Kodani Shota, Soya Shingo, Sakurai Takeshi
Department of Molecular Neuroscience and Integrative Physiology, Faculty of Medicine, Kanazawa University.
International Institute for Integrative Sleep Medicine (WPI-IIIS), University of Tsukuba.
J Vis Exp. 2019 Jun 19(148). doi: 10.3791/58613.
In recent years, optogenetics has been widely used in many fields of neuroscientific research. In many cases, an opsin, such as channel rhodopsin 2 (ChR2), is expressed by a virus vector in a particular type of neuronal cells in various Cre-driver mice. Activation of these opsins is triggered by application of light pulses which are delivered by laser or LED through optic cables, and the effect of activation is observed with very high time resolution. Experimenters are able to acutely stimulate neurons while monitoring behavior or another physiological outcome in mice. Optogenetics can enable useful strategies to evaluate function of neuronal circuits in the regulation of sleep/wakefulness states in mice. Here we describe a technique for examining the effect of optogenetic manipulation of neurons with a specific chemical identity during electroencephalogram (EEG) and electromyogram (EMG) monitoring to evaluate the sleep stage of mice. As an example, we describe manipulation of GABAergic neurons in the bed nucleus of the stria terminalis (BNST). Acute optogenetic excitation of these neurons triggers a rapid transition to wakefulness when applied during NREM sleep. Optogenetic manipulation along with EEG/EMG recording can be applied to decipher the neuronal circuits that regulate sleep/wakefulness states.
近年来，光遗传学已广泛应用于神经科学研究的许多领域。在许多情况下，一种视蛋白，如通道视紫红质2（ChR2），由病毒载体在各种Cre驱动小鼠的特定类型神经元细胞中表达。这些视蛋白的激活是通过激光或发光二极管通过光缆传递的光脉冲来触发的，并且以非常高的时间分辨率观察激活的效果。实验者能够在监测小鼠行为或其他生理结果的同时急性刺激神经元。光遗传学能够提供有用的策略来评估小鼠睡眠/觉醒状态调节中神经回路的功能。在此，我们描述一种在脑电图（EEG）和肌电图（EMG）监测期间检查具有特定化学特性的神经元的光遗传学操纵对小鼠睡眠阶段影响的技术。作为一个例子，我们描述对终纹床核（BNST）中GABA能神经元的操纵。在非快速眼动睡眠期间应用时，这些神经元的急性光遗传学兴奋会触发快速转变为觉醒状态。光遗传学操纵与EEG/EMG记录一起可用于破译调节睡眠/觉醒状态的神经回路。
