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 可视化细胞生理的新化学工具 细胞膜是唯一通过细胞寿命的变体共享的细胞器。离子的不平等分布 质膜上的化学物种会产生电化学 该膜电位或电压的潜力和快速变化推动了独特的生理学 令人兴奋的细胞等神经元和心肌细胞。但是，所有细胞，甚至是不可观的细胞都具有 膜电位和安装证据支持膜潜能控制的作用 基本的细胞生理学 - 例如，细胞周期，迁移，增殖和分化 - 不可驱行的细胞。尽管膜潜力在两者的细胞生理学上具有核心作用 令人兴奋且非驱动的细胞，我们对这些系统中的膜潜力的理解仍然存在 不完整，在很大程度上是由于缺乏用于研究细胞生理学的工具，并具有很高的空间和 临时分辨率。膜电位的测量取决于高度侵入性，低通量直接 通过电极记录电压（贴片夹）或间接监视下游效应 通过成像（CA2+成像）的膜电位。我们建议利用合成有机物的力量 化学以设计荧光电压传感器，以探测神经元和 心肌细胞，除了不可驱行的细胞。在一种完整的方法中，我们正在发展 基于小分子的活性积分器，这些积分器会随着时间的推移整合Ca2+瞬变以实现高分辨率 在指定的时间窗口中重建细胞活动的时间（长度分辨率） 显微镜，电子显微镜），当前可用的传感器无法访问。 尽管我们正在开发的许多工具在神经科学中都有应用，但这些策略和 技术可以应用于基本的细胞生理学。此外，我的研究计划位置 强调合成化学和分子设计以实现我们的目标。我期望我们会 发现与光诱导的电子转移，超分子有关的领域的基本见解 当我们设计和开发这些新工具时，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