New Chemical Tools for Exploring Cellular Physiology

探索细胞生理学的新化学工具

• 批准号: 9981758
• 负责人: Evan Walker Miller
• 金额: $ 22.09万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-22 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9981758
• 关键词: Area; Biophysics; Cardiac Myocytes; Cell Cycle; Cell membrane; Cell physiology; Cells; Chemicals; Chemistry; Disease; Electrodes; Electron Microscopy; Electron Transport; Generations; Goals; Health; Human; Image; Ions; Length; Life; Measurement; Membrane Potentials; Microscopy; Molecular; Monitor; Neurons; Neurosciences; Organelles; Organic Chemistry; Physiology; Research; Resolution; Role; Specific qualifier value; Synthesis Chemistry; System; Techniques; Time; base; design; insight; migration; patch clamp; programs; reconstruction; sensor; small molecule; temporal measurement; tool; voltage

Abstract New Chemical Tools for Visualizing Cellular Physiology The cell membrane is the only organelle shared by all varieties of cellular life. Unequal distribution of ions and chemical species across the plasma membrane results in the generation of an electrochemical potential, and rapid changes in this membrane potential, or voltage, drive the unique physiology of excitable cells like neurons and cardiomyocytes. However, all cells, even non-excitable cells, possess a membrane potential, and mounting evidence supports a role for membrane potential in controlling fundamental cellular physiology—for example, cell cycle, migration, proliferation, and differentiation—in non-excitable cells. Despite the central role of membrane potential to the cellular physiology of both excitable and non-excitable cells, our understanding of membrane potential in these systems remains incomplete, due in large part to a lack of tools for studying cellular physiology with high spatial and temporal resolution. Measurements of membrane potential rely on highly invasive, low throughput direct voltage recording through electrodes (patch clamping) or by indirectly monitoring the down-stream effects of membrane potential via imaging (Ca2+ imaging). We propose to use the power of synthetic organic chemistry to design fluorescent voltage sensors to probe membrane potential dynamics in neurons and cardiomyocytes, in addition to non-excitable cells. In a complementary approach, we are developing small molecule-based activity integrators that integrate Ca2+ transients over time to enable high resolution reconstruction of cellular activity during a specified time window—at length scales (superresolution microscopy, electron microscopy) that are not accessible with currently available sensors. Although many of the tools we are developing have applications in neuroscience, these strategies and techniques can be applied to fundamental cellular physiology. Additionally, my research program places a heavy emphasis on synthetic chemistry and molecular design to achieve our goals. I anticipate we will uncover fundamental insights in areas related to photoinduced electron transfer, supramolecular chemistry, physical organic chemistry, and biophysics as we design and develop these new tools.

抽象的 用于可视化细胞生理学的新化学工具 细胞膜是所有细胞生命体共有的唯一细胞器，离子分布不均匀。 和化学物质穿过质膜导致产生电化学 电位以及膜电位或电压的快速变化驱动了独特的生理学 然而，所有细胞，甚至非兴奋细胞，都具有兴奋性。 膜电位，越来越多的证据支持膜电位在控制中的作用 基础细胞生理学，例如细胞周期、迁移、增殖和分化 尽管膜电位对两者的细胞生理学起着核心作用。 可兴奋和不可兴奋细胞，我们对这些系统中膜电位的理解仍然存在 不完整，很大程度上是由于缺乏研究具有高空间和高通量的细胞生理学的工具 膜电位的时间分辨率测量依赖于高侵入性、低通量的直接测量。 通过电极（膜片钳）或间接监测下游效应记录电压 我们建议利用合成有机物的力量来测量膜电位。 化学设计荧光电压传感器来探测神经元的膜电位动态 除了非兴奋性细胞之外，我们正在以补充方法开发心肌细胞。 基于小分子的活性积分器，可随时间积分 Ca2+ 瞬变，以实现高分辨率 在指定时间窗口内以长度尺度重建细胞活动（超分辨率 目前可用的传感器无法实现的显微镜、电子显微镜）。 尽管我们正在开发的许多工具都应用于神经科学，但这些策略和 此外，我的研究项目还可以应用于基础细胞生理学。 我预计我们会高度重视合成化学和分子设计来实现我们的目标。 揭示光诱导电子转移、超分子相关领域的基本见解 我们设计和开发这些新工具时涉及化学、物理有机化学和生物物理学。

项目成果

Mild and scalable synthesis of phosphonorhodamines.

• DOI: 10.1039/d3sc02590j
• 发表时间: 2023-10-25
• 期刊: CHEMICAL SCIENCE
• 影响因子: 8.4
• 作者: Turnbull, Joshua L.;Golden, Ryan P.;Benlian, Brittany R.;Henn, Katharine M.;Lipman, Soren M.;Miller, Evan W.
• 通讯作者: Miller, Evan W.

Simultaneous recording of multiple cellular signaling events by frequency- and spectrally-tuned multiplexing of fluorescent probes.

• DOI: 10.7554/elife.63129
• 发表时间: 2021-12-03
• 期刊: eLife
• 影响因子: 7.7
• 作者: Kierzek M;Deal PE;Miller EW;Mukherjee S;Wachten D;Baumann A;Kaupp UB;Strünker T;Brenker C
• 通讯作者: Brenker C

Pulsus Alternans in Cardiogenic Shock Recapitulated in Single Cell Fluorescence Imaging of a Patient's Cardiomyocyte.

• DOI: 10.1161/circheartfailure.121.008855
• 发表时间: 2022-03
• 期刊: Circulation. Heart failure
• 作者: Moreno JD;Bhagavan D;Li A;Gerstner NC;Miller EW;Huebsch N;Cresci S;Silva JR
• 通讯作者: Silva JR

Fluorescent indicators for imaging membrane potential of organelles.

• DOI: 10.1016/j.cbpa.2022.102203
• 发表时间: 2022-12
• 期刊: Current opinion in chemical biology
• 影响因子: 7.8

Heart Muscle Microphysiological System for Cardiac Liability Prediction of Repurposed COVID-19 Therapeutics.

• DOI: 10.3389/fphar.2021.684252
• 发表时间: 2021
• 期刊: Frontiers in pharmacology
• 影响因子: 5.6
• 作者: Charrez B;Charwat V;Siemons BA;Goswami I;Sakolish C;Luo YS;Finsberg H;Edwards AG;Miller EW;Rusyn I;Healy KE
• 通讯作者: Healy KE