Effects of Developmental Ethanol Exposure on Cerebellar Microglia and Purkinje Cells

发育期乙醇暴露对小脑小胶质细胞和浦肯野细胞的影响

• 批准号: 10533906
• 负责人: MaKenna Cealie
• 金额: $ 4.68万
• 依托单位: UNIVERSITY OF ROCHESTER
• 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 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10533906
• 关键词: ARHGEF5 gene; Acute; Adult; Affect; Age; Agonist; Alcohol consumption; Animals; Attenuated; Behavior; Behavioral; Birth; Brain; Cell Communication; Cells; Cerebellum; Chronic; Cognitive; Control Animal; Dendrites; Dendritic Spines; Development; Disease; Electron Microscopy; Ethanol; Evolution; Exposure to; Fetal Alcohol Spectrum Disorder; Human; Image; Immune; Immunohistochemistry; Impaired cognition; Impairment; Inflammation; Inflammatory; Inflammatory Response; Intervention; Knowledge; Learning; Life; Link; Long-Term Effects; Mediating; Microglia; Morphology; Mus; Neuraxis; Neuroimmune; Neurons; Output; PPAR alpha; PPAR gamma; Pathologic; Pathology; Phagocytosis; Phenotype; Pioglitazone; Play; Process; Psyche structure; Purkinje Cells; Quantitative Reverse Transcriptase PCR; Reporting; Rodent; Role; Shapes; Structure; Synapses; Testing; Third Pregnancy Trimester; Time; alcohol effect; alcohol exposure; behavioral impairment; cell motility; disability; emerging adult; experimental study; immune activation; in vivo; in vivo two-photon imaging; morphometry; motor impairment; mouse model; neuronal cell body; neuronal circuitry; postnatal; response; therapeutic target; time use; two-photon

Project Summary Fetal alcohol spectrum disorders (FASD) are the most common cause of non-heritable, preventable mental disability, occurring in almost 5% of births in the U.S. There is no known cure for FASD, and its mechanisms remain unclear. A wide range of cognitive, behavioral, and physical impairments have been reported in FASD, including deficits in behaviors related to the cerebellum. These changes in behavior may arise from ethanol's effects on the cellular level. The sole output of the cerebellum, Purkinje cells, as well as microglia, the immune cells of the Central Nervous System, are both impacted by developmental ethanol exposure. Reduced numbers of both neurons and microglia, as well as alterations in Purkinje cell excitability and firing have been reported. After developmental ethanol exposure, microglia display a phenotype associated with immune activation and release pro-inflammatory factors. Peroxisome proliferator-activated receptor-γ (PPAR-γ) agonists can block this immune activation and have been shown to attenuate some of the inflammatory responses in microglia and reduce Purkinje cell loss in rodents. This suggests that microglia may be a therapeutic target in FASD. Microglia are known to shape neuronal circuit development and connectivity in the cerebellum, which is linked to microglial structural dynamics. How ethanol affects these dynamics and how that impacts microglia- Purkinje cell interactions is unknown. Additionally, it is not yet known when these changes occur and how they are maintained or progressively altered into adulthood. Elucidating how ethanol-induced changes in microglia mediate some of the pathological changes in cerebellar Purkinje cells may be critical for understanding the onset of FASD pathology. Furthermore, modulating microglial survival and activity during ethanol exposure through a PPAR-γ agonist may provide some answers and potential therapies for these diseases. Thus, I hypothesize that ethanol induces neuroimmune changes in cerebellar microglia that alter their dynamics and interactions with Purkinje cells, and reducing microglia-mediated inflammation through a PPAR-γ agonist mitigates the pathological effects of ethanol. To test this hypothesis, I will pursue two aims using a mouse model of FASD. The first will investigate how developmental ethanol and a PPAR-γ agonist affects microglial phenotype over time using in vivo two-photon imaging of microglial dynamics, immunohistochemistry, and quantitative real time PCR. The second will determine if Purkinje cell and microglia interactions are affected throughout life by developmental ethanol exposure and PPAR-γ agonist administration with two-photon imaging, immunohistochemistry, and electron microscopy. These experiments will elucidate the effects of cerebellar microglia on Purkinje cells in the cerebellum after developmental ethanol exposure and assess microglia as a potential target to mitigate disease pathology in a mouse model of FASD.

项目摘要 胎儿酒精谱（FASD）是最常见的原因，可预防的 精神疾病，在美国近5％的出生中发生，没有已知的FASD治疗 机制仍然不清楚。多种认知，行为和身体障碍已经存在 在FASD中报告的，包括与小脑有关的行为定义。这些行为的变化可能会出现 从乙醇对细胞水平的影响。小脑，浦肯野细胞以及小胶质细胞的唯一输出 中枢神经系统的免疫细胞都受到发育性乙醇暴露的影响。 神经元和小胶质细胞的数量减少，以及Purkinje细胞兴奋性和解散的改变 据报道。在开发乙醇暴露后，小胶质细胞显示与免疫相关的表型 激活和释放促炎因子。过氧化物组增殖物激活的受体-γ（PPAR-γ）激动剂 可以阻止这种免疫激活，并已证明可以减弱一些炎症反应 小胶质细胞并减少啮齿动物中的浦肯野细胞损失。这表明小胶质细胞可能是 Fasd。已知小胶质细胞塑造小脑中神经元电路的发展和连通性，这是 与小胶质结构动力学相关。乙醇如何影响这些动态及其如何影响小胶质细胞 Purkinje细胞相互作用尚不清楚。此外，尚不清楚这些变化何时发生以及它们如何 维持或逐渐改变为成年。阐明乙醇诱导的小胶质细胞的变化 介导小脑浦肯野细胞的某些病理变化可能对于理解发作至关重要 FASD病理学。此外，通过A调节小胶质细胞生存和活性 PPAR-γ激动剂可以为这些疾病提供一些答案和潜在疗法。那我假设 乙醇诱导小脑小胶质细胞的神经免疫性变化，从而改变其动力学和与 Purkinje细胞，并通过PPAR-γ激动剂减少小胶质细胞介导的注射会减轻 乙醇的病理作用。为了检验这一假设，我将使用FASD的小鼠模型追求两个目标。 第一个将研究发育性乙醇和PPAR-γ激动剂如何影响小胶质表型 使用小胶质细胞动力学，免疫组织化学和定量实时的体内两光子成像的时间 pcr。第二个将确定Purkinje细胞和小胶质细胞相互作用是否会受到一生的影响 具有两光子成像的发育性乙醇暴露和PPAR-γ激动剂， 免疫组织化学和电子显微镜。这些实验将阐明小脑的影响 发育性乙醇暴露和评估小胶质细胞后小脑的Purkinje细胞上的小胶质细胞作为一种 在FASD小鼠模型中减轻疾病病理的潜在靶标。