Rhodopsin optogenetic toolbox v2.0 for light-sensitive excitation and inhibition in Caenorhabditis elegans.
Rhodopsin optogenetic toolbox v2.0 for light-sensitive excitation and inhibition in Caenorhabditis elegans.
复制标题
DOI：
10.1371/journal.pone.0191802
复制
发表时间：
2018
期刊：
影响因子：
3.7
通讯作者：
Liewald JF
中科院分区：
文献类型：
--
作者：
Bergs A;Schultheis C;Fischer E;Tsunoda SP;Erbguth K;Husson SJ;Govorunova E;Spudich JL;Nagel G;Gottschalk A;Liewald JF
在光遗传学中，视紫红质被确立为操纵神经元活动的光驱动工具。然而，在使用通道视紫红质（ChR）的长期光刺激期间，脱敏可以降低效果。此外，在缺乏足够的内源性浓度的模型系统中，对发色团全反式视黄醛（ATR）连续存在的要求限制了其适用性。我们测试了已知的，工程和特点的新变种的去极化和超极化视紫红质秀丽隐杆线虫。ChR 2变体组合了先前描述的点突变，其可以协同作用以实现延长的刺激。在短暂的光脉冲之后，ChR 2（C128 S; H134 R）诱导肌肉激活数分钟或甚至数小时（“Quint”：ChR 2（C128 S; L132 C; H134 R; D156 A; T159 C）），因此具有比先前描述的变体更长的开放状态寿命。此外，与阶跃函数视蛋白ChR 2（C128 S）相比，ATR去除后的稳定性增加。双突变体C128 S; H134 R和H134 R; D156 C能够在重复刺激期间增加效应。我们还测试了新的超偏振器（ACR 1、ACR 2、ACR 1（C102 A）、ZipACR）。特别是ACR 1和ACR 2在行为测定中显示出强烈的作用，并且具有快速动力学的非常大的电流。总之，我们引入了高度光敏的光遗传学工具，绕过了以前的缺点，从而构成了具有高效率和快速动力学的新工具，允许更好的重复刺激或研究C中延长的神经元活动状态。elegans线虫and，possibly可能，other systems系统.
In optogenetics, rhodopsins were established as light-driven tools to manipulate neuronal activity. However, during long-term photostimulation using channelrhodopsin (ChR), desensitization can reduce effects. Furthermore, requirement for continuous presence of the chromophore all-trans retinal (ATR) in model systems lacking sufficient endogenous concentrations limits its applicability. We tested known, and engineered and characterized new variants of de- and hyperpolarizing rhodopsins in Caenorhabditis elegans. ChR2 variants combined previously described point mutations that may synergize to enable prolonged stimulation. Following brief light pulses ChR2(C128S;H134R) induced muscle activation for minutes or even for hours (‘Quint’: ChR2(C128S;L132C;H134R;D156A;T159C)), thus featuring longer open state lifetime than previously described variants. Furthermore, stability after ATR removal was increased compared to the step-function opsin ChR2(C128S). The double mutants C128S;H134R and H134R;D156C enabled increased effects during repetitive stimulation. We also tested new hyperpolarizers (ACR1, ACR2, ACR1(C102A), ZipACR). Particularly ACR1 and ACR2 showed strong effects in behavioral assays and very large currents with fast kinetics. In sum, we introduce highly light-sensitive optogenetic tools, bypassing previous shortcomings, and thus constituting new tools that feature high effectiveness and fast kinetics, allowing better repetitive stimulation or investigating prolonged neuronal activity states in C. elegans and, possibly, other systems.
登录
查看更多内容
影响因子：
11.4
作者：
FIRE, A
通讯作者：
FIRE, A
影响因子：
25
作者：
Boyden, ES;Zhang, F;Deisseroth, K
通讯作者：
Deisseroth, K
影响因子：
3.5
作者：
Bruun, Sara;Naumann, Hendrik;Hildebrandt, Peter
通讯作者：
Hildebrandt, Peter
影响因子：
3.7
作者：
Husson, Steven J.;Liewald, Jana F.;Gottschalk, Alexander
通讯作者：
Gottschalk, Alexander
影响因子：
4.6
作者：
Govorunova EG;Sineshchekov OA;Rodarte EM;Janz R;Morelle O;Melkonian M;Wong GK;Spudich JL
通讯作者：
Spudich JL