ROLE OF CYTOPLASMIC FREE CA2+ IN SECRETION

提出了一种方法，从而可以测量细胞质中的 离子钙（Ca-1）浓度和激素分泌率 相同的细胞群。 Ca-i用发光蛋白测定 水母发光蛋白，其通过2种方法之一加载到细胞中：低渗 冲击或刮擦加载。 我建议确定合并的方法 最多的水母发光蛋白，同时对细胞的损伤最小， 法 我建议测量变化的幅度和时间进程 在Ca-i和由许多促分泌素引起的激素分泌速率中， 以确定肌醇中cAMP变化如何影响这些变化 三磷酸盐代谢以及雌二醇和三碘甲状腺原氨酸激素。 初步结果表明，quin-2可以缓冲 所以我建议确定 quin-2的掺入影响促分泌素对细胞内Ca-i的作用， 水母发光蛋白负载细胞。 最后，我建议使用水母发光蛋白来测试 三磷酸肌醇对大鼠离体心肌细胞钙释放的影响 透化细胞的细胞内储存，并确定如何释放 可能被cAMP和钙调素改变。

A method is presented whereby it is possible to measure the cytoplasmic concentration of ionized calcium (Ca-i) and rates of hormone secretion from the same population of cells. Ca-i is measured with the photoprotein aequorin which is loaded into the cells by 1 of 2 methods: hypoosmotic shock or scrape loading. I propose to determine the method to incorporate the most aequorin while producing the least damage to the cells with this method. I propose to measure the magnitude and time course of the changes in Ca-i and rates of hormone secretion evoked by a number of secretagogues, to determine how these changes are influenced by cAMP changes in inositol triphosphate metabolism, and the hormones estradiol and triidothyronine. Preliminary results suggest that quin-2 may buffer the effects of the secretagogues on Ca-i and hormone so I propose to determine if the incorporation of quin-2 affects the effect of secretagogues on Ca-i of aequorin loaded cells. Finally I propose to use aequorin to test the effects of inisotol triiphosphate on the release of calcium from intracellular stores of permeabilized cells and determine how the release may be altered by cAMP and calmodulin.