A fluorescent, genetically-encoded voltage probe capable of resolving action potentials.
A fluorescent, genetically-encoded voltage probe capable of resolving action potentials.
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DOI：
10.1371/journal.pone.0043454
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Hughes T
中科院分区：
文献类型：
--
作者：
Barnett L;Platisa J;Popovic M;Pieribone VA;Hughes T
神经科学迫切需要基因编码的荧光电压探针，可以靶向特定的神经元和电路，以允许使用荧光成像研究神经活动。我们创建了90个构建体，其中玻璃海鞘电压敏感磷酸酶（CiVSP）的电压感应部分（S1-S4）与环状排列的eGFP融合。这导致了ElectricPk，这是一种比任何目前公布的基于荧光蛋白的电压探针快一个数量级（τ 1-2 ms）的探针。ElectricPk可以跟随神经元动作电位的上升和下降，荧光强度适度降低（ΔF/F <0.7%）。该探针具有近线性的荧光/膜电位响应超极化和去极化步骤。这是第一个基于CiVSP的探针，它捕获了电压传感器的快速移动，这表明用循环排列的荧光蛋白设计的电压探针可能具有一些优势。
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.
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