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.

项目摘要突触是神经系统中沟通的基本单位，那里很大结构和功能的多样性有助于调整和校准信息传输。缺陷突触正确多元化的能力有助于多种多样的病因神经发育，精神病和神经退行性疾病。一种产生的方法突触功能的多样性是通过分子异质性，其中不同的组合基因在单个突触上表达，以实现特定的功能特性。但是，它越来越清楚，尽管难以解决，但出色的突触多样性可以可以从有限的分子机械组中实现。原则上，果蝇神经肌肉交界处（NMJ）是一个独特的强大模型，用于解决如何产生突触多样性复杂的遗传，电生理和成像方法。在这个系统中，两个独特的运动神经元会融合到单个肌肉靶标，在其中传播从强大而弱的输入一起驱动电机电路中的肌肉收缩。但是，一个无法选择性地隔离任何一个输入已成为主要限制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肉毒杆菌神经毒素（BONT）在强或弱突触下有选择地静音输入。初步数据表明，虽然每个神经元主要由相同在活动区域（一个核心组件）处的分子机械，以前认为起作用在所有活动区域上，实际上在强度与弱的角色上差异很大突触。我们将使用BONT沉默，超级分辨率成像和最新的钙记者针对释放站点以阐明活动区域纳米结构和功能的差异在强壮和弱突触之间。我们还将利用CRISPR的新创新诱变，以剖析八个核心活性区成分的专业功能 vs弱突触。最后，我们将询问这些核心活动区组件如何独特针对在强度与弱突触的调制和重塑的目标稳态突触可塑性。这些方法共同将基本见解解锁到谷氨酸能突触多样性如何通过可塑性建立和自适应修饰。最终，这种理解将阐明关键机制，通过这些机制有限的分子工具包可以实现谷氨酸能释放位点的功能性能。