Electrophysiologic Analysis of RIM Function in Presynaptic Plasticity

突触前可塑性 RIM 功能的电生理分析

• 批准号: 7693678
• 负责人: ROBERT C MALENKA
• 金额: $ 46.57万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7693678
• 关键词: Acute; Address; Animals; Behavior; Behavioral; Binding; Biochemical; Biological; Brain; Cells; Cerebellum; Chromosome Pairing; Class; Collaborations; Computer Simulation; Cyclic AMP-Dependent Protein Kinases; Disease; Fiber; Gene Deletion; Generations; Genes; Health; Hippocampal Mossy Fibers; Hippocampus (Brain); Individual; Knock-out; Knockout Mice; Knowledge; Lead; Length; Long-Term Potentiation; Mediating; Mediator of activation protein; Modification; Molecular; Mus; Mutant Strains Mice; Mutate; Neurons; Numbers; Phosphorylation; Physiological; Principal Investigator; Program Research Project Grants; Property; Protein Binding Domain; Protein Isoforms; Protein Kinase; Protein Overexpression; Proteins; Purkinje Cells; Range; Role; Serine; Signal Transduction; Site; Slice; Staging; Stimulus; Structure; Synapses; Synaptic Vesicles; Synaptic plasticity; Techniques; Testing; Training; Viral; Work; base; design; experience; expression vector; granule cell; hippocampal pyramidal neuron; in vivo; interest; member; mossy fiber; motor learning; mouse model; mutant; neural circuit; neuropsychiatry; neurotransmitter release; postsynaptic; presynaptic; prevent; programs; protein protein interaction; relating to nervous system; research study; response; size; stellate cell; synaptic function; tool

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.

项目#2：突触前可塑性中 RIM 功能的电生理分析 阐明突触可塑性的分子基础和生理意义将导致更多的研究 对依赖经验的神经回路修改的深入理解 健康和疾病的弹性。关于长突触后形式的机制已经了解很多 具有可塑性。然而，相比之下，人们对长期的潜在机制知之甚少。 突触前可塑性的持久形式。在这个提案中，我们重点关注理解突触功能 突触前活性区蛋白 RIM 的一类，因为它们需要以突出的形式参与 突触前 LTP 及其在基础神经递质释放和短期可塑性中的附加作用。 RIM 具有多个蛋白质结合域，可与突触小泡的关键成分和活性物质相互作用。 区。在目标 1 中，我们将通过以下方式评估 RIM 多种蛋白质相互作用的生理意义： 试图挽救缺乏 RIM 的自动培养海马神经元的突触异常 破坏单个蛋白质相互作用的突变 RIM。在目标 2 中，我们将评估以下人员的功能角色： 通过检查自动培养神经元中的突触功能，研究了几种不同的 RIM 亚型，其中这些亚型是 缺失或过度表达。在目标 3 中，我们将使用敲入小鼠模型来测试以下功能的意义： 通过评估假设突触前 LTP 所需的关键丝氨酸残基 (S413A) 发生突变 自动培养的神经元和急性海马和小脑切片中的突触功能 这些突变小鼠。在目标 4 中，我们将进一步描述突触前 LTP 的几个特征： 项目 4 中的小脑研究了其对运动学习的假设贡献。采取 总之，这些研究将有助于阐明多种形式的突触前可塑性和 能够生成有助于检查其功能意义的工具 行为层面。通过定义 RIM 介导突触可塑性和行为的分子相互作用，我们 将生成对于治疗广泛疾病所需的靶向这些蛋白质至关重要的信息 一系列神经精神疾病。