Homeostatic Regulation of PIP2-Calcium Signaling

PIP2-钙信号传导的稳态调节

• 批准号: 10797897
• 负责人: JEN LIOU
• 金额: $ 23.68万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-15 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10797897
• 关键词: Administrative Supplement; Budgets; Calcium Signaling; Cell membrane; Cells; Core Facility; Endoplasmic Reticulum; Equilibrium; Failure; Funding; Grant; Homeostasis; Image; Investments; Measurement; Microscope; Monitor; Phosphatidylinositol 4,5-Diphosphate; Physiology; Process; Protein translocation; Proteins; Regulation; Resolution; Signal Transduction; System; experimental study; fluorescence microscope; live cell imaging; new technology; parent project; prevent; quantitative imaging; receptor

Abstract This is an administrative supplement application for funding to upgrade an integrated spinning-disk confocal and total internal reflection fluorescence (TIRF) microscope system in the PI’s lab. This microscope system is required for carrying out all live-cell imaging experiments proposed in the parent project that is focused on defining mechanisms regulating PIP2 homeostasis by Nir2 protein at endoplasmic reticulum (ER)-plasma membrane (PM) junctions during receptor-induced Ca2+ signaling. This integrated confocal-TIRF microscope system is uniquely set up for monitoring dynamic changes of PM PIP2 levels and translocation of proteins to ER-PM junctions in the same cells during receptor stimulation. Such a system is not available at the Imaging Core Facility of our institute. Recently, several components of this microscope system that was set up in 2009 are losing functionality at a fast pace. An upgrade of this integrated microscope system is urgently needed to prevent disruptions of the parent project due to failure of the system. In addition, this upgrade will enable integration of new technology into the microscope system to meet our imaging needs for the next five to ten years. The Physiology Department at UT Southwestern will support this upgrade by providing funds to cover the balance exceeding the budget, and by providing space for the upgraded microscope system. By replacing the failing and outdated parts with new state-of-the-art microscope components, this upgrade will significantly increase the capability and throughput of our integrated confocal-TIRF microscope system to obtain high-quality quantitative imaging results for years to come. This is an investment that will greatly facilitate the progress and enhance the impact of the parent project.

摘要 这是一个行政补充申请资金，以升级一个集成的旋转磁盘 共聚焦和全内反射荧光（TIRF）显微镜系统在PI的实验室。这 需要一个显微镜系统来进行母公司中提出的所有活细胞成像实验 该项目的重点是确定Nir 2蛋白调节PIP 2稳态的机制， 内质网（ER）-质膜（PM）连接在受体诱导的Ca 2+信号。 这种集成的共焦TIRF显微镜系统是唯一的设置，用于监测动态变化， PM PIP 2水平和蛋白质在受体介导期间在相同细胞中转运到ER-PM连接处 刺激.我们研究所的成像核心设施没有这样的系统。最近， 2009年建立的显微镜系统的几个组件正在快速失去功能 步伐。迫切需要对这种集成显微镜系统进行升级，以防止 由于系统故障而导致的父项目。此外，此次升级将使新的 我们希望将这项技术应用到显微镜系统中，以满足我们未来五到十年的成像需求。的 UT西南生理学系将通过提供资金来支持这一升级， 这意味着我们可以节省超过预算的资金，并为升级后的显微镜系统提供空间。通过 用新的最先进的显微镜组件替换故障和过时的部件， 将显着提高我们集成的共焦TIRF显微镜的能力和吞吐量 该系统可在未来几年内获得高质量的定量成像结果。这是一项投资， 将极大地促进母项目的进展并增强母项目的影响。