Administrative Supplement (Diversity) to Generating functional diversity from molecular homogeneity at glutamatergic synapses

从谷氨酸能突触的分子同质性生成功能多样性的行政补充（多样性）

基本信息

批准号：
10841899
负责人：
DION KAI DICKMAN
金额：
$ 3.36万
依托单位：
UNIVERSITY OF SOUTHERN CALIFORNIA
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-07-01 至 2027-11-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10841899
关键词：
Action Potentials Address Administrative Supplement Alzheimer&apos s Disease Biological Models Botulinum Toxins Calcium Calibration Chemosensitization Clustered Regularly Interspaced Short Palindromic Repeats Communication Coupling Data Defect Development Disease Drosophila genus Electrophysiology (science)Epilepsy Etiology Failure Genes Genetic Glutamates Goals Health Heterogeneity Homologous Gene Individual Mammals Mental disorders Microscopy Modeling Molecular Molecular Machines Motor Motor Neurons Muscle Muscle Contraction Mutagenesis Nanostructures Nervous System Neurodegenerative Disorders Neurodevelopmental Disorder Neuromuscular Junction Neurons Neurophysiology - biologic function Orthologous Gene Phase Positioning Attribute Process Property Protein Isoforms RNA Splicing Reagent Reporter Role Schizophrenia Site Structure Synapses Synaptic plasticity System Toxic effect Vesicle autism spectrum disorder botulinum toxin type C experience experimental study fly imaging approach innovation insight nanoscale nervous system disorder neural circuit neuropsychiatric disorder neurotransmission neurotransmitter release postsynaptic presynaptic prevent ratiometric scaffold superresolution imaging synaptic function transmission process ultra high resolution

项目摘要

PROJECT SUMMARY Synapses are fundamental units of communication in the nervous system, where immense diversity in structure and function serve to tune and calibrate information transfer. Defects in the ability of synapses to properly diversify contribute to the etiology of a variety of neurodevelopmental, psychiatric, and neurodegenerative diseases. One means of generating diversity in synaptic function is through molecular heterogeneity, where combinations of distinct genes are expressed at individual synapses to enable specific functional properties. However, it has become increasingly clear, though difficult to resolve, that remarkable synaptic diversity can be achieved from a limited set of molecular machinery. In principle, the Drosophila neuromuscular junction (NMJ) is a uniquely powerful model to address how synaptic diversity is generated given the sophisticated genetic, electrophysiological, and imaging approaches. In this system, two distinct motor neurons converge to co-innervate individual muscle targets, where transmission from a strong and weak input together drive muscle contraction in the motor circuit. However, an inability to selectively isolate transmission from either input has been a major limitation towards understanding synaptic diversity in this system. Here, we propose to use expression of a unique Botulinum NeuroToxin (BoNT) to selectively silence transmission at strong or weak synaptic inputs. Preliminary data suggests that while each neuron is largely composed of the same molecular machinery at active zones, one core component, previously thought to function universally at all active zones, actually subserves dramatically different roles at strong vs weak synapses. We will use BoNT silencing, super resolution imaging, and the latest calcium reporters targeted to release sites to illuminate differences in active zone nanostructure and function between strong and weak synapses. We will also leverage new innovations in CRISPR mutagenesis to dissect the specialized functions of eight core active zone components at strong vs weak synapses. Finally, we will interrogate how these core active zone components are uniquely targeted for modulation and remodeling at strong vs weak synapses in the context of homeostatic synaptic plasticity. Together, these approaches will unlock fundamental insights into how glutamatergic synaptic diversity is established and adaptively modified through plasticity. Ultimately, this understanding will illuminate key mechanisms through which heterogeneous functional properties at glutamatergic release sites are enabled by a limited molecular toolkit.

项目概要突触是神经系统中沟通的基本单位，其中巨大的结构和功能的多样性有助于调整和校准信息传递。缺陷在突触适当多样化的能力有助于多种疾病的病因学神经发育、精神和神经退行性疾病。一种生成方式突触功能的多样性是通过分子异质性实现的，其中不同的组合基因在各个突触处表达以实现特定的功能特性。然而，它尽管难以解决，但已经变得越来越清楚，显着的突触多样性可以可以通过一组有限的分子机器来实现。原则上，果蝇神经肌肉 junction (NMJ) 是一个独特的强大模型，可以解决给定的情况下突触多样性是如何生成的复杂的遗传、电生理和成像方法。在这个系统中，两个不同的运动神经元聚集在一起共同神经支配各个肌肉目标，其中传输强输入和弱输入共同驱动运动回路中的肌肉收缩。然而，一个无法选择性地隔离来自任一输入的传输一直是对了解该系统中的突触多样性。在这里，我们建议使用独特的表达方式肉毒杆菌神经毒素 (BoNT) 可选择性地抑制强或弱突触的传递输入。初步数据表明，虽然每个神经元主要由相同的神经元组成活性区的分子机械，一个核心组成部分，以前被认为起作用在所有活跃区域普遍存在，实际上在强弱区域中发挥着截然不同的作用突触。我们将使用 BoNT 沉默、超分辨率成像和最新的钙报告基因针对释放位点以阐明活性区纳米结构和功能的差异强突触和弱突触之间。我们还将利用 CRISPR 的新创新诱变以强强度剖析八个核心活性区成分的专门功能与弱突触。最后，我们将询问这些核心活动区域组件是如何独特地针对强突触和弱突触的调制和重塑稳态突触可塑性。这些方法共同将解锁对以下问题的基本见解：谷氨酸突触多样性是如何通过可塑性建立和适应性修改的。最终，这种理解将阐明异质性的关键机制谷氨酸释放位点的功能特性是通过有限的分子工具包实现的。