FETAL ALCOHOL SYNDROME--THIRD TRIMESTER MODEL

胎儿酒精综合症——妊娠晚期模型

• 批准号: 6509116
• 负责人: JAMES R WEST
• 金额: $ 24.57万
• 依托单位: TEXAS A&M UNIVERSITY HEALTH SCIENCE CTR
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-09-01 至 2004-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6509116
• 关键词: alcoholic beverage consumption; apoptosis; cell death; cerebellar Purkinje cell; cytotoxicity; developmental neurobiology; dosage; electron microscopy; enzyme linked immunosorbent assay; fetal alcohol syndrome; flow cytometry; gestational age; immunocytochemistry; infant animal; laboratory rat; light microscopy; necrosis; neurons; radioimmunoassay; statistics /biometry; western blottings

The purpose of this renewal application is to continue studies designed to provide information related to mechanisms that underlie alcohol- related birth defects (ARBDs). The proposal is based on the supposition that given the wide spectrum of ARBDS that have been identified, and the variety of conditions under which they can be produced, it is extremely unlikely that a single mechanism underlies all of the damage. Based on data gathered from the preceding funding period, the proposed studies focus on a single, reliably produced quantitative deficit, cerebellar Purkinje cell death, as a consequence of alcohol exposure during part of the third trimester equivalent. The proposal is designed around two Specific Aims. SPECIFIC AIM 1 has three objectives: 1) to characterize the time course for blood alcohol concentration (BAC)-dependent Purkinje cell death; 2) to identify the primary mode of cell death that occurs following a short episode of alcohol exposure; and 3) test the hypothesis that moderate BACs will induce Purkinje cell death with the majority of cells dying via apoptosis, but at higher BACs the predominant mode of cell death will sift from apoptosis to necrosis. SPECIFIC AIM 2 will investigate mechanisms underlying Purkinje cell death by testing the following four hypotheses. Hypothesis 1: Alcohol exposure disrupts critical ratios of key apoptotic regulator proteins, that are involved in Purkinje cell death. Hypothesis 2: Alcohol reduces neurotrophic factors associated with Purkinje cell survival. Hypothesis 3: Replacing neurotrophic factors reduced by alcohol can rescue a portion of Purkinje cells from alcohol-induced death. Hypothesis 4: Replacing neurotrophic factors will suppress the cell death cascade by readjusting the balance of apoptotic regulator proteins. The proposal incorporates an in vivo model system (the artificial rearing of neonatal rat pups to evaluate cerebellar damage) and a complementary in vitro system (organotypic cerebellar explants). A variety of techniques will be used in the proposal, including light and electron microscopy, 3-D stereology, cell sorting and counting by flow cytometry, ELISAs, Western blots, and immunocytochemistry. The proposed studies will generate new data related to how alcohol kills postmitotic neurons during development. Understanding the regulation of alcohol-induced cell death is necessary for developing strategies to reduce the deleterious effects associated with fetal alcohol syndrome.

此更新应用的目的是继续设计 提供与酒精基础机制有关的信息 相关的先天缺陷（ARBD）。 该提议基于假设 鉴于已经确定的广泛的ARBD， 它们可以生产的各种条件，这是极其的 单一机构不太可能构成所有损害。 基于 拟议的研究从前一个资金时期收集的数据 专注于单一可靠地产生的定量赤字，小脑 Purkinje细胞死亡，由于酒精暴露在零件期间 第三学期等效。 该提案的设计左右 具体目标。 特定目标1具有三个目标：1）要表征 血液酒精浓度的时间过程（BAC）依赖性Purkinje 细胞死亡； 2）确定发生的主要细胞死亡模式 经过短暂的酒精暴露； 3）测试 假设中等BAC会诱导Purkinje细胞死亡 大多数细胞因凋亡而死，但在较高的BAC下 细胞死亡的主要模式将从细胞凋亡到坏死。 特定的目标2将研究浦肯野细胞的基础机制 通过检验以下四个假设来死亡。 假设1：酒精 暴露会破坏关键凋亡调节蛋白的关键比率， 涉及Purkinje细胞死亡。 假设2：酒精减少 与Purkinje细胞存活相关的神经营养因子。 假设 3：替代酒精降低的神经营养因素可以营救 来自酒精引起的死亡的珀kinje细胞的一部分。假设4： 替换神经营养因素将抑制细胞死亡级联 重新调整凋亡调节蛋白的平衡。 提案 结合了体内模型系统（新生儿的人工饲养 大鼠幼崽评估小脑损伤）和体外互补 系统（器官小脑外植体）。 各种技术将 在提案中使用，包括光和电子显微镜，3-D 立体学，细胞分选和计数流式细胞术，ELISA，西方 印迹和免疫细胞化学。 拟议的研究将产生新的 与酒精如何杀死后有丝分裂神经元有关的数据 发展。 了解酒精诱导的细胞死亡的调节 制定减少有害影响的策略是必要的 与胎儿酒精综合征有关。