一种能够分辨动作电位的荧光、基因编码电压探针。

A fluorescent, genetically-encoded voltage probe capable of resolving action potentials.

机构信息

Department of Cell Biology and Neuroscience, Montana State University, Bozeman, Montana, United States of America.

出版信息

PLoS One. 2012;7(9):e43454. doi: 10.1371/journal.pone.0043454. Epub 2012 Sep 6.

Abstract

There is a pressing need in neuroscience for genetically-encoded, fluorescent voltage probes that can be targeted to specific neurons and circuits to allow study of neural activity using fluorescent imaging. We created 90 constructs in which the voltage sensing portion (S1-S4) of Ciona intestinalis voltage sensitive phosphatase (CiVSP) was fused to circularly permuted eGFP. This led to ElectricPk, a probe that is an order of magnitude faster (taus 1-2 ms) than any currently published fluorescent protein-based voltage probe. ElectricPk can follow the rise and fall of neuronal action potentials with a modest decrease in fluorescence intensity (0.7% ΔF/F). The probe has a nearly linear fluorescence/membrane potential response to both hyperpolarizing and depolarizing steps. This is the first probe based on CiVSP that captures the rapid movements of the voltage sensor, suggesting that voltage probes designed with circularly permuted fluorescent proteins may have some advantages.

摘要

神经科学领域迫切需要基因编码的荧光电压探针,这些探针可以靶向特定的神经元和回路,从而使用荧光成像来研究神经活动。我们构建了 90 种构建体,其中将文昌鱼电压敏感磷酸酶(CiVSP)的电压感应部分(S1-S4)与环化排列的 eGFP 融合。这导致了 ElectricPk,这是一种比任何当前发表的基于荧光蛋白的电压探针快一个数量级(tau1-2ms)的探针。ElectricPk 可以跟随神经元动作电位的上升和下降,荧光强度略有下降(0.7%ΔF/F)。该探针对超极化和去极化步骤的荧光/膜电位响应几乎呈线性。这是第一个基于 CiVSP 的可以捕捉电压传感器快速运动的探针,这表明使用环化排列的荧光蛋白设计的电压探针可能具有一些优势。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d222/3435330/f43464bac9bf/pone.0043454.g001.jpg

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