The role of TRIO signaling in neuronal development, synaptic function, and circuit connectivity

TRIO 信号传导在神经元发育、突触功能和电路连接中的作用

基本信息

批准号：
10415377
负责人：
JESSICA A CARDIN
金额：
$ 71.62万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-01 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10415377
关键词：
Address Affect Affinity Alleles Anatomy Animal Model Anxiety Arousal Axon Behavior Behavioral Binding Biochemical Biochemistry Biology Bipolar Disorder Brain Brain Diseases Brain Pathology Calcium Cell Surface Receptors Cells Clustered Regularly Interspaced Short Palindromic Repeats Co-Immunoprecipitations Complex Coupling Defect Dendrites Dendritic Spines Development Disease Electron Microscopy Electrophysiology (science)Excitatory Synapse Functional disorder Genes Genetic Goals Image In Vitro Individual Intervention LGI1 gene Label Link Maps Mass Spectrum Analysis Measures Mediating Modeling Motor Activity Motor Cortex Mus Mutant Strains Mice Mutation Neocortex Neural Cell Adhesion Molecule L1 Neurodevelopmental Disorder Neurons Neuropil Pathology Patients Phenotype Process Proteins Proteomics Recombinant Proteins Risk Role Schizophrenia Series Signal Pathway Signal Transduction Structure Symptoms Synapses Synaptic Transmission Synaptic plasticity System TRIO gene Testing Variant Viral Whole-Cell Recordings Work autism spectrum disorder awake base behavioral phenotyping biochemical model comparative excitatory neuron genetic manipulation genetic variant in vivo in vivo imaging insight interdisciplinary approach knock-down light microscopy loss of function motor deficit multimodality neuron development neuronal excitability neuropsychiatric disorder optogenetics schizophrenia risk social deficits synaptic function tool

项目摘要

Abstract Heterozygous loss-of-function (LOF) or damaging variants in the TRIO gene are associated with increased risk for schizophrenia and autism spectrum disorders. However, the functional role of TRIO in neuronal biology and circuit function are not well understood, which limits the advance of therapies for these disorders. TRIO acts downstream of cell surface receptors to control axon and dendrite pathfinding, synapse development, and synaptic transmission. Deletion of a single TRIO allele in mouse cortical excitatory neurons drives reductions in cortical neuropil and defects in dendrite and synapse development and function, yielding social and motor deficits and increased anxiety and compulsivity. However, the links between specific TRIO mutations and subsequent consequences for cortical function are unknown. Here, we will integrate a broad array of highly complementary, interdisciplinary approaches including genetics, biochemistry and proteomics, optogenetic analysis of synaptic function, and multimodal in vivo imaging of cortical network dynamics to address this question. Our first aim will identify the biochemical mechanisms by which TRIO regulates cortical neuron development. We identified several new candidate TRIO signaling partners (PDE4A5, L1CAM, and the LGI1/ADAM22/ADAM23 complex) and will elucidate how they interact with TRIO to regulate cortical neuron dendritic arbor, dendritic spine, and synapse development. We also generated CRISPR mice heterozygous for three disorder-related TRIO variants - K1431M (autism), K1918X (schizophrenia), M2145T (bipolar disorder) - that differentially impact TRIO’s biochemical activities and yield different anatomical and behavioral phenotypes. We will use mass spectrometry-based comparative proteomics to discover new signaling partners differentially impacted by these discrete TRIO alleles. Our second aim will determine how different TRIO variants impact neuronal connectivity and synaptic function. We will assess the consequences of our TRIO CRISPR variants for cortical neuron development by measuring how they impact axon, dendrite, and synapse development, synaptic transmission and plasticity. We will also use viral Cre-mediated sparse TRIO disruption and whole cell recordings to test which deficits reflect cell- autonomous versus network level effects. Our third aim will test how alterations in TRIO impact the functional organization of cortical networks in vivo, taking advantage of our recently developed strategies for combining single cell and mesoscopic imaging of GCaMP6-labeled neurons to measure circuit organization in awake, behaving mice. Our overall goal is to understand how altered TRIO function impacts neuronal function at the cellular, synaptic, and network levels, providing a broad framework for understanding how genetic dysregulation drives changes in behavior.

抽象的三重基因中的杂合丧失功能丧失（LOF）或破坏性变体与精神分裂症和自闭症谱系障碍的风险增加。但是，三重奏在神经元中的功能作用生物学和电路功能尚不清楚，这限制了这些疾病的疗法的进步。三重奏作用于细胞表面受体的下游，以控制轴突和树突途径，突触发展，和突触传输。小鼠皮质兴奋神经元中单个三重奏等位基因的删除驱动减少在皮质神经胶体以及树突和突触的发展和功能中的缺陷，产生社交和运动缺乏症，焦虑和强迫性增加。但是，特定的三人突变与随后对皮质功能的后果未知。在这里，我们将集成一系列高度完成的跨学科方法，包括遗传学，生物化学和蛋白质组学，光遗传学分析突触函数和皮质网络动力学的多模式在体内成像，以解决此问题问题。我们的第一个目标将确定三重奏调节皮质神经元发育的生化机制。我们确定了几个新的候选三重奏信号合作伙伴（PDE4A5，L1CAM和 LGI1/ADAM22/ADAM23复合物），并将阐明它们与三人的相互作用以调节皮质神经元树突状乔木，树突状脊柱和突触的发育。我们还产生了CRISPR小鼠的杂合子三种与疾病相关的三人变体-K1431M（自闭症），K1918X（精神分裂症），M2145T（双相情感障碍） - 这对三人的生化活动有所不同，并产生不同的解剖和行为表型。我们将使用基于质谱的比较蛋白质组学以不同的方式发现新的信号伴侣受这些离散三重奏等位基因的影响。我们的第二个目标将决定不同的三变体如何影响神经元连接性和突触功能。我们将通过测量来评估三重奏CRISPR变体对皮质神经元发育的后果它们如何影响轴突，树突和突触的发展，突触传播和可塑性。我们也会使用病毒CRE介导的稀疏三重奏破坏和全细胞记录来定义反映细胞自主与网络级别效应。我们的第三个目标将测试三重奏的变化如何影响体内皮质网络的功能组织，利用我们最近开发的策略来结合单细胞和介观像的成像 GCAMP6标记的神经元在醒着中测量电路组织，行为小鼠。我们的总体目标是了解三重奏函数如何影响细胞，突触和网络水平上的神经元功能，提供一个广泛的框架来了解遗传失调如何导致行为变化。