Synaptic Analysis of Neuroligin1 function

描述(由候选人提供)：这项建议的首要目标是探索神经连接蛋白的成熟突触功能，神经连接蛋白是一种突触特异性蛋白，与几种自闭症谱系障碍(ASD)有关。分子生物学和小鼠遗传学将与突触生理学相结合，以更好地了解Nl1的正常生物学功能，以及该基因突变对神经精神障碍中突触功能异常的可能贡献。除了为ASD的研究提供潜在的模型系统外，这项工作的结果还将有助于对成熟突触中NL功能的有限了解。通过这样做，他们可能会为未来精神障碍的分子干预提供新的靶点。之前对Nl1基因敲除小鼠的海马区的研究表明，Schaffer侧支/CA1突触的NMDAR/AMPAR比率降低了50%。我的前两个具体目标将通过分别描述对照和NL1突变急性海马片制备中AMPAR和NMDAR介导的电流的变化来具体说明这一观察结果。在这些实验之后，我将测试Nl1是否在Schaffer侧支-CA1突触的NMDAR依赖的长时程增强(LTP)或长时程抑制(LTD)中发挥作用。最后，为了评估NL1对突触传递的控制是否具有突触前或突触后的位置，我将使用出生后体内注射慢病毒来“拯救”具有点突变的NL1 KO小鼠的单个细胞，这些突变破坏了PSD-95的细胞外结合或细胞内结合。这些实验的结果应该既补充了我们目前的体外知识，又增强了我们对NL1在成熟的海马区回路中的功能的理解。自闭症谱系障碍(ASD)包括一组高度可遗传的神经发育障碍。神经连接素是一种在突触中发现的分子，与家族性自闭症有关。这项建议旨在更好地了解Nl1在成熟大脑中的功能，以便有一天可以改善该基因异常导致的疾病。

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.