Nanoscale cellular and molecular fingerprints of synaptic diversity

突触多样性的纳米级细胞和分子指纹

• 批准号: 391076133
• 负责人: Professor Dr. Stephan J. Sigrist
• 金额: --
• 依托单位: Institut für Biologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2017
• 资助国家: 德国
• 起止时间: 2016-12-31 至 2020-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/391076133?language=en
• 关键词: Nanoscale; cellular; molecular; fingerprints; synaptic

Functional synaptic diversity is critical for routing and encoding sensory information in the brain. Yet, the molecular and biophysical underpinnings of such diversity remain unexplored. We hypothesize that nanoscale variations in the number and location of active zone scaffold proteins, as well as synapse-specific expression of the molecular machinery controlling their fusion, contribute to functional synaptic diversity. Using the mouse somatosensory cortex and cerebellum we will identify the molecular and biophysical mechanisms underlying the diversity of inhibitory and excitatory synapses (strong and weak), which are responsible for sensory perception. Many candidate molecules influencing synaptic behavior have been identified in the drosophila model system. Here we propose to use super-resolution imaging of synaptic molecules to identify their binding partners and characterize their nanoscale architecture (molecular fingerprint), which ultimately influences the distance between calcium channels and synaptic vesicles. In order to establish a structure function relationship, we will estimate coupling distances using combining optical and biophysical approaches at single boutons. Finally, causal experiments will be performed by genetic alteration (Crisper/Cas9 gene editing) of key active zone scaffold proteins to confirm their specific role in setting coupling distance. We expect to elucidate general molecular rules defining functional diversity throughout the brain. Because of the involvement of presynaptic proteins in several brain disorders and the molecular nature and cross-brain breadth of our study, this research will provide a foundation for therapeutic future interventions.

功能性突触多样性对于大脑中的感觉信息的路由和编码至关重要。然而，这种多样性的分子和生物物理基础仍未得到探索。我们假设，活性区支架蛋白的数量和位置的纳米级变化，以及控制其融合的分子机制的突触特异性表达，有助于功能性突触多样性。使用小鼠体感皮层和小脑，我们将确定的分子和生物物理机制的多样性的抑制性和兴奋性突触（强和弱），这是负责感觉知觉。许多影响突触行为的候选分子已经在果蝇模型系统中被鉴定。在这里，我们建议使用突触分子的超分辨率成像来识别它们的结合伙伴，并表征它们的纳米级结构（分子指纹），这最终影响钙通道和突触囊泡之间的距离。为了建立一个结构功能关系，我们将估计耦合距离结合使用光学和生物物理方法在单个扣。最后，将通过关键活性区支架蛋白的遗传改变（Criminal/Cas9基因编辑）进行因果实验，以确认它们在设定偶联距离中的特定作用。我们希望阐明定义整个大脑功能多样性的一般分子规则。由于突触前蛋白参与了几种脑部疾病，以及我们研究的分子性质和跨脑广度，这项研究将为未来的治疗干预提供基础。