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

项目摘要 小脑神经元是神经系统中对酒精最敏感的目标之一。浓度 仅高于美国驾驶的法律的限制（> 17mMBAC），酒精引起小脑功能障碍， 导致步态、平衡和运动协调障碍，导致数千起伤害 和死亡人数。小脑切片的电生理学实验也有证据表明， 低浓度（<10 mM）的酒精会改变小脑中多个神经元的功能。但我们的 对小剂量酒精对小脑作用的了解仍处于初期阶段。一个关键的障碍， 该领域的进展是理解在体外观察到的低剂量酒精的作用如何转化为 在未麻醉的动物中，导致小脑驱动行为的功能受损。这个目标 该提案旨在克服这一先前的限制，通过第一个测量低剂量酒精对 行为正常的老鼠的小脑。小鼠将接受小脑依赖性眨眼条件反射训练， 我们的初步数据表明，低剂量的酒精（<5mM BAC）会损害这项任务的表现。我们 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， 记录和操纵小脑回路在三个不同处理阶段的活动：输出 阶段，由间位核的神经元处理（特定目的1），中间阶段，由 浦肯野细胞（特异性目的2）和输入阶段，由颗粒细胞处理（特异性目的3）。拟议 实验将开辟新天地，大大推进该领域：通过结合眨眼的简单性， 光遗传学、电生理学和双光子钙成像的新技术， 该项目将提供低剂量酒精，小脑和行为之间的机械联系的第一眼。

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.