Isolation and Characterization of Synaptogenic Proteins

突触蛋白的分离和表征

• 批准号: 7796983
• 负责人: JUAN L BRUSES
• 金额: $ 21.25万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-12-01 至 2011-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7796983
• 关键词: Acetylcholine; Affect; Autistic Disorder; Binding; Biological Assay; Cell Adhesion Molecules; Cell Communication; Cell Membrane Proteins; Cell Surface Proteins; Cell membrane; Cell surface; Cells; Coculture Techniques; Collagen; Communication; Coupling; Development; Developmental Disabilities; Developmental Process; Elements; Embryonic Development; Endocytosis; Event; Extracellular Matrix; Extracellular Matrix Proteins; Extracellular Protein; Family; Fibroblast Growth Factor; Foxes; Future; Gene Expression; Gene Proteins; Genes; Goals; Immunoglobulins; In Vitro; Inositol; Laminin; Lead; Leucine-Rich Repeat; Ligands; Link; Membrane; Molecular; Muscle; Nerve; Nervous System Physiology; Neurons; Neurotransmitter Receptor; Nicotinic Receptors; Phase; Polystyrenes; Presynaptic Terminals; Proteins; Psyche structure; Research Project Grants; Schizophrenia; Site; Structure of ciliary ganglion; Surface; Synapses; Synaptic Membranes; Testing; Therapeutic; Therapeutic Intervention; Transcript; combinatorial; genome wide association study; genome-wide; genome-wide analysis; in vivo; membrane activity; nervous system disorder; postsynaptic; presynaptic; programs; protein expression; public health relevance; single molecule; synaptogenesis

DESCRIPTION (provided by applicant): The long-term goal of this research project is to elucidate the molecular mechanisms that regulate the formation of synaptic contacts. Synapses are a central component in the formation and the functioning of the nervous system and become affected in a variety of mental and neurological disorders. Therefore the discovery of the molecules and mechanisms that participate in the establishment of synaptic connections will contribute to our understanding of the formation of neuronal circuits, the causes of mental and neurological disorders, and may identify potential targets for therapeutic interventions. Synapse formation is a well-programmed developmental process that involves a variety of cell-cell interactions carried out by distinct groups of molecules, which are required for the recognition of a postsynaptic target, the stabilization of the contact between synaptic membranes, and the functional coupling between synaptic compartments. The present proposal focuses on the identification and characterization of proteins capable of inducing the formation of synaptic contacts between neurons during embryogenesis. Our central hypothesis is that the expression of membrane tethered or secreted ligands by the presumptive postsynaptic neuron are required to induce the differentiation of the presynaptic terminal and the establishment of a synaptic contact. For this reason, we carried out a genome-wide search for gene-transcripts that become expressed in postsynaptic neurons during the different phases of synapse development. This analysis led to the identification of a group of proteins containing immunoglobulin (Ig) domains, leucine-rich repeats (LRR), or other protein interacting domains that are substantially up-regulated during the initiation phase of synapse formation, and which have the molecular features of cell surface ligands suggesting that these proteins may possess synaptogenic activity. In this two-year project we propose 1) to test experimentally the synaptogenic activity of this subset of proteins up-regulated during the initiation of synapse formation, 2) to examine whether their combinatorial expression is needed to trigger synapse formation, and 3) to test whether cell surface expression of GPI- linked cell adhesion molecules regulates the surface expression levels of nicotinic acetylcholine neurotransmitter receptors. To this aim, in vitro cell assays will be used to determine the synaptogenic activity of these proteins by evaluating presynaptic terminal differentiation and cell surface protein expression levels. The identification of proteins which are sufficient to induce synapse formation in vitro will be further studied in the future to determine whether these proteins are necessary for the initiation of synapse formation in vivo and to examine the molecular mechanisms involved. PUBLIC HEALTH RELEVANCE: The long-term goal of this project is to elucidate the cellular and molecular mechanisms that participate in the assembly of synaptic contacts. The present proposal focuses on the identification and characterization of genes and proteins that participate in the initiation phase of synapse formation. The central hypothesis is that proteins expressed on the surface of presumptive postsynaptic neuron are necessary for inducing the differentiation of a presynaptic terminal at the site of contact. The importance of understanding the molecular mechanisms of synapse formation is underscored by the fact that synapses are the center piece for neuronal communication and become affected in a variety of mental and neurological disorders including, intellectual developmental disabilities, autism, and schizophrenia. Thus, the studies proposed in this application will lead to a better understanding of the molecular mechanisms that participate in the formation of synaptic connections, to the elucidation of the causes of mental and neurological disorders, and to the identification of therapeutic strategies.

描述（由申请人提供）：该研究项目的长期目标是阐明调节突触接触形成的分子机制。突触是神经系统形成和功能的核心组成部分，并受到各种心理和神经系统疾病的影响。因此，参与突触联系的分子和机制的发现将有助于我们理解神经元回路的形成，精神和神经系统疾病的原因，并可能识别出治疗干预措施的潜在目标。突触形成是一个良好编程的发育过程，涉及由不同的分子组进行的多种细胞 - 细胞相互作用，这是识别突触后靶标的所必需的，突触膜之间的接触稳定以及突触隔间之间的功能耦合。本提案的重点是鉴定和表征能够在胚胎发生过程中诱导神经元之间突触接触形成的蛋白质。我们的中心假设是，被推定的突触后神经元束缚或分泌的配体的表达需要诱导突触前末端的分化和建立突触接触。因此，我们在突触发育的不同阶段进行了全基因组搜索基因转录。该分析导致鉴定了一组含有免疫球蛋白（IG）结构域，富含亮氨酸的重复序列（LRR）或其他蛋白质相互作用域的蛋白质，这些蛋白质相互作用域在突触形成的起始阶段实质上被上调，并且在突触形成的起始阶段，这些蛋白具有这些蛋白质的分子特征，表明这些蛋白质可能具有Synaptodenatics的活性。在这个为期两年的项目中，我们建议1）通过实验测试突触形成过程中上调的蛋白质的突触生成活性，2）检查是否需要其组合表达来触发突触的形成，而3）以测试GPI-nected细胞粘附分子的细胞表面表达良好的表达蛋白氨基糖素。为此，通过评估突触前末端分化和细胞表面蛋白表达水平来确定这些蛋白质的突触性活性。将来将进一步研究足以在体外诱导突触形成的蛋白质的鉴定，以确定这些蛋白质是否对于启动体内突触形成是必要的，并检查涉及的分子机制。公共卫生相关性：该项目的长期目标是阐明参与突触接触组装的细胞和分子机制。目前的建议着重于参与突触形成起始阶段的基因和蛋白质的识别和表征。中心假设是在推定性突触后神经元表面表达的蛋白质对于诱导接触部位的突触前末端的分化是必要的。理解突触形成的分子机制的重要性是由突触是神经元交流的核心作品的事实，并受到了各种心理和神经系统疾病的影响，包括智力发育障碍，自闭症和精神分裂症。因此，本应用中提出的研究将使人们更好地理解参与突触连接形成的分子机制，阐明精神和神经系统疾病的原因以及鉴定治疗策略。