ROLE OF CYTOPLASMIC FREE CA2+ IN SECRETION

细胞质游离 CA2 在分泌中的作用

• 批准号: 3236775
• 负责人: KENNETH W SNOWDOWNE
• 金额: $ 11.52万
• 依托单位: UNIVERSITY OF THE PACIFIC-SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-02-01 至 1989-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3236775
• 关键词: calcium; cyclic AMP; cytoplasm; estradiol; hormone regulation /control mechanism; inositol; ionic strengths; membrane permeability; secretion; triiodothyronine

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.

提出了一种方法，可以测量细胞质 电离钙的浓度（CA-1）和激素分泌的速率 相同的细胞群。 用光蛋白测量CA-I 通过2种方法中的1种将其加载到细胞中的Aquorin：低湿 冲击或刮擦负载。 我建议确定合并的方法 在此对细胞造成最小损害时，最燃烧的是 方法。 我建议测量变化的幅度和时间过程 在CA-1和许多促销员引起的激素分泌率中， 确定这些变化如何受肌营的营地变化的影响 三磷酸代谢，以及激素雌二醇和三甲基氨酸。 初步结果表明，Quin-2可能会缓冲 关于CA-I和激素的促分子，我建议确定是否确定 Quin-2的掺入会影响秘密的影响对CA-I的影响 Aquorin负载细胞。 最后，我建议使用aequorin测试 三磷酸二霉菌对钙从释放钙释放的影响 透化细胞的细胞内存储，并确定如何释放 营地和钙调蛋白可能会改变。