Enhanced Archaerhodopsin Fluorescent Protein Voltage Indicators.
Enhanced Archaerhodopsin Fluorescent Protein Voltage Indicators.
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DOI：
10.1371/journal.pone.0066959
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Schnitzer MJ
中科院分区：
文献类型：
--
作者：
Gong Y;Li JZ;Schnitzer MJ
神经科学的一个长期目标是开发一种技术，用于成像遗传定义的神经元子集的电压动态。跨膜电压光学传感器将加强从体外培养的单个神经元到清醒行为动物的神经元群体的神经活动研究。最近的进展已经确定了基于Arch的传感器是一种有前途的，遗传编码的荧光电压指示器，可以报告单一动作电位。野生型Arch对跨膜电压表现出亚毫秒级的荧光响应，但其光激活质子泵也对成像照明有响应。Arch突变体（Arch- d95n）没有光电流，但对电压瞬变的响应较慢，约为40 ms。在这里，我们提出了拱衍生电压传感器与交通信号，加强其定位到神经膜。我们还描述了Arch突变型传感器（Arch- een和-EEQ），它们比Arch- d95n具有更快的动力学和更大的荧光动态范围，并且在通常用于成像的照明强度下没有光电流。我们使用考虑到实验测量的光子散粒噪声和单动作电位光波形的信号检测理论框架，对这些电压传感器的尖峰检测保真度进行了基准测试。结果表明，与Arch-D95N相比，通过结合序列突变和增强的传输序列，新传感器的峰值检测保真度提高了近3倍。
A longstanding goal in neuroscience has been to develop techniques for imaging the voltage dynamics of genetically defined subsets of neurons. Optical sensors of transmembrane voltage would enhance studies of neural activity in contexts ranging from individual neurons cultured in vitro to neuronal populations in awake-behaving animals. Recent progress has identified Archaerhodopsin (Arch) based sensors as a promising, genetically encoded class of fluorescent voltage indicators that can report single action potentials. Wild-type Arch exhibits sub-millisecond fluorescence responses to trans-membrane voltage, but its light-activated proton pump also responds to the imaging illumination. An Arch mutant (Arch-D95N) exhibits no photocurrent, but has a slower, ~40 ms response to voltage transients. Here we present Arch-derived voltage sensors with trafficking signals that enhance their localization to the neural membrane. We also describe Arch mutant sensors (Arch-EEN and -EEQ) that exhibit faster kinetics and greater fluorescence dynamic range than Arch-D95N, and no photocurrent at the illumination intensities normally used for imaging. We benchmarked these voltage sensors regarding their spike detection fidelity by using a signal detection theoretic framework that takes into account the experimentally measured photon shot noise and optical waveforms for single action potentials. This analysis revealed that by combining the sequence mutations and enhanced trafficking sequences, the new sensors improved the fidelity of spike detection by nearly three-fold in comparison to Arch-D95N.
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DOI：
10.1073/pnas.86.2.529
发表时间：
1989-01-01
影响因子：
11.1
作者：
MARINETTI, T;SUBRAMANIAM, S;KHORANA, HG
通讯作者：
KHORANA, HG
影响因子：
16.2
作者：
Jin L;Han Z;Platisa J;Wooltorton JR;Cohen LB;Pieribone VA
通讯作者：
Pieribone VA
影响因子：
3.4
作者：
Mishina, Yukiko;Mutoh, Hiroki;Knoepfel, Thomas
通讯作者：
Knoepfel, Thomas
影响因子：
25
作者：
Boyden, ES;Zhang, F;Deisseroth, K
通讯作者：
Deisseroth, K
影响因子：
48
作者：
Kralj, Joel M.;Douglass, Adam D.;Hochbaum, Daniel R.;Maclaurin, Dougal;Cohen, Adam E.
通讯作者：
Cohen, Adam E.