FETAL ALCOHOL SYNDROME--THIRD TRIMESTER MODEL

此续期申请的目的是继续设计的研究 为了提供与酒精背后的机制相关的信息- 相关出生缺陷(ARBD)。这项提议是基于这样的假设 考虑到已经发现的ARBDS的广泛范围，以及 各种条件下可以生产它们，它是非常 造成所有损害的不太可能是一个单一的机制。基于 从上一个供资期间收集的数据，拟议的研究 关注单一的、可靠地产生的数量缺陷，小脑 浦肯野细胞死亡，作为酒精暴露的结果 相当于第三个三个月。这项提案是围绕两个方面设计的 明确的目标。特定目标1有三个目标：1)描述 血液酒精浓度(BAC)依赖浦肯野的时间进程 细胞死亡；2)确定细胞死亡的主要方式 在短暂的酒精暴露之后；以及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，但在较高的BAC中 细胞死亡的主要方式将从凋亡过渡到坏死。 特异性AIM 2将研究浦肯野细胞的机制 通过检验以下四个假说来证明死亡。假设1：酒精 暴露会扰乱关键的凋亡调节蛋白的临界比例， 与浦肯野细胞死亡有关的基因。假设2：饮酒减少 与浦肯野细胞存活相关的神经营养因子。假设 3：替代酒精减少的神经营养因子可以挽救 酒精诱导死亡的浦肯野细胞的一部分。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反应 调整细胞凋亡调节蛋白的平衡。这项建议 整合了活体模型系统(人工饲养新生儿 评估小脑损伤的小鼠)和体外补充 系统(器官型小脑外植体)。各种技术将会 将在建议书中使用，包括光学和电子显微镜，3-D 体视学，流式细胞仪细胞分类和计数，ELISAs，Western 印迹和免疫细胞化学。拟议的研究将产生新的 与酒精如何杀死有丝分裂后神经元相关的数据 发展。对酒精诱导细胞死亡调控的认识 对于制定减少有害影响的策略是必要的 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.