A calcium- and light-gated switch to induce gene expression in activated neurons
A calcium- and light-gated switch to induce gene expression in activated neurons
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DOI：
10.1038/nbt.3902
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发表时间：
2017-09-01
影响因子：
46.9
通讯作者：
Kwon, Hyung-Bae
中科院分区：
文献类型：
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作者：
Lee, Dongmin;Hyun, Jung Ho;Kwon, Hyung-Bae
尽管光遗传学最近取得了进展，但在特定神经元群体中操纵基因表达仍然具有挑战性。我们提出了一个双蛋白开关系统，钙光，翻译神经元活性介导的钙信号转化为基因表达的光依赖性的方式。在培养的神经元和脑切片中，我们表明，钙光驱动的报告EGFP的表达具有高时空分辨率，只有在蓝光和钙的存在下。通过病毒载体将Cal-Light组分递送到小鼠的运动皮层，标记与学习性压迫行为相关的兴奋性和抑制性神经元的子集。通过使用Cal-Light来驱动对黄光有反应的抑制性受体盐视紫红质（eNpHR）的表达，我们暂时抑制了神经元的压迫行为，证实了标记的神经元介导了该行为。因此，Cal-Light能够以高时空精度解剖复杂哺乳动物行为背后的神经回路。
Despite recent advances in optogenetics, it remains challenging to manipulate gene expression in specific populations of neurons. We present a dual-protein switch system, Cal-Light, that translates neuronal-activity-mediated calcium signaling into gene expression in a light-dependent manner. In cultured neurons and brain slices, we show that Cal-Light drives expression of the reporter EGFP with high spatiotemporal resolution only in the presence of both blue light and calcium. Delivery of the Cal-Light components to the motor cortex of mice by viral vectors labels a subset of excitatory and inhibitory neurons related to learned lever-pressing behavior. By using Cal-Light to drive expression of the inhibitory receptor halorhodopsin (eNpHR), which responds to yellow light, we temporarily inhibit the lever-pressing behavior, confirming that the labeled neurons mediate the behavior. Thus, Cal-Light enables dissection of neural circuits underlying complex mammalian behaviors with high spatiotemporal precision.