Postsynaptic roles of ankyrins

锚蛋白的突触后作用

• 批准号: 10365120
• 负责人: Peter Penzes
• 金额: $ 63.98万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-03-01 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10365120
• 关键词: ANK2 gene; ANK3 gene; Ankyrin Repeat; Ankyrins; Axon; Behavior; Behavioral; Biochemical; Bioinformatics; Biological Process; Biology; Bipolar Disorder; Brain; Brain Diseases; Calcium; Cell membrane; Complex; Data; Dendrites; Dendritic Spines; Deubiquitination; Development; Family Study; Funding; Funding Opportunities; Future; GTP-Binding Proteins; Genome; Glutamates; Homer 1; Human; Hybrids; Image; In Vitro; Intellectual functioning disability; Knock-out; Knockout Mice; Laboratories; Maintenance; Methods; Molecular and Cellular Biology; Morphology; Mus; Mutation; Neuraxis; Neurobiology; Neurodevelopmental Disorder; Neurons; Pathogenicity; Phenotype; Plasmids; Play; Positioning Attribute; Process; Proteins; Proteomics; Reagent; Regulation; Reporter; Research; Resolution; Rodent; Role; Schizophrenia; Site; Structure; Susceptibility Gene; Synapses; Synaptosomes; Testing; Time; Transforming Growth Factor beta; United States National Institutes of Health; Variant; Vertebral column; Yeasts; autism spectrum disorder; cell type; conditional knockout; endocannabinoid signaling; exome sequencing; frontal lobe; genome-wide; in vivo; mouse model; neuroproteomics; neuropsychiatric disorder; novel; postsynaptic; rare variant; risk variant; scaffold; spatiotemporal; synaptic function; two photon microscopy

ABSTRACT Extensive evidence implicates glutamatergic synapses as key pathogenic sites in neuropsychiatric disorders (NPDs). Recently-emerging evidence has implicated ankyrin proteins, including Ankyrin-G (AnkG), encoded by the ANK3 gene, and Ankyrin-B (AnkB), encoded by ANK2, in the biology of central nervous system synapses. While conventionally AnkG and –B have been seen as playing roles mainly at the axon initial segment, their functions in central glutamatergic synapses have only begun to be investigated, including in studies initiated by our laboratory. During the previous funding period, we have made extensive progress in understanding the regulation and interactome of AnkG at synapses and in dendrites. Our preliminary findings broaden our understanding of the neurobiology of ankyrins, and implicate them in several novel process, including deubiquitination and endocannabinoid signaling. In this proposal we will use neuroproteomics, conditional knockout and reporter mice, super-resolution and in vivo two-photon microscopy, calcium imaging, bioinformatics, and behavioral analyses, to investigate the functions of AnkG's protein partners in dendrite and at glutamatergic synapses. We hypothesize that AnkG performs multiple novel functions including supramolecular scaffold complex maintenance and regulation of endocannabinoid signaling, through which it controls spiny synapse development, structure, function, as well as circuit level network activity and behavior, in a cell type and developmental period-specific manner. We will test this hypothesis in the following Aims: Aim 1. To investigate the impact of cell type-dependent and developmentally-timed knock out of AnkG on brain structure, function, and behavior across maturation. Aim 2. To investigate the impact of cell type-dependent and developmentally-timed knock out of AnkB on brain structure, function, and behavior across maturation. Aim 3. To determine the mechanisms and functional significance of the interaction of AnkG with DAGLĮ. The proposed studies are highly novel and impactful, given that the synaptic functions of ankyrins are only beginning to be investigated. This proposal will be the first to investigate the function of several recently-discovered AnkG-interacting proteins in unexpected biological processes, using multiple cutting edge methods, and will open new avenues for many future studies.

摘要 大量证据表明谷氨酸能突触是神经精神疾病的关键致病部位 (NPDS)。最近出现的证据表明，包括Ankyrin-G(AnkG)在内的Ankyrin蛋白， ANK3基因编码的Ankyrin-B和ANK2编码的Ankyrin-B(AnkB)在中枢神经生物学中 系统突触。传统上，AnkG和-B被认为主要在轴突起作用 最初的片段，它们在中枢谷氨酸能突触中的功能才刚刚开始研究， 包括我们实验室发起的研究。在上一次融资期间，我们取得了广泛的进展 AnkG在突触和树突中的调节和相互作用的研究进展。我们的 初步的发现加深了我们对Ankyrins神经生物学的理解，并将其与 一些新的过程，包括去泛素化和内源性大麻素信号。在本提案中，我们将 使用神经蛋白质组学、条件基因敲除和报告小鼠、超分辨率和体内双光子 显微镜、钙成像、生物信息学和行为分析，以研究 AnkG的蛋白质在树突和谷氨酸能突触中配对。我们假设AnkG执行 多种新功能，包括超分子支架复合体的维护和调节 内源性大麻素信号，通过它控制棘突触的发育、结构、功能，如 以及电路级网络活动和行为，以细胞类型和发育时期特定的方式。 我们将在以下目标中检验这一假说：目的1.调查细胞类型依赖的影响 AnkG基因敲除对大脑结构、功能和行为的影响 成熟。目的2.研究细胞类型依赖和发育时序基因敲除的影响 AnkB对大脑结构、功能和行为发育的影响。目标3.确定机制 以及AnkG与DAGLĮ相互作用的功能意义。拟议的研究是非常新颖的 考虑到对Anyrins的突触功能的研究才刚刚开始，这是很有影响力的。这 Proposal将首次研究最近发现的几种AnkG相互作用蛋白的功能 在意想不到的生物过程中，使用多种尖端方法，并将为 许多未来的研究。