CHROMAFFIN GRANULE MOVEMENT NEAR THE PLASMA MEMBRANE

质膜附近的嗜铬颗粒运动

• 批准号: 2642251
• 负责人: LAURA M JOHNS
• 金额: $ 2.09万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-16 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2642251
• 关键词: atrial natriuretic peptide; chimeric proteins; chromaffin cells; exocytosis; fluorescence microscopy; granule; green fluorescent proteins; intracellular transport; secretory protein; tissue /cell culture

DESCRIPTION (Adapted from applicant's abstract): Although much is known about exocytosis in chromaffin cells, little is known about secretory granule movement close to the plasma membrane preceding the final fusion event. A key aspect of the proposal is our ability to visualize chromaffin granules in real time. A fusion protein of atrial natriuretic peptide and green fluorescent protein that is expressed by transient transfection is sorted to chromaffin granules. Using the optical technique of total internal reflection, we can visualize fluorescent chromaffin granules in living cells that are localized within 1-2 granule diameters of the plasma membrane. I hypothesize that movement and placement of chromaffin granules close to the plasma membrane are highly regulated processes that help define the availability of granules to undergo exocytosis. The focus of this proposal is to determine whether cofactors and proteins known to influence the final secretory response act at least in part by controlling these prefusion events. My specific aims are the following: (1) to determine the location and movement of chromaffin granules close to the plasma membrane under basal and secretory conditions, (2) to determine the relationship between the known effects of ATP and Ca2+ on granule priming before secretion and granule movement adjacent to the plasma membrane and (3) to determine the role of SNAREs and Rab3a, specific proteins of the secretory pathway in granule movement and placement adjacent to the plasma membrane.

描述（改编自申请人的摘要）：虽然知道很多 关于嗜铬细胞的胞吐作用，对分泌型的知之甚少。 在最后融合之前，颗粒运动靠近质膜 活动 该提案的一个关键方面是我们可视化嗜铬细胞的能力 颗粒在真实的时间。 一种心房利钠肽与 通过瞬时转染表达的绿色荧光蛋白， 分类为嗜铬颗粒。 利用全反射的光学技术 内反射，我们可以看到荧光嗜铬颗粒， 位于血浆1-2个颗粒直径内的活细胞 膜的 我假设嗜铬颗粒的移动和位置 靠近质膜的是高度调节的过程， 进行胞吐作用的颗粒的可用性。 的重点 一项建议是确定已知的辅助因子和蛋白质是否影响 最终分泌反应至少部分地通过控制这些 融合前事件。 我的具体目标是：（1）确定 嗜铬颗粒靠近质膜的位置和运动 在基础和分泌条件下，（2）确定 ATP和Ca 2+对颗粒启动的已知作用之间的关系， 分泌和颗粒运动邻近质膜和（3）， 确定SNARE和Rab 3a的作用，分泌的特异性蛋白质， 颗粒运动的途径和邻近质膜的位置。