Electrophysiological and mechanical activity of cells triggered and traced on the level of single receptor-ligand interactions

在单一受体-配体相互作用水平上触发和追踪细胞的电生理和机械活动

• 批准号: 90621944
• 负责人: Dr. Martin Benoit
• 金额: --
• 依托单位: Lehrstuhl für Experimentelle Physik - Biophysik
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2008
• 资助国家: 德国
• 起止时间: 2007-12-31 至 2011-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/90621944?language=en
• 关键词: Electrophysiological; mechanical; activity; cells; triggered

The complexity of signaling pathways calls for several approaches and strategies to deduce the interconnections between the involved molecules and a measured reaction of the cell. In a majority of experiments the signaling receptors are stimulated by soluble ligands. But sometimes even a stimulus on a single receptor results in a strong cellular activity (e.g. a single photon can trigger a whole photoreceptor cell). Approaches that involve patch-clamp are precise in electrical stimulation and successful in tracing the dynamics of signaling processes since most of them include ion channel activity. Neuronal receptors often directly couple to ion channels. Approaches that focus on mechanical activity of cells still are less favored. Here we want to utilize the nano mechanical precision of an AFM to apply a stimulus to certain molecules on a cell and to record the cellular movement as a response to a stimulus combined with single channel recordings by simultaneous patch-clamp. This unique setup records electro-physiological and mechanical reactions of a cell to a given stimulus at high sampling rate and resolution. The interplay of mechanical and electrophysiological reactions of individual angiotensin II peptides interacting with its G-protein coupled receptor pathway and of individual glutamate molecules interacting with its ionotropic receptor are targeted in this approach for the first time.

信号通路的复杂性需要几种方法和策略来推断所涉及的分子和细胞的测量反应之间的相互联系。在大多数实验中，信号受体被可溶性配体刺激。但有时，即使对单个受体的刺激也会导致强烈的细胞活性（例如，单个光子可以触发整个感光细胞）。涉及膜片钳的方法在电刺激中是精确的，并且在跟踪信号传导过程的动力学方面是成功的，因为它们中的大多数包括离子通道活性。神经元受体通常直接偶联到离子通道。专注于细胞机械活动的方法仍然不太受欢迎。在这里，我们希望利用AFM的纳米机械精度来对细胞上的某些分子施加刺激，并记录细胞运动作为对刺激的响应，并通过同步膜片钳进行单通道记录。这种独特的设置以高采样率和分辨率记录细胞对给定刺激的电生理和机械反应。机械和电生理反应的相互作用，个别血管紧张素II肽与其G-蛋白偶联受体途径和个别谷氨酸分子与其离子型受体相互作用的相互作用，在这种方法中的第一次有针对性。