High-performance genetically targetable optical neural silencing by light-driven proton pumps.
High-performance genetically targetable optical neural silencing by light-driven proton pumps.
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以时间上精确的方式沉默遗传上指定的神经元的活动的能力将开启研究特定细胞类别在神经计算、行为和病理中的因果作用的能力。在这里，我们表明光驱动外向质子泵类成员可以介导非常强大、安全、多色的神经活动沉默。来自Halorubrum sodomense的基因archaerhodopsin-3（Arch）当在小鼠皮层中病毒表达并用黄光照射时，能够使清醒大脑中的神经元接近100%沉默。Arch在低光功率下介导数百皮安的电流，并在体内容易实现的光功率下支持接近900 pA的神经沉默电流。此外，Arch自发地从光依赖性失活中恢复，不像光驱动的氯离子泵在响应光时进入持久的失活状态。Arch的这些特性适合于在行为相关的时间尺度上介导显著脑容量的光学沉默。神经元中的Arch功能耐受良好，因为Arch照明产生的pH偏移通过自限机制最小化至与通道视紫红质或自然尖峰放电介导的水平相当的水平。为了强调质子泵生态和基因组多样性如何支持新的创新，我们证明了来自真菌Leptosphaeria maculans（Mac）的蓝绿光驱动质子泵在神经元中表达时，可以通过蓝光实现神经沉默，从而与其他开发的试剂一起实现蓝光与红光对两个神经群体的独立沉默。因此，光驱动质子泵代表了一种高性能和极其通用的“光遗传学”电压和离子调制器，它将广泛地支持新的神经科学，生物学，神经学和精神病学研究。
The ability to silence the activity of genetically specified neurons in a temporally precise fashion would open up the ability to investigate the causal role of specific cell classes in neural computations, behaviors, and pathologies. Here we show that members of the class of light-driven outward proton pumps can mediate very powerful, safe, multiple-color silencing of neural activity. The gene archaerhodopsin-3 (Arch) from Halorubrum sodomense enables near-100% silencing of neurons in the awake brain when virally expressed in mouse cortex and illuminated with yellow light. Arch mediates currents of several hundred picoamps at low light powers, and supports neural silencing currents approaching 900 pA at light powers easily achievable in vivo. In addition, Arch spontaneously recovers from light-dependent inactivation, unlike light-driven chloride pumps that enter long-lasting inactive states in response to light. These properties of Arch are appropriate to mediate the optical silencing of significant brain volumes over behaviourally-relevant timescales. Arch function in neurons is well tolerated because pH excursions created by Arch illumination are minimized by self-limiting mechanisms to levels comparable to those mediated by channelrhodopsins or natural spike firing. To highlight how proton pump ecological and genomic diversity may support new innovation, we show that the blue-green light-drivable proton pump from the fungus Leptosphaeria maculans (Mac) can, when expressed in neurons, enable neural silencing by blue light, thus enabling alongside other developed reagents the potential for independent silencing of two neural populations by blue vs. red light. Light-driven proton pumps thus represent a high-performance and extremely versatile class of “optogenetic” voltage and ion modulator, which will broadly empower new neuroscientific, biological, neurological, and psychiatric investigations.
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