Cell Physiology Core

细胞生理学核心

• 批准号: 10625323
• 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/10625323
• 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; 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; absorption; 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 项目摘要 Pannexin 1 (Panx1) 形成质膜离子通道，释放重要的细胞间信号传导 病理和生理条件下的分子，特别是 ATP 和 UTP；渠道还支持 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）。在此 PPG 应用中，采用新型小鼠模型来确定 Panx1- 的贡献 介导 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、优化和标准化关键信息，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。其中包括：（1）Panx1通道功能分析 通过电生理学和染料摄取测定对原代细胞进行检测； (2)小说的电生理表征 Pannexin 通道的小分子调节剂。同样，核心也在开发中占据主动 可以为整个研究计划提供明显的技术优势的创新工具和方法。这 包括通过 CRISPR/Cas9 生成新型 Panx1 变体和基因编辑小鼠品系 技术。在执行这些通常对整个研究计划至关重要的集中目标时，核心 认识到数据质量和再现性的至关重要性。对于垂直和水平 该 PPG 中提出的集成，该核心位于蜂窝和系统分析级别之间的接口 Panx1 功能。