Persistent Pre- and Post-Synaptic Changes After Moderate Traumatic Brain Injury and Mitigation with MitoQ

中度创伤性脑损伤后持续的突触前和突触后变化以及 MitoQ 的缓解

• 批准号: 10643137
• 负责人: KEVIN Ka Wang WANG
• 金额: --
• 依托单位: VETERANS HEALTH ADMINISTRATION
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2028-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10643137
• 关键词: Activities of Daily Living; Affect; Afghanistan; Amygdaloid structure; Antioxidants; Anxiety; Area; Attenuated; Behavioral; Binding; Brain; Brain region; Calcium; Calmodulin; Caring; Chronic; Chronic Phase; Closed head injuries; Corpus striatum structure; DLG4 gene; Data; Diagnosis; Dopamine D1 Receptor; Dopamine D2 Receptor; Dopamine Receptor; Down-Regulation; Dyes; Enzyme-Linked Immunosorbent Assay; Face; Functional disorder; GABA-A Receptor; Glutamate Receptor; Glutamates; Healthcare; Hippocampus; Immunoelectron Microscopy; Injury; Iraq; Laboratories; Left; Lipids; Literature; Measures; Memory; Methods; Military Personnel; Mission; Mitochondria; Modeling; Modification; Morbidity - disease rate; Mus; Names; Neurotransmitters; Oral; Oral Examination; Outcome; Oxidative Stress Induction; Pilot Projects; Play; Postsynaptic Membrane; Prefrontal Cortex; Preparation; Presynaptic Terminals; Process; Proteins; Proteolysis; Publishing; Rain; Recovery; Research; Role; Scaffolding Protein; Sensory Receptors; Stains; Structural Protein; Structure; Synapses; Synaptosomes; Time; Traumatic Brain Injury; Veterans; blood-brain barrier crossing; cognitive function; controlled cortical impact; density; gephyrin; improved; injured; interest; mild traumatic brain injury; military veteran; mitoquinone; morris water maze; negative affect; neurobehavior; neurobehavioral; neurogranin; neurological rehabilitation; neurotransmission; neurotransmitter release; novel; novel therapeutics; oxidation; pharmacologic; postsynaptic; postsynaptic density protein; presynaptic; protein B; protein complex; receptor; receptor density

TBI was named the signature injury among military personnel served in the recent conflicts in Afghanistan and Iraq with more than 470,000 confirmed cases of TBI from 2010- 2020. In addition, more than 82,468 Veterans who use VA for their health care have been diagnosed with at least one TBI. Chronic synapse alternations represent an underappreciated area of interest in TBI pathobiology, with only a few published studies in the literature (1),(2),(3),(4) and even less examined in in the subacute/chronic phase of TBI. Our preliminary data show that there are in face persistent pre-synaptic and post-synaptic zone structural and functional vulnerability following moderate TBI in mice. These long-term changes could hinder synaptic adaptive mechanisms of the brain (synaptoplasticity) and therefore negatively affect brain recovery following TBI. We have evidence that presynaptic and post-synaptic protein and lipid components are especially vulnerable oxidative modifications and proteolysis, then leading to their down regulation. These synaptic changes, if persisted, could hinder synaptic adaptive mechanisms of the brain (synaptoplasticity) and therefore negatively affect brain recovery following TBI. Hypothesis: (i) TBI can cause sustained down-regulation of protein complexes at the pre-synaptic terminal active zone, which are associated with diminished vesicular neurotransmitter release function. (ii) In parallel, TBI also can cause oxidative and proteolytic damage of key protein components of the post-synaptic density (PSD) - PDZ-domain scaffold proteins (PSD93, PSD95, gephyrin) and associated calmodulin-regulator neurogranin, which can lead to instability & reduction of postsynaptic membrane-bound ionotropic glutamate receptor (NR2A, NR2B GluR1, GluR2), GABA receptor-A /B and dopamine receptors (D1, D2) and thus compromising the post-synaptic neurotransmission capacity. (iii) Studying TBI–induced pre- and post-synaptic protein alternations and dysfunctions can be facilitated by the use of synaptosome preparations isolated from injured mouse brain regions following moderate controlled cortical impact (CCI) and repeated close head injury (rCHI) are ideally suited to examine pre- and postsynaptic protein complex as well as in vesicular neurotransmitter release function and post-synaptic glutamate and dopamine receptor capacity. (iv) novel therapy with an oral mitochondria-targeting BBB-crossing antioxidant Mitoquinone (MitoQ) can help reduce post-TBI presynaptic and postsynaptic alterations and improve chronic neurobehavioral functions as supported by our pilot studies. In this proposal, we first aim to (1) study chronic post- CCI and post-rCHI alterations of key pre-synaptic and post-synaptic modulatory proteins in synaptosome/synaptoneurosome preparations and by immunohistochemical (IHC) staining and Immuno-electron microscopy. We then (2) examine chronic pre- synaptic vesicular neurotransmitter release capacity and post-synaptic glutamate receptors functional integrity and capacity in synaptosome preparations. We will (3) correlate changes of pre-synaptic and post-synaptic structures and functions in hippocampus, perirhinal cortex, prefrontal cortex and amygdala with corresponding changes in neurobehavioral endpoints involving these brain regions at 3 and 12 mo. post- TBI. (4) Examine the effects of post-TBI (CCI or rCHI) oral daily MitoQ treatment for 3 and 12 mo. on reducing oxidative stress induced damage to pre-synaptic and post-synaptic structures /functions, and improved synaptic mitochondrial function in hippocampus, perirhinal cortex, prefrontal cortex and amygdala with corresponding improvement in neurobehavioral endpoints involving these brain regions. In terms of matching US VA research and Veteran care missions, this proposal is directly relevant to US Veterans with chronic TBI who might have persistent pre- and post-synaptic changes that negatively affect their recovery and neurorehabilitation process.

在最近在阿富汗的冲突中，TBI被任命为军事人员的签名伤害 从2010年至2020年，伊拉克拥有超过470,000例TBI案件。此外，超过 82,468名将VA用于医疗保健的退伍军人被诊断出至少一名TBI。慢性突触 替代方案代表了TBI病理生物学感兴趣的领域，只有少数已发表的研究 在文献（1），（2），（3），（4）中，在TBI的亚急性/慢性相中检查的较少。我们的初步 数据表明，面部有持续的突触前和突触后区域结构和功能 小鼠中等TBI后的脆弱性。这些长期变化可能会阻碍突触自适应 大脑的机制（突触塑性），因此在TBI后对大脑恢复产生负面影响。我们 有证据表明突触前和突触后蛋白和脂质成分特别脆弱 氧化修饰和蛋白水解，然后导致其下降调节。这些综合变化，如果 坚持不懈，可能会阻碍大脑的突触自适应机制（突触塑性），因此负面 TBI后影响大脑恢复。假设：（i）TBI会导致蛋白质的持续下调 突触前末端活性区的复合物，该区域与囊泡减少有关 神经递质释放功能。 （ii）并行，TBI还可以引起钥匙的氧化和蛋白水解损伤 突触后密度（PSD）的蛋白质成分 - PDZ域支架蛋白（PSD93，PSD95， gephyrin）和相关的钙调蛋白 - 调节剂神经素蛋白，这可能导致不稳定和减少 突触后膜结合的离子型谷氨酸受体（NR2A，NR2B GLUR1，GLUR2），GABA受体-A /b和多巴胺受体（D1，D2），因此损害了突触后神经传递能力。 （iii） 可以通过使用TBI诱导的诱导前和突触后蛋白质替代方案和功能障碍。 中度受控皮质后，从受伤的小鼠脑区域分离出的突触体制剂 撞击（CCI）和重复的近距损伤（RCHI）非常适合检查突触前和突触后蛋白 复合物以及囊泡神经递质释放功能以及突触后谷氨酸和多巴胺 受体能力。 （iv）口服线粒体靶向BBB的抗氧化剂线喹酮的新疗法 （Mitoq）可以帮助减少TBI后突触前和突触后改变并改善慢性神经行为 正如我们的试点研究所支持的功能。 在此提案中，我们首先要（1）研究关键突触前的慢性CCI和RCHI后改变和RCHI的改变 突触体/突触体制剂中的突触后调节蛋白，并通过 免疫组织化学（IHC）染色和免疫电子显微镜。然后，我们（2）检查慢性预性 突触囊泡神经递质释放能力和突触后谷氨酸受体功能完整性 和突触体制剂的容量。我们将（3）相关联突触前和突触后的变化 海马，周围皮层，前额叶皮层和杏仁核的结构和功能与相应 涉及这些大脑区域3和12 mo的神经行为终点的变化。 tbi。 （4）检查 TBI（CCI或RCHI）口服每日MITOQ治疗3和12 mo的影响。减少氧化应激诱导的 损害突触前和突触后结构 /功能，并改善突触线粒体功能 海马，周围皮层，前额叶皮层和杏仁核，相应改善 神经行为终点涉及这些大脑区域。在与美国VA研究和退伍军人相匹配方面 护理任务，该提案与慢性TBI的美国退伍军人直接相关 以及突触后变化，对其康复和神经康复过程产生负面影响。