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: 10731361
• 负责人: MaKenna Cealie
• 金额: $ 4.77万
• 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/10731361
• 关键词: ARHGEF5 gene; Acute; Adult; Affect; Age; Agonist; Alcohol consumption; Animals; Attenuated; Behavior; Behavioral; Birth; Brain; Cell Communication; Cells; Central Nervous System; Cerebellum; Chronic; Cognitive; Control Animal; Dendrites; Development; Disease; Electron Microscopy; Ethanol; Evolution; 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; Neuroimmune; Neurons; Output; PPAR gamma; Pathologic; Pathology; Phagocytosis; Phenotype; Pioglitazone; Process; Psyche structure; Purkinje Cells; Reporting; Rodent; Role; Shapes; Structure; Synapses; Testing; Third Pregnancy Trimester; Time; Vertebral column; 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 胎儿酒精谱系障碍（FASD）是最常见的非遗传性，可预防的原因。 精神残疾，发生在近5%的出生在美国有没有已知的治愈FASD，其 其机制仍不清楚。广泛的认知，行为和身体障碍已经被 在FASD中报告，包括与小脑相关的行为缺陷。这些行为上的变化可能会出现 乙醇对细胞水平的影响。小脑浦肯野细胞和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期乙醇暴露的影响。 神经元和小胶质细胞数量的减少，以及浦肯野细胞兴奋性和放电的改变， 被举报。乙醇暴露后，小胶质细胞表现出与免疫相关的表型， 激活并释放促炎因子。过氧化物酶体增殖物激活受体-γ（PPAR-γ）激动剂 可以阻断这种免疫激活，并已被证明可以减弱某些炎症反应， 小胶质细胞和减少浦肯野细胞损失的啮齿动物。这表明，小胶质细胞可能是一个治疗靶点， FASD。已知小胶质细胞塑造小脑中的神经元回路发育和连接， 与小胶质细胞结构动力学有关。乙醇如何影响这些动力学以及它如何影响小胶质细胞- 浦肯野细胞的相互作用是未知的。此外，尚不清楚这些变化何时发生以及如何发生。 在成年期保持或逐渐改变。阐明乙醇如何诱导小胶质细胞的变化 介导小脑浦肯野细胞的一些病理变化可能是理解发病的关键 FASD的病理学。此外，在乙醇暴露过程中，通过调节小胶质细胞的存活和活性， PPAR-γ激动剂可能为这些疾病提供一些答案和潜在的治疗方法。因此，我假设， 乙醇诱导小脑小胶质细胞的神经免疫变化，改变其动力学和与 浦肯野细胞，并通过PPAR-γ激动剂减少小胶质细胞介导的炎症， 酒精的病理作用。为了验证这一假设，我将使用FASD小鼠模型实现两个目标。 第一个将研究发育中的乙醇和PPAR-γ激动剂如何影响小胶质细胞表型， 使用小胶质细胞动力学的体内双光子成像、免疫组织化学和定量真实的时间 PCR法第二个将确定浦肯野细胞和小胶质细胞的相互作用是否在整个生命过程中受到影响， 发育期乙醇暴露和双光子成像的PPAR-γ激动剂给药， 免疫组织化学和电子显微镜。这些实验将阐明小脑的影响， 小胶质细胞对小脑浦肯野细胞的发育后，乙醇暴露，并评估小胶质细胞作为一个 在FASD小鼠模型中减轻疾病病理学的潜在靶点。