Effects of ICP elevation on NeuN Negative Membrane Disrupted Neurons

ICP 升高对 NeuN 负膜破坏神经元的影响

• 批准号: 10541332
• 负责人: Martina Laura Hernandez
• 金额: $ 4.08万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10541332
• 关键词: Acute; Address; Affect; Animals; Anti-Inflammatory Agents; Automobile Driving; Award; Biological Assay; Calcitonin Gene-Related Peptide; Cathepsins B; Cell Death; Cell membrane; Cells; Cessation of life; Chronic; Clinical; Collaborations; Cytosol; Data; Dextrans; Diffuse; Electrophysiology (science); Etiology; Goals; Headache; Healthcare; Histologic; Hour; Impairment; Infusion procedures; Injury; Intervention; Intracranial Hypertension; Intracranial Pressure; Laboratories; Lead; Learning; Light; Lysosomes; Mediating; Mediator of activation protein; Membrane; Mentors; Microscopic; Migraine; Modeling; Molecular; Morbidity - disease rate; Nausea; Neuronal Injury; Neurons; Neurosciences; Outcome; Pathologic; Pathology; Peptide Hydrolases; Peptide antibodies; Persons; Pharmacologic Substance; Phase; Phenotype; Play; Population; Post-Traumatic Headaches; Protocols documentation; Publications; Rattus; Research; Research Personnel; Research Project Grants; Role; Side; Techniques; Therapeutic; Tracer; Training; Transcript; Traumatic Brain Injury; United States; Work; antagonist; burden of illness; career; cell injury; disability; experience; fluid percussion injury; in vivo; inhibitor; insight; novel; pain behavior; post-doctoral training; pre-clinical research; pre-doctoral; sound; triptans

Diffuse traumatic brain injury (TBI) is associated with various pathologies that lead to long-term impairments, including post-traumatic headache (PTH), particularly migraine. There are worse outcomes of TBI when compounded with elevated intracranial pressure (ICP). The objective of the F99 work is to elucidate the mechanism(s) that may be attributed to TBI-induced pathologies and ICP-mediated diffuse pathologies. The objective of the K00 is to investigate cellular and molecular mechanisms of migraine to become an investigator of PTH in TBI. One pathology, neuronal membrane disruption, has been shown to be induced acutely post-TBI. However, my initial studies present that membrane disruption in neurons can last beyond the acute timeframe and last for weeks post-TBI. Furthermore, data indicate that there is a subpopulation of cortical neurons that do not express NeuN and are membrane disrupted. With other injuries, a NeuN negative (NeuN-) presentation has been indicative of a reversion to an immature neuronal phenotype. If injured neurons are reverting to an immature phenotype, then this could be a compensatory mechanism, so identifying these neurons that also are membrane disrupted may provide more insight into the molecular mechanism of membrane disruption. Aim 1a) is to investigate the identity of NeuN- subpopulation of membrane disrupted neurons using intracerebroventricular (ICV) cell impermeable fluorescently-tagged dextran tracer in sham and central fluid percussion injured (CFPI) animals. Then using histological and molecular paradigms, I will evaluate membrane disrupted neurons for cellular NeuN expression, and expression of immature neuronal markers. Incidentally, the mechanism behind neuronal membrane disruption is unknown, and previous work from our lab shows that when TBI is compounded with an elevation in ICP in rats, that neuronal membrane disruption is increased in direct relation to the ICP elevation. Aim 1b) is to evaluate the effects of secondary ICP elevation on the NeuN- membrane disrupted subpopulation. I propose that lysosomal Cathepsin B (Cath B) is a potential mediator of membrane disruption, as previous findings reveal that Cath B re-localizes from the lysosome to the cytosol, which has been shown by other groups to initiate cell damage/death. I intend to evaluate the effects of secondary ICP elevation on the NeuN- membrane disrupted population using sham, CFPI animals and CFPI+elevated ICP animals with the same dextran protocol aforementioned, via microscopic and molecular approaches. Simultaneously, I will investigate the role of Cath B in vivo by inhibiting Cath B following sham and injury then using activity assays to verify inhibition as well as microscopic studies to evaluate the re-localization of Cath B. I expect in the F99 project that diffuse TBI paired with elevated ICP will see increases in membrane disrupted population later. Yet, a reduction in the compensatory NeuN- subpopulation as these neurons will endure a secondary insult. Finally, it is expected that Cath-B is re-localizing from the lysosome to the cytosol after TBI in membrane disrupted neurons and will be exacerbated with secondary ICP insult.

弥漫性创伤性脑损伤（TBI）与导致长期障碍的各种病理有关， 包括创伤后头痛（PTH），尤其是偏头痛。 TBI的结果更糟 与颅内压升高（ICP）复合。 F99工作的目的是阐明 可能归因于TBI诱导的病理和ICP介导的弥漫性病理的机制。这 K00的目的是研究偏头痛的细胞和分子机制成为研究者 TBI中的PTH。一种病理学是神经元膜破坏，已被证明是TBI急性引起的。 但是，我最初的研究表明，神经元的膜破坏可能持续超出急性时间表 并在TBI之后持续了几周。此外，数据表明，皮质神经元的亚群 没有表达neun，而膜被破坏。由于其他伤害，neun负（neun-）的表现具有 指示还原为未成熟的神经元表型。如果受伤的神经元恢复到未成熟 表型，那么这可能是一种补偿机制，因此识别这些神经元也是膜 被破坏可能会提供对膜破坏的分子机制的更多洞察力。目标1a）是 研究使用脑室内脑室中膜破坏神经元的Neun-Neun-亚群的身份 （ICV）在假和中央液体打击乐器中，细胞不渗透荧光标记的右旋晶状体示踪剂（CFPI） 动物。然后使用组织学和分子范例，我将评估膜破坏神经元的 细胞Neun表达和未成熟神经元标记的表达。顺便说一句，背后的机制 神经元膜破坏是未知的，我们实验室的先前工作表明，当TBI复合时 随着大鼠ICP升高，神经元膜破坏与ICP直接增加 海拔。 AIM 1B）是评估次级ICP高程对Neun-膜破坏的影响 亚种群。我建议溶酶体组织蛋白酶B（CATH B）是膜破坏的潜在介体， 如先前的发现表明，CATH B从溶酶体重新定位到细胞质，已通过 其他小组启动细胞损伤/死亡。我打算评估次级ICP高程对 Neun-膜使用假，CFPI动物和CFPI+升高的ICP动物中断了种群 通过显微镜和分子方法，上述相同的右旋体方案。同时，我会的 通过抑制假和伤害后的cath b，然后使用活动测定法调查CATA B在体内的作用 验证抑制作用以及显微镜研究，以评估CATH B的重新定位。 与ICP升高的TBI弥漫性TBI稍后会看到膜中断的人群中断。但是， 由于这些神经元将忍受二次损伤，补偿性新的亚群的减少。最后，它 预计CATH-B正在膜中破坏神经元的TBI后从溶酶体重新定位到细胞质 并将因次要ICP侮辱而加剧。