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: 8132217
• 负责人: ROBERT C MALENKA
• 金额: $ 40.51万
• 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/8132217
• 关键词: Acute; Address; Animals; Behavior; Behavioral; Binding; Biochemical; Biological; Brain; Cells; Cerebellum; 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; Phosphorylation; Physiological; Principal Investigator; Program Research Project Grants; Property; Protein Binding Domain; Protein Isoforms; Protein Kinase; Proteins; Purkinje Cells; 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; overexpression; postsynaptic; presynaptic; prevent; programs; protein protein interaction; relating to nervous system; research study; response; 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 对神经回路修改的复杂理解，这是经验依赖的基础 健康和疾病中的弹性。关于突触后长突触形式的机制， 铸造塑性然而，相比之下，对长期的潜在机制知之甚少。 突触前可塑性的持久形式。在这个建议中，我们专注于了解突触的功能， 一类突触前活动区蛋白，RIMs，因为它们需要以突出的形式参与 突触前LTP及其在基底神经递质释放和短时程可塑性中的作用。 RIM具有几个蛋白结合结构域，其与突触囊泡的关键组分相互作用， 区界限在目标1中，我们将通过以下方式评估RIM的不同蛋白质相互作用的生理意义： 试图挽救缺乏RIM的自发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 几种不同的RIM亚型通过检查突触功能的autaptic培养的神经元，其中这些是 缺失或过度表达。在目标3中，使用敲入小鼠模型，我们将测试 突变一个关键的丝氨酸残基（S413A），假设是突触前LTP所需的，通过评估 在自发性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， 在项目4的背景下，小脑检查其对运动学习的假定贡献。采取 总之，这些研究将有助于阐明突触前可塑性的多种形式的分子基础， 能够生成工具，以便于在 行为层面。通过定义介导突触可塑性和行为的RIM分子相互作用， 将产生的信息，将是关键的目标，这些蛋白质的治疗所需的广泛的 一系列神经精神疾病。