Synaptic Vesicular Alterations after Traumatic Brain Injury

脑外伤后突触小泡的改变

• 批准号: 10683248
• 负责人: SHAUN CARLSON
• 金额: $ 42.48万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-15 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10683248
• 关键词: 3-Dimensional; Acute; Attention; Attenuated; Biological Assay; Blast Injuries; Brain; Brain Injuries; Cell membrane; Chronic; Clathrin; Clathrin Heavy Chains; Clathrin Light Chains; Cognition; Cognitive; Computer Models; Data; Development; Disease Pathway; Docking; Emotional; Endocytosis; Event; Female; Fluorescence; Functional disorder; Genes; Glutamates; Hippocampus; Homeostasis; Impaired cognition; Impairment; Individual; Injury; Knowledge; Laboratories; Learning; Light; Maps; Measures; Mediating; Membrane; Memory; Memory impairment; Microscopy; Modeling; Molecular; Neurons; Neurotransmitters; Pathology; Pathway interactions; Patients; Pharmacology; Play; Potassium; Proteins; Quality of life; Rattus; Recovery; Recovery of Function; Recycling; Reporting; Role; Severities; Sorting; Structure; Synapses; Synaptic Vesicles; Synaptosomes; System; Techniques; Testing; Therapeutic; Therapeutic Intervention; Traumatic Brain Injury; Vesicle; Viral; Viral Vector; Work; cognitive function; controlled cortical impact; density; gene network; genetic signature; immunoreactivity; improved; innovation; liquid chromatography mass spectrometry; male; neurobehavioral; neurotransmission; neurotransmitter release; new therapeutic target; novel; preclinical study; recruit; restoration; sex; single-cell RNA sequencing; spatial memory; synaptic function; targeted treatment; therapeutic target

Abstract Enduring cognitive, somatic, and emotional impairments, including difficulties with memory, attention, and learning, are reported as major contributors to reduced quality of life from patients living with a traumatic brain injury (TBI). Previous preclinical studies highlight the contribution of altered neurotransmission and synaptic dysfunction in the development of cognitive impairments after a TBI. We recently identified a novel synaptic pathology in which the density and distribution of the intrasynaptic vesicular pool is drastically reduced after an experimental TBI, with direct implications for impaired neurotransmission and cognition. A similar finding of altered vesicle docking was observed after low intensity blast injury, suggesting these alterations may occur across a broad spectrum of injury severity. Under normal conditions, neurotransmitter-containing vesicles are replenished and the vesicular pool is maintained through the primary recycling mechanism of clathrin-mediated endocytosis (CME). In the synapse, clathrin light chain (CLC) plays a central role in vesicular recycling through interaction with clathrin heavy chain and CME associated proteins for vesicle formation and replenishment of the vesicular pool. The effect of TBI on CLC and the contribution of CME deficits in synaptic dysfunction have not been examined. We provide preliminary data highlighting reductions in CLC and CME proteins for weeks post- injury in a rat model of controlled cortical impact (CCI). We also provide novel evidence of impaired hippocampal evoked neurotransmitter release, a functional readout of vesicular pool integrity and CME function. We hypothesize TBI-induced reductions in CME impairs the vesicular pool and neurotransmitter release, and that a targeted strategy to promote endocytosis improves neurotransmission and cognitive function after TBI. We propose utilizing innovative approaches to mechanistically interrogate the role of CLC and CME in post-traumatic synaptic dysfunction and investigate the functional effects of targeted therapeutic intervention on CLC abundance and CME function. In Aim 1, the effect of TBI on regional and sex-dependent changes in CLC and CME will be assessed in the early to subchronic recovery period following CCI. In Aim 2, we will examine the effect of a targeted strategy to increase CLC expression using neuron-specific adeno-associated viral modulation on CME function on protein readouts, evoked neurotransmission and cognitive function following CCI. Aim 3 will utilize an innovative and integrative Quantitative Systems Pharmacology approach to computationally model gene changes in network map pathways altered after TBI and to identify perturbed pathways responsive to AAV- CLC modulation after TBI. Successful completion of this work will bridge an important knowledge gap in understanding the detrimental effects of TBI on the synapse and identify CLC and CME function as a novel therapeutic target to promote functional recovery after a TBI.

抽象的 持久的认知，躯体和情绪障碍，包括在记忆，注意力和注意力方面遇到困难 据报道，学习是降低大脑外伤患者生活质量的主要贡献者 受伤（TBI）。先前的临床前研究突出了改变神经传递和突触的贡献 TBI后认知障碍发展的功能障碍。我们最近确定了一种新颖的突触 在一个病理学之后，在 实验性TBI，对神经传递和认知受损的影响直接影响。类似的发现 在低强度爆炸损伤后观察到囊泡对接改变，这表明可能发生这些改变 在广泛的伤害严重程度上。在正常情况下，含神经递质的囊泡是 补充并通过网格蛋白介导的主要回收机理来维持囊泡池 内吞作用（CME）。在突触中，网格蛋白轻链（CLC）在囊泡回收中起着核心作用 与网格蛋白重链和CME相关蛋白的相互作用，以形成囊泡和补充 囊泡池。 TBI对CLC的影响以及CME缺陷在突触功能障碍中的影响尚未 被检查。我们提供了初步数据，该数据突出了CLC和CME蛋白的减少数周 - 受控皮质影响（CCI）大鼠模型的损伤。我们还提供了海马受损的新证据 诱发神经递质释放，这是囊泡池完整性和CME函数的功能读数。我们 假设CME的TBI诱导的减少损害了囊泡池和神经递质的释放，并且A 促进内吞作用的有针对性策略可改善TBI后神经传递和认知功能。我们 提出利用创新方法来机械地询问CLC和CME在创伤后的作用 突触功能障碍并研究靶向治疗干预对CLC的功能作用 丰度和CME功能。在AIM 1中，TBI对CLC和性别依赖性变化的影响以及 CME将在CCI之后的早期至亚基恢复期进行评估。在AIM 2中，我们将检查 使用神经元特异性腺相关的病毒调节来增加CLC表达的靶向策略的影响 在CCI后，关于蛋白质读数的CME功能，诱发神经传递和认知功能。目标3意志 利用一种创新和综合定量系统药理学方法来计算模型 网络图路径的基因变化在TBI之后发生了变化，并确定对AAV-响应的扰动途径 TBI后的CLC调制。成功完成这项工作将弥合重要的知识差距 了解TBI对突触的有害影响，并识别CLC和CME的功能是一种新颖 治疗靶标可在TBI后促进功能恢复。