Optogenetic interrogation of neural circuits: technology for probing mammalian brain structures.
Optogenetic interrogation of neural circuits: technology for probing mammalian brain structures.
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DOI：
10.1038/nprot.2009.226
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发表时间：
2010-03
期刊：
影响因子：
14.8
通讯作者：
Deisseroth K
中科院分区：
文献类型：
--
作者：
Zhang F;Gradinaru V;Adamantidis AR;Durand R;Airan RD;de Lecea L;Deisseroth K
阐明复杂动物行为背后的神经基质依赖于精确的活动控制工具，以及兼容的读出方法。光遗传学的最新发展满足了这一需求，为系统神经科学开辟了新的可能性。甚至可以通过直接探测具有毫秒级、细胞类型特定的光学扰动的定义的电路元件的必要性和充分性来进行深层神经电路的询问，再加上适当的读出，例如电生理学、光学电路动态测量和哺乳动物的自由移动行为。在这里，我们详细收集了将微生物视蛋白基因递送到体内哺乳动物深层大脑结构的策略，以及将所得光学控制与兼容读数（电生理、光学和行为）集成的协议。沿着。这里描述的程序，从最初的病毒制备到系统级功能读出，可以在4-5周内完成。总之，这些方法可以帮助提供电路级的洞察力，在健康和疾病的复杂的哺乳动物行为的动力学。
Elucidation of the neural substrates underlying complex animal behaviors depends on precise activity control tools, as well as compatible readout methods. Recent developments in optogenetics have addressed this need, opening up new possibilities for systems neuroscience. Interrogation of even deep neural circuits can be conducted by directly probing the necessity and sufficiency of defined circuit elements with millisecond-scale, cell type-specific optical perturbations, coupled with suitable readouts such as electrophysiology, optical circuit dynamics measures and freely moving behavior in mammals. Here we collect in detail our strategies for delivering microbial opsin genes to deep mammalian brain structures in vivo, along with protocols for integrating the resulting optical control with compatible readouts (electrophysiological, optical and behavioral). The procedures described here, from initial virus preparation to systems-level functional readout, can be completed within 4–5 weeks. Together, these methods may help in providing circuit-level insight into the dynamics underlying complex mammalian behaviors in health and disease.