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 By replacing the failing and outdated parts with new state-of-the-art microscope components, this upgrade will significantly increase the capability and throughput of our integrated confocal-TIRF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.

抽象的 这是一份用于升级集成旋转磁盘的行政补充申请 PI 实验室中的共焦和全内反射荧光 (TIRF) 显微镜系统。这 进行母体中提出的所有活细胞成像实验需要显微镜系统 该项目的重点是确定 Nir2 蛋白调节 PIP2 稳态的机制 受体诱导的 Ca2+ 信号传导过程中的内质网 (ER)-质膜 (PM) 连接。 这种集成的共焦 TIRF 显微镜系统是专门为监测动态变化而设置的。 受体期间相同细胞中 PM PIP2 水平和蛋白质易位至 ER-PM 连接 刺激。我们研究所的成像核心设施没有这样的系统。最近， 这个 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。迫切需要升级这种集成显微镜系统，以防止破坏 由于系统故障而导致父项目。此外，此升级还将集成新的 技术融入显微镜系统，以满足我们未来五到十年的成像需求。这 UT 西南大学生理学系将通过提供资金来支持这一升级 平衡超出预算，并为升级的显微镜系统提供空间。经过 这次升级用新的最先进的显微镜组件替换了故障和过时的部件 将显着提高我们集成共焦 TIRF 显微镜的功能和通量 系统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其影响力。