Synaptic Analysis of Neuroligin1 function

Neuroligin1 功能的突触分析

• 批准号: 7676907
• 负责人: Marc V Fuccillo
• 金额: $ 5.01万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7676907
• 关键词: Acute; Address; Binding; Biochemical; Biological Assay; Biological Models; Biological Process; Brain; Cells; Disease; Excitatory Postsynaptic Potentials; Excitatory Synapse; Future; Genes; Genetic; Goals; Hippocampus (Brain); In Vitro; Individual; Injection of therapeutic agent; Intervention; Kinetics; Knockout Mice; Knowledge; Link; Long-Term Depression; Long-Term Potentiation; Mediating; Mediator of activation protein; Mental disorders; Molecular; Molecular Biology; Mus; Mutation; Neurons; Output; Physiologic pulse; Physiology; Point Mutation; Preparation; Probability; Proteins; Protocols documentation; Pyramidal Cells; Role; Slice; Synapses; Synaptic Transmission; Synaptic plasticity; Testing; Whole-Cell Recordings; Work; autism spectrum disorder; developmental disease; extracellular; ifenprodil; in vivo; loss of function; mutant; neuropsychiatry; postnatal; postsynaptic; research study; synaptic function; transmission process; voltage

DESCRIPTION (provided by candidate): The overarching goal of this proposal is to explore the mature synaptic function of Neuroliginl, a synapse- specific protein implicated in several autism spectrum disorders (ASDs). Molecular biology and mouse genetics will be used together with synaptic physiology to better understand the normal biological function of NL1 and the possible contributions of mutations in this gene to abnormal synaptic function in neuropsychiatric disorders. In addition to providing potential model systems for the study of ASDs, results from this work will contribute significantly to the limited understanding of NL function at mature synapses. In doing so, they may provide new targets for future molecular interventions in psychiatric disorders. Previous work from the hippocampus of NL1 knockout mice demonstrated a 50% reduction in the NMDAR/AMPAR ratio at Schaffer collateral/CA1 synapses. My first two specific aims will specifically address this observation by separately characterizing alterations in AMPAR- and NMDAR-mediated currents in control and NL1 mutant acute hippocampal slice preparations. I will follow these experiments by testing whether NL1 functions in NMDAR-dependent long-term potentiation (LTP) or long-term depression (LTD) at Schaffer collateral-CA1 synapses. Finally, to assess whether NL1 control of synaptic transmission has a pre- or post- synaptic locus, I will use postnatal lentiviral injection in vivo to "rescue" individual cells in NL1 KO mice with point mutants that disrupt either extracellular (3-neurexin binding or intracellular binding to PSD-95. The results of these experiments should both compliment our current in vitro knowledge while enhancing our understanding of the function of NL1 in the mature hippocampal circuit. Autism spectrum disorders (ASDs) comprise a heterogeneous group of neuro-developmental disorders that are highly heritable. Neuroliginl, a molecule found at synapses, has been implicated in familial ASDs. This proposal seeks to better understand the function of NL1 in the mature brain so that disorders resulting from abnormalities in this gene can one day be ameliorated.

描述（由候选人提供）：该提案的总体目标是探索神经胶质素1的成熟突触功能，神经胶质素1是一种涉及几种自闭症谱系障碍（ASD）的突触特异性蛋白。分子生物学和小鼠遗传学将与突触生理学一起使用，以更好地了解NL 1的正常生物学功能以及该基因突变对神经精神疾病中异常突触功能的可能贡献。除了为ASD的研究提供潜在的模型系统外，这项工作的结果将大大有助于对成熟突触NL功能的有限理解。通过这样做，他们可能会为未来精神疾病的分子干预提供新的目标。NL 1基因敲除小鼠海马的先前研究表明，Schaffer侧支/CA 1突触的NMDAR/AMPAR比率降低了50%。我的前两个具体目标将专门解决这一观察分别表征AMPAR和NMDAR介导的电流在控制和NL 1突变急性海马切片制备的变化。我将通过测试NL 1是否在Schaffer侧支-CA 1突触的NMDAR依赖性长时程增强（LTP）或长时程抑制（LTD）中起作用来跟踪这些实验。最后，为了评估NL 1对突触传递的控制是否具有突触前或突触后位点，我将使用出生后体内慢病毒注射来“拯救”具有点突变体的NL 1 KO小鼠中的个体细胞，所述点突变体破坏细胞外β-neurexin结合或细胞内与PSD-95的结合。这些实验的结果应该都恭维我们目前在体外知识，同时提高我们的理解的功能NL 1在成熟的海马电路。自闭症谱系障碍（ASD）包括高度遗传的神经发育障碍的异质组。神经脂素1是一种在突触中发现的分子，与家族性ASD有关。该提案旨在更好地了解NL 1在成熟大脑中的功能，以便有朝一日可以改善该基因异常导致的疾病。