Targets of low dose alcohol during cerebellar-driven behavior in mice

项目总结 小脑神经元是神经系统中对酒精最敏感的目标之一。在浓缩物 酒精会导致小脑功能障碍，仅略高于美国的法定驾车限制(17 Mm BAC)。 导致步态、平衡和运动协调能力受损，导致数千人受伤 以及每年的死亡人数。小脑切片的电生理学实验也有证据表明 较低浓度(10 MM)的酒精会改变小脑中多个神经元的功能。然而，我们的 对低剂量酒精在小脑中的作用的了解仍处于初级阶段。一个严重的障碍 该领域的进展是理解体外观察到的低剂量酒精的影响如何转化为 未麻醉的动物，并导致小脑驱动行为中的功能受损。这样做的目的是 建议是通过第一个测量低剂量酒精对人体的影响来克服这一先前的限制。 行为正常的小鼠的小脑。小鼠将接受小脑依赖的眨眼条件反射训练，因为 我们的初步数据表明，低剂量的酒精(&lt；5 mMBAC)会损害这项任务的表现。我们的 一般的策略是在眨眼条件反射过程中寻找低剂量酒精的细胞特异性靶点，通过 记录和操纵三个不同处理阶段的小脑回路活动：输出 中间阶段，由间位核神经元处理(特定目标1)，中间阶段，由 浦肯野细胞(特异性目标2)和输入阶段，由颗粒细胞加工(特异性目标3)。建议数 实验将开辟新的天地，大大推进这一领域：通过结合眨眼的简单性 利用光遗传学、电生理学和双光子钙成像的新技术进行调节 该项目将首次展示低剂量酒精、小脑和行为之间的机械联系。

PROJECT SUMMARY Cerebellar neurons are among the most sensitive targets of alcohol in the nervous system. At concentrations just above the legal limit for driving in the United States (>17mM BAC), alcohol causes cerebellar dysfunction, leading to impairments in gait, balance and motor coordination that are responsible for thousands of injuries and deaths every year. There is also evidence from electrophysiology experiments in cerebellar slices that at lower concentrations (<10mM), alcohol alters the function of multiple neurons in the cerebellum. However, our understanding of the action of low doses of alcohol in the cerebellum is still in its infancy. A critical barrier to progress in the field has been to understand how the effects of low dose alcohol observed in vitro translate to the unanesthetized animal and lead to impaired function during cerebellar-driven behaviors. The goal of this proposal is to overcome this prior limitation, by being the first to measure the impact of low doses of alcohol in the cerebellum of behaving mice. Mice will be trained in cerebellum-dependent eyeblink conditioning because our preliminary data indicates that low doses of alcohol (<5mM BAC) impair performance in this task. Our general strategy is to search for cell-specific targets of low dose alcohol during eyeblink conditioning, by recording and manipulating the activity of cerebellar circuits at three different stages of processing: the output stage, processed by neurons of the interpositus nucleus (specific aim 1), the intermediate stage, processed by Purkinje cells (specific aim 2), and the input stage, processed by granule cells (specific aim 3). The proposed experiments will break new ground and advance the field substantially: By combining the simplicity of eyeblink conditioning with new technologies for optogenetics, electrophysiology, and two-photon calcium imaging, this project will provide a first look at the mechanistic links between low dose alcohol, the cerebellum, and behavior.