellular stores and on Ca++ mobilization from the external environment. Both isotopic tracers (45 Ca) and the spectrofluorimetry of fura-2 will be used for this purpose. The effects of these two types of Ca++ movement on monovalent ion transport will also be investigated by measuring the ion (isotope) fluxes indicated in 1) above in the presence of Ca++ channel blockers and of substances which trap it in cell stores (i.e., TMB- 8). The results of these studies will indicate whether the complex stores response operates at all stages of postnatal development and whether all its component elements become functional at the same time during this period.