通过膜片钳实验对微生物视紫红质进行电生理特性分析。

Electrophysiological Characterization of Microbial Rhodopsins by Patch-Clamp Experiments.

机构信息

Advanced Optogenes Group, Institute for Auditory Neuroscience and InnerEarLab, University Medical Center Gottinggen, Gottingen, Germany.

Cluster of Excellence "Multiscale Bioimaging: From Molecular Machines to Networks of Excitable Cells" (MBExC), University of Goettingen, Goettingen, Germany.

出版信息

Methods Mol Biol. 2022;2501:277-288. doi: 10.1007/978-1-0716-2329-9_13.

DOI:10.1007/978-1-0716-2329-9_13

PMID:35857233

Abstract

Optogenetics is of key importance for progress in basic neuroscience research and the development of innovative future medical treatments. In particular, the use of microbial rhodopsins enables remote control of excitable-cell activity by light. The electrophysiological characterization of microbial rhodopsins is inevitable for the development of variants, which further advance optogenetic applications. Therefore, we provide a detailed description of the application of the patch-clamp method for the electrophysiological characterization of microbial rhodopsins. Here we describe the investigation of light sensitivity, wavelength- and voltage-dependence, photocurrent inactivation, kinetics, and ion selectivity.

摘要

光遗传学对于基础神经科学研究的进展和创新未来医学治疗方法的发展至关重要。特别是，微生物视蛋白的使用使得通过光来远程控制可兴奋细胞的活性成为可能。微生物视蛋白变体的开发离不开对其进行电生理学特性的表征，这进一步推动了光遗传学的应用。因此，我们提供了详细描述应用膜片钳技术对微生物视蛋白进行电生理学特性表征的方法。在这里，我们描述了对光敏感性、波长和电压依赖性、光电流失活、动力学和离子选择性的研究。