Analysis of Ca2+ -Triggered Neurotransmitter Release

Ca2 触发神经递质释放的分析

基本信息

批准号：
7640959
负责人：
CHRISTIAN ROSENMUND
金额：
$ 29.6万
依托单位：
BAYLOR COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-07-01 至 2010-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7640959
关键词：
Action Potentials Address Affinity Attention Autistic Disorder Binding Biochemical Brain C2 Domain Cell membrane Communication Complex Dependency Disease Dominant-Negative Mutation Family member Finding by Cause Generic Drugs Genetic Goals In Vitro Individual Knockout Mice Light Mediating Membrane Membrane Fusion Molecular Mus Mutate Mutation N-terminal Neurons Neurotransmitters Pattern Penetration Phenotype Phospholipids Physiological Process Property Protein Family Proteins Reaction Research Personnel Role SNAP receptor Schizophrenia Screening procedure Shapes Signal Transduction Speed Structure-Activity Relationship Synapses Synaptic Transmission Tertiary Protein Structure Testing Time Transducers Translating Vesicle Viral base complement C2a complement C2b coping deficit syndrome design gain of function insight loss of function millisecond mutant neurotransmission neurotransmitter release overexpression programs protein complex reconstitution research study response synaptotagmin synaptotagmin I synaptotagmin VII

Action Potentials Address Affinity Attention Autistic Disorder Binding Biochemical Brain C2 Domain Cell membrane Communication Complex Dependency Disease Dominant-Negative Mutation Family member Finding by Cause Generic Drugs Genetic Goals In Vitro Individual Knockout Mice Light Mediating Membrane Membrane Fusion Molecular Mus Mutate Mutation N-terminal Neurons Neurotransmitters Pattern Penetration Phenotype Phospholipids Physiological Process Property Protein Family Proteins Reaction Research Personnel Role SNAP receptor Schizophrenia Screening procedure Shapes Signal Transduction Speed Structure-Activity Relationship Synapses Synaptic Transmission Tertiary Protein Structure Testing Time Transducers Translating Vesicle Viral base complement C2a complement C2b coping deficit syndrome design gain of function insight loss of function millisecond mutant neurotransmission neurotransmitter release overexpression programs protein complex reconstitution research study response synaptotagmin synaptotagmin I synaptotagmin VII

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项目摘要

DESCRIPTION (provided by applicant): Efficient and rapid transduction mechanisms at synapses are required for coherent oscillations and synchronization of activity of brain circuits. The presynapse transduces within <1 ms action potentials into a Ca2+-triggered synchronous fusion of neurotransmitter containing vesicles. Two protein families, the Complexins and Synaptotagmins, are key players in tuning the generic SNARE protein-based membrane fusion apparatus into a high speed transducer. Both proteins are known to interact with the SNARE complex. The goal of this study is to understand how these proteins accomplish fast fusion, and to determine which interactions are relevant for their function. We hypothesize that Complexin and Synaptotagmin 1 act by reducing the energy barrier of the fusion reaction, but they accomplish this using different molecular mechanisms. Following binding to the SNARE complex, Complexins seem to cause stabilization of a profusion complex, or alternatively, serve as an adaptor to enable Synaptotagmin 1 binding to the SNARE complex. Synaptotagmin 1 may trigger fast neurotransmitter release by Ca2+-dependent binding of its C2A and C2B domains to the plasma membrane, and this membrane penetration determines fusion rates by destabilizing the profusion membrane complex. To decipher the function of Complexin and Synaptotagmin, we propose an integrated approach using electrophysiological, structural, and biochemical experiments. First, we will analyze synaptic properties in neurons derived from Complexin 1/2/Synaptotagmin 1 knockout mice. We will then explore the structure-function relationship of protein domains that are putatively involved in synchronous release using a rescue approach. Finally, we will probe whether both proteins act independently or in conjunction with each other, and whether they act sequentially. A better understanding of the mechanism of synchronous release may help to find the cause and treatment of diseases that show disturbed oscillations and synchronization patterns in brain circuits, like Attention Deficit Syndrome, Autism, Huntingdon's Disease or Schizophrenia.

描述（由申请人提供）：突触时需要有效，快速的转导机制才能进行连贯的振荡和脑电路活性同步。 <1 ms的作用电位内的循环前传递到含有囊泡的神经递质的Ca2+触发同步融合中。两个蛋白质家族，即复合蛋白和突触量蛋白，是将一般的基因蛋白质膜融合设备调整为高速传感器的关键参与者。已知这两种蛋白质都与军鼓复合物相互作用。这项研究的目的是了解这些蛋白质如何完成快速融合，并确定哪些相互作用与其功能相关。我们假设复合蛋白和突触量1通过降低融合反应的能量屏障来起作用，但它们使用不同的分子机制来实现这一目标。在与军鼓复合物结合后，复合蛋白似乎会导致大量复合物的稳定，或者替代地作为适配器，使其能够与SNARE复合物结合突触。 SynaptoTagmin 1可能会触发Ca2+依赖性依赖性C2A和C2B结构域与质膜的快速神经递质释放，并且该膜穿透通过破坏膨胀膜复合物来确定融合速率。为了破译复合蛋白和突触毒素的功能，我们提出了一种使用电生理，结构和生化实验的综合方法。首先，我们将分析源自复合蛋白1/2/Synaptotagmin 1基因敲除小鼠的神经元的突触特性。然后，我们将探索使用救援方法推定参与同步释放的蛋白质结构域的结构功能关系。最后，我们将探测两种蛋白质是独立起作用还是相互作用，以及它们是否顺序起作用。更好地理解同步释放的机制可能有助于找到疾病的原因和治疗，这些疾病表现出干扰的振荡和同步模式，例如注意力缺陷综合征，自闭症，亨廷顿氏病或精神分裂症。