Cell Physiology Core

细胞生理学核心

• 批准号: 10407612
• 负责人: BIMAL N. DESAI
• 金额: $ 31.48万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10407612
• 关键词: Antibodies; Apoptotic; Biological Assay; Blood Cells; Blood Vessels; CRISPR/Cas technology; Cardiometabolic Disease; Cell Line; Cell membrane; Cell physiology; Cells; Cues; Development; Dyes; Electrophysiology (science); Equipment; Evaluation; Flow Cytometry; Fluorescent Dyes; Generations; Genes; Goals; Gold; Human; Individual; Inflammation; Ion Channel; Measurement; Measures; Mediating; Methods; Mission; Mus; Pathologic; Physiological; Physiology; Reagent; Reproducibility; Research; Services; Signal Transduction; Signaling Molecule; Standardization; System; TO-PRO-3; Techniques; Training; Variant; data quality; design; innovation; mouse model; novel; patch clamp; programs; response; small molecule; tool; uptake; vascular inflammation

CORE C PROJECT SUMMARY Pannexin 1 (Panx1) forms plasma membrane ion channels that release important intercellular signalling molecules, specifically ATP and UTP, under pathological and physiological conditions; the channels also support uptake of various fluorescent dyes that have traditionally been used to mark apoptotic cells (e.g., YO-PRO-1, TO-PRO-3). In this PPG application, novel mouse models are employed to ascertain contributions of Panx1- mediated ATP/UTP release, (and other metabolite release, when needed) to various aspects of cardiometabolic diseases, and to understand corresponding mechanisms of Panx1 channel activation. Cell Physiology Core centralizes the functional analysis of Pannexin channels by absorbing, optimizing and standardizing the key assays of universal significance to all four projects. These include: (1) Analysis of Panx1 channel function in primary cells by electrophysiology and dye uptake assays and; (2) Electrophysiological characterization of novel small molecule modulators of Pannexin channels. Similarly, the core also takes the initiative in the development of innovative tools and methods that can provide a clear technical edge to the entire research program. This includes generation of novel Panx1 variants and gene-edited mouse lines, generated by CRISPR/Cas9 technology. In executing these centralized goals, often of salience to the entire research program, the core recognizes the paramount importance of data quality and reproducibility. For the vertical and horizontal integration proposed in this PPG, this core sits at the interface between cellular and systems levels of analysis of Panx1 function.

核心C项目总结 PAnnexin 1(Panx1)形成质膜离子通道，释放重要的细胞间信号 分子，特别是ATP和UTP，在病理和生理条件下；通道还支持 摄取传统上用于标记凋亡细胞的各种荧光染料(例如，YO-PRO-1， To-Pro-3)。在这个PPG应用中，使用了新的小鼠模型来确定Panx1- 介导的ATP/UTP释放(以及需要时的其他代谢物释放)到心脏代谢的各个方面 疾病，并了解Panx1通道激活的相应机制。细胞生理学核心 通过吸收、优化和标准化关键字，集中进行PAnnexin通道的功能分析 对所有四个项目都具有普遍意义的分析。主要包括：(1)对PANX1通道功能的分析 原代细胞的电生理学和染料摄取分析；(2)新型的电生理表征 PAnnexin通道的小分子调节剂。同样，核心也掌握了发展的主动权 创新的工具和方法，可以为整个研究计划提供明显的技术优势。这 包括产生新的Panx1变体和由CRISPR/Cas9产生的经过基因编辑的小鼠系 技术在执行这些集中的目标时，通常是整个研究计划的重点，核心 认识到数据质量和再现性的最高重要性。对于垂直和水平 在此PPG中提出的集成，此核心位于蜂窝和系统分析级别之间的接口 Panx1函数。