The molecular architecture of perineuronal nets

神经周围网络的分子结构

基本信息

批准号：
10625443
负责人：
Samuel Bouyain
金额：
$ 40.66万
依托单位：
UPSTATE MEDICAL 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：
2021-08-01 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10625443
关键词：
Architecture Binding Binding Proteins Biochemical Biological Assay CSPG3 gene Carbohydrates Cell Adhesion Molecules Cell Surface Proteins Cell Surface Receptors Cell surface Central Nervous System Chondroitin Sulfate Proteoglycan Complex Data Development Disease Event Exhibits Extracellular Matrix Extracellular Matrix Proteins Glycosaminoglycans Goals Health Homologous Gene Hyaluronan In Vitro Label Learning Life Ligands Link Macromolecular Complexes Memory Methodology Molecular Molecular Structure Neuronal Plasticity Neurons Neurophysiology - biologic function Pathogenesis Physiological Play Population Protein Tyrosine Phosphatase Proteins Reagent Recovery Role Specificity Stimulus Structure Surface Synapses Techniques Therapeutic Work X-Ray Crystallography aggrecan brevican cognitive function complex R contactin design developmental plasticity experimental study grasp in vivo information processing innovation insight interest janusin link protein macromolecular assembly nerve injury nervous system disorder neural neurodevelopment novel protein protein interaction receptor receptor binding receptor protein tyrosine phosphatase-type R response synaptogenesis tool versican

项目摘要

Perineuronal nets (PNNs) are conspicuous neural extracellular matrix (ECM) structures that have garnered significant interest over the last decade for the critical roles they play in neural developmental plasticity. These complex macromolecular structures are implicated in an array of cognitive functions, and are altered in a variety of neurological disorders. Despite the growing interest in PNN functions, the mechanisms by which they modulate neural functions are poorly understood, because there are currently no tools or techniques to manipulate PNNs specifically. We surmise that our inability to target and disrupt PNNs is primarily driven by a lack of understanding of their molecular composition or structure. Our goal in this proposal is to conduct a structure-function analysis of known PNN components as well as to identify proteins that anchor nets to neuronal surfaces. Using a powerful combination of in vitro and in vivo approaches, we have obtained strong preliminary data detailing how the newly identified PNN component receptor protein tyrosine phosphatase zeta (RPTPζ) associates with tenascin-R (TNR) within PNNs at a molecular level. Furthermore, our data indicate that the RPTPζ•TNR complex anchors PNNs to the neuronal cell surface via the GPI-linked protein contactin-1 (CNTN1), which makes CNTN1 the first surface binding protein for PNNs ever identified. Our central hypothesis is that there are a set of unique components and receptors of PNNs that nucleate PNNs and anchor them to specific neuronal cell surfaces, thereby defining their unique structure and functions. The overall objective of this proposal is to identify PNN-specific components and dissect the formation of PNNs through a unique combination of proximity-labeling assays, protein-binding assays, and protein X-ray crystallography in order to create the tools to target and manipulate these structures specifically and precisely. Our long-term goal is then to use these tools to dissect PNN function in order to better understand disease pathogenesis and ultimately to target PNNs therapeutically. Guided by our strong preliminary data, this proposal seeks to discover the unique components that guide the assembly of PNNs by pursuing three non-overlapping specific aims: 1) defining the role of the RPTPζ•TNR complex in anchoring PNNs to neuronal surfaces; 2) pursuing the biochemical and structural characterization of interactions between ACAN, HAPLN1, and TNR; and 3) identifying cell surface receptors and novel components of PNNs. The proposed work is significant because it will attempt to identify the key unique components that contribute to the formation and thereby function of PNNs. Successful completion of the aims will provide key insights and reagents to manipulate PNNs specifically and precisely and ultimately understand their functional mechanisms. This approach is innovative because it brings together a novel combination of physiological, biochemical and structural approaches to investigate these important macromolecular assemblies in the central nervous system. Ultimately, the proposed work could be transformative for the field and lead to key mechanistic insights into of PNN function in health and disease.

会内神经元网（PNN）是已获得的显着神经细胞外基质（ECM）结构在过去的十年中，对于神经发育塑性的批判性角色，这是对批判性角色的重大兴趣复杂的大分子结构与我有关的一系列认知功能，并在各种尽管对PNN功能的兴趣越来越大，但它们模拟神经功能是糟糕的联盟理解的，因为目前没有工具或技术专门操作PNN。缺乏对他们的分子组成或结构的理解。已知PNN成分的结构 - 功能分析，以鉴定纳入神经元的蛋白质表面使用体外和体内方法的强大组合数据详细介绍了新鉴定的PNN成分受体蛋白磷酸酶Zeta（RPTPζ）如何在分子水平的PNN中与Tenascin-R（TNR）相关联。 RPTPζ•TNRCompζ通过GPI连接的蛋白接触蛋白1（CNTN1），将PNN锚定在神经元细胞表面，使CNTN1成为曾经鉴定出的PNN的第一个表面结合蛋白。 PNN的atet成分和受体有核PNN并将其锚定在特定的神经元细胞表面，从而限制其独特的结构和功能。是识别PNN特异性成分和疾病，通过独特的组合形成PNN 接近标记的测定，蛋白质结合测定和蛋白质X射线晶体学，以创建工具针对和操纵这些结构。剖析PNN功能以更好地理解疾病发病机理并最终靶向PNN 在我们的强大初步数据的指导下，该提议旨在发现独特的组成部分通过追求三个非重叠的特定目的，指导PNN的组装：1）定义角色 RPTPζ•将PNN锚定在神经元表面的tnr复合物； 2） ACAN，HAPLN1和TNR之间的相互作用的表征； 3）识别细胞表面受体和 PNN的新颖组成部分。有助于PNN的形成和促进功能的组件。将提供关键的见解和试剂，以专门，精确地操纵PNN 它们的功能机制是创新的生理，生化和结构方法研究重要的大分子组件在中央神经系统中，最终的支撑作品可能是对该领域的变革对健康和疾病中PNN功能的机理见解。