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.

摘要 持久的认知、躯体和情绪障碍，包括记忆、注意力和精神障碍 据报道，学习是导致脑外伤患者生活质量下降的主要原因 损伤(TBI)。先前的临床前研究强调了神经传递和突触改变的作用 脑外伤后认知功能障碍的发展。我们最近发现了一种新的突触 突触内水泡池的密度和分布在急性脑损伤后急剧减少的一种病理 实验性脑外伤，与神经传递和认知受损有直接关系。一项类似的发现 在低强度冲击伤后观察到改变的囊泡停靠，提示这些改变可能发生。 伤情严重程度各不相同。在正常情况下，含有神经递质的囊泡 通过网状蛋白介导的初级循环机制来补充和维持囊泡池 内吞作用(CME)。在突触中，网状蛋白轻链(clathrin light chain，CLC)通过 与胞蛋白重链和CME相关蛋白相互作用对囊泡形成和补充的影响 水泡泳池。颅脑损伤对CLC的影响以及CME缺陷在突触功能障碍中的作用 被检查过了。我们提供了初步数据，强调了CLC和CME蛋白在术后几周的减少。 控制皮质撞击(CCI)大鼠模型中的损伤。我们还提供了海马区受损的新证据。 诱发的神经递质释放，囊泡池完整性和CME功能的功能读数。我们 假设脑损伤诱导的CME减少会损害囊泡池和神经递质的释放，并且 促进内吞作用的靶向策略可改善脑外伤后的神经传递和认知功能。我们 建议利用创新的方法机械地审问《中图法》和《继续医学教育》在创伤后的作用 突触功能障碍与靶向治疗对慢性淋巴细胞性肺癌功能的影响 丰度与CME功能。在目标1中，脑创伤对CLC和CLC的区域性和性别依赖性变化的影响 CME将在CCI后早期到亚慢性恢复期进行评估。在目标2中，我们将研究 利用神经元特异腺相关病毒调节增加ClC表达的靶向策略的作用 持续脑电对CCI后蛋白质读数、诱发神经传递和认知功能的影响。目标3将 利用创新和综合的定量系统药理学方法建立计算模型 颅脑损伤后网络图通路的基因变化及识别对甲型肝炎病毒反应的扰动通路 脑损伤后的CLC调制。这项工作的成功完成将弥合以下方面的重要知识鸿沟 了解颅脑损伤对突触的不利影响并确定CLC和CME是一种新的功能 促进脑外伤后功能恢复的治疗目标。