Functional complexomics associated with maturation and activity-dependent plasticity of excitatory synapse

与兴奋性突触的成熟和活动依赖性可塑性相关的功能复杂组学

• 批准号: 537196039
• 负责人: Professor Dr. Bernd Fakler
• 金额: --
• 依托单位: Lehrstuhl für Physiologie II - Molecular Physiology
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/537196039?language=en
• 关键词: Functional; complexomics; associated; maturation; activity

Processing and storage of information in the brain fundamentally rely on proper signal transduction and activity-dependent dynamics in excitatory synapses. Key players in these synapses are AMPA-type glutamate receptors (AMPARs), macro-molecular complexes that drive almost any aspect of synapse physiology from synaptogenesis to electrical signal transduction and synaptic plasticity underlying memory formation and learning. We have recently uncovered biogenesis of AMPARs in the ER as a ‘multi-state assembly line’ and found that its impairment/disruption leads to severe consequences in both humans and rodents. In humans loss-of-function mutations in protein FRRS1l, a key determinant of the AMPAR assembly process, lead to severe forms of intellectual disability with strongest impairment in memory formation, motor skills and cognition. In mice, knock-out of FRRS1l abolished activity-dependent synaptic plasticity, reduced synapse formation and maturation and profoundly impaired learning. Interestingly, virally-driven re-expression of FRRS1l fully reversed all knock-out induced phenotypes and thus provided an experimental tool for ‘switching on’ formation of synapses and plastic behavior at will. In this project, we will use these latest insights for a first-time unbiased and comprehensive investigation of proteins that are required for building functional synapses with activity-driven plasticity. For this purpose we will (i) perform quantitative proteomic analyses on defined brain regions from FRRS1l knock-out mice before and after switching on AMPAR biogenesis by stereotactically delivered viruses, (ii) investigate identified key proteins and protein complexes for their subcellular distribution and dynamics and (iii) study their functional significance and characteristics in-vitro and in-vivo. Together, these analyses will decipher the molecular processes driving formation of excitatory synapses and their activity-dependent plasticity.

大脑中信息的处理和存储依赖于这些突触中的主要信号转导和活动的动力学。和学习。在多状态组装线中的AMPAR'并发现损伤/破坏会导致蛋白质FRS1L的人体丧失的两个人类的严重后果记忆形成，运动技能和认知的损害，FRRS1L的敲除依赖活动的突触塑料，降低突触形成和成熟和深刻的障碍。在此项目中，我们将使用这些最新见解的实验工具形成“形成”的突触和塑性行为。为此，我们将（i）通过立体固定传递的病毒栅极鉴定的关键蛋白质和蛋白质对其亚细胞分布和动力学研究他们的蛋白质和（iii）研究，对他们的亚电蛋白和蛋白质进行了定量的蛋白质组学分析对FRS1L敲除小鼠的定义脑部进行定量的蛋白质组学分析。功能显着性和C. haracteristics在体外和体内。