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.

TBI被命名为最近在阿富汗冲突中服役的军事人员的标志性伤害， 2010- 2020年，伊拉克有超过47万例TBI确诊病例。另外有超过 82，468名使用VA进行医疗保健的退伍军人被诊断出至少一次TBI。慢性突触 在创伤性脑损伤的病理生物学中，交替是一个未得到充分重视的领域，只有少数已发表的研究 在文献（1）、（2）、（3）、（4）中，甚至在TBI的亚急性/慢性期中更少地检查。我们的初步 结果表明，突触前和突触后存在着持续的结构和功能区 小鼠中度TBI后的脆弱性。这些长期的变化可能会阻碍突触的适应性 脑损伤的机制（突触可塑性），因此对脑损伤后的脑恢复产生负面影响。我们 有证据表明，突触前和突触后的蛋白质和脂质成分是特别脆弱的 氧化修饰和蛋白水解，然后导致其下调。这些突触变化，如果 持续存在，可能会阻碍大脑的突触适应机制（突触可塑性）， 影响脑外伤后的恢复假设：（i）TBI可引起蛋白质的持续下调 复合物在突触前终末活动区，这是与减少囊泡 神经递质释放功能(ii)同时，TBI还可引起关键细胞的氧化和蛋白水解损伤。 突触后密度（PSD）-PDZ-结构域支架蛋白（PSD 93，PSD 95， gephyrin）和相关的钙调素调节神经颗粒蛋白，这可能导致不稳定和减少 突触后膜结合离子型谷氨酸受体（NR 2A，NR 2B GluR 1，GluR 2），GABA受体-A /B和多巴胺受体（D1，D2），从而损害突触后神经传递能力。（三） 研究TBI诱导的突触前和突触后蛋白质改变和功能障碍可以通过使用 的突触体制备分离自损伤的小鼠脑区域， 撞击（CCI）和重复闭合性脑损伤（rCHI）非常适合于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和突触后谷氨酸和多巴胺 受体能力(iv)口服靶向血脑屏障交叉抗氧化剂米托醌的新疗法 （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 我们的试点研究支持的功能。 在这个提议中，我们首先旨在（1）研究CCI后和rCHI后关键突触前和突触后神经元的慢性改变， 突触体/突触神经体制剂中的突触后调节蛋白， 免疫组织化学（IHC）染色和免疫电镜。我们（2）检查慢性前， 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 和突触体制备的能力。我们将（3）将突触前和突触后的变化联系起来， 海马、嗅周皮层、前额叶皮层和杏仁核的结构和功能， 3个月和12个月时涉及这些脑区的神经行为终点的变化。创伤性脑损伤后。(4)检查 TBI（CCI或rCHI）后每日口服MitoQ治疗3个月和12个月的效果。减少氧化应激诱导的 对突触前和突触后结构/功能的损伤，以及改善突触线粒体功能， 海马、嗅周皮质、前额皮质和杏仁核， 涉及这些大脑区域的神经行为终点。在匹配美国退伍军人事务部研究和退伍军人方面， 护理任务，这项建议是直接相关的美国退伍军人与慢性TBI谁可能有持续的前 以及突触后变化，其负面地影响它们的恢复和神经康复过程。