Electrophysiologic Analysis of RIM Function in Presynaptic Plasticity

项目2：RIM在突触前可塑性中作用的电生理学分析 阐明突触可塑性的分子基础和生理意义将导致更多的 对作为经验依赖基础的神经回路改变有深入的了解 在健康和疾病中都有弹性。关于Long突触后形式的机制已知很多 铸态可塑性。然而，相比之下，人们对Long的潜在机制知之甚少 持久的突触前可塑性。在这项建议中，我们重点了解一个突触功能 一类突触前活动区蛋白，RIMS，因为它们需要以显著的形式参与 突触前LTP及其在基础神经递质释放和短期可塑性中的额外作用。 RIMS有几个蛋白结合域，与突触小泡的关键成分相互作用，并具有活性 区域。在目标1中，我们将通过以下方式评估RIM的各种蛋白质相互作用的生理学意义 自体培养缺环海马神经元突触异常的修复 破坏个体蛋白质相互作用的突变轮缘。在目标2中，我们将评估 通过检测自体培养神经元的突触功能，发现了几种不同的RIM亚型 缺席的或过度表达的。在目标3中，使用敲门小鼠模型，我们将测试 通过评估突变突触前LTP所需的关键丝氨酸残基(S413A) 自体培养神经元和急性脑片的突触功能 这些突变的小鼠。在目标4中，我们将进一步描述突触前LTP的几个特征 小脑在项目4的背景下，考察其对运动学习的假设贡献。已被占用 总之，这些研究将有助于阐明多种形式的突触前可塑性和 支持工具的生成，以便于在 行为层面。通过定义RIMS的分子相互作用来调节突触的可塑性和行为，我们 将产生对靶向这些蛋白质至关重要的信息，这些蛋白质是治疗广泛的 一系列神经精神疾病。

Project #2: Electrophysiologic Analysis of RIM Function in Presynaptic Plasticity Elucidating the molecular basis and physiological significance of synaptic plasticity will lead to a more sophisticated understanding of the neural circuit modifications which underlie experience-dependent olasticity in both health and disease. Much is known about the mechanisms of postsynaptic forms of long asting plasticity. By comparison, however, relatively little is known about the underlying mechanisms of long lasting forms of presynaptic plasticity. In this proposal we focus on understanding the synaptic functions of a class of presynaptic, active zone proteins, RIMs, because of their required involvement in a prominent form of presynaptic LTP and their additional roles in basal neuretransmitter release and short-term plasticity. RIMs have several protein binding domains that interact with key components of synaptic vesicles and active zones. In Aim 1, we will evaluate the physiologic significance of RIM's diverse protein interactions by attempting to rescue the synaptic abnormalities of autaptic cultured hippocampal neurons lacking RIMs with mutant RIMs that disrupt individual protein interactions. In Aim 2, we will evaluate the functional roles of several different RIM isoforms by examining synaptic function in autaptic cultured neurons in which these are absent or overexpressed. In Aim 3, using a knockin mouse model, we will test the functional significance of mutating a key serine residue (S413A) hypothesized to be required for presynaptic LTP by evaluating synaptic function in autaptic cultured neurons and acute hippocampal and cerebellar slices prepared from these mutant mice. In Aim 4, we will further characterize several features of presynaptic LTP in the cerebellum in the context of Project 4 which examines its postulated contribution to motor learning. Taken together, these studies will help elucidate the molecular basis of multiple forms of presynaptic plasticity and enable the generation of tools that will facilitate the examination of their functional significance at the behavioral level. By defining RIMs' molecular interactions that mediate synaptic plasticity and behavior, we will generate information that will be critical for targeting these proteins as needed for the treatment of a wide range of neuropsychiatric diseases.