FETAL ALCOHOL SYNDROME--THIRD TRIMESTER MODEL

此更新申请的目的是继续设计的研究 提供与酒精潜在机制相关的信息 相关出生缺陷（ARBD）。 该提案基于以下假设 鉴于已确定的 ARBDS 范围广泛，并且 它们可以在多种条件下生产，这是极其 不可能有单一机制造成所有损害。 基于 从之前的资助期间收集的数据、拟议的研究 专注于单一的、可靠产生的数量赤字，小脑 浦肯野细胞死亡，由于部分期间接触酒精而导致 妊娠晚期的等效值。 该提案围绕两个方面进行设计 具体目标。 具体目标 1 具有三个目标：1) 表征 血液酒精浓度 (BAC) 依赖性浦肯野病的时间进程 细胞死亡； 2）确定发生的细胞死亡的主要模式 短暂饮酒后； 3）测试 假设适度的 BAC 会诱导浦肯野细胞死亡 大多数细胞通过凋亡而死亡，但在 BAC 较高时 细胞死亡的主要模式将从细胞凋亡转向坏死。 具体目标 2 将研究浦肯野细胞的潜在机制 通过检验以下四个假设来判断死亡。 假设1：酒精 暴露破坏了关键凋亡调节蛋白的关键比例， 参与浦肯野细胞死亡。 假设2：酒精会减少 与浦肯野细胞存活相关的神经营养因子。 假设 3：替代因酒精而减少的神经营养因子可以挽救 部分浦肯野细胞因酒精引起的死亡。假设4： 替代神经营养因子将通过以下方式抑制细胞死亡级联： 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） 幼鼠评估小脑损伤）和体外补充 系统（器官型小脑外植体）。 多种技术将 在提案中使用，包括光学和电子显微镜、3-D 体视学、通过流式细胞术、ELISA、Western 进行细胞分选和计数 印迹和免疫细胞化学。 拟议的研究将产生新的 有关酒精如何杀死有丝分裂后神经元的数据 发展。 了解酒精诱导的细胞死亡的调节 对于制定减少有害影响的战略是必要的 与胎儿酒精综合症有关。

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.