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.

此更新申请的目的是继续研究设计 提供与酒精的潜在机制有关的信息， 相关出生缺陷（ARBD）。 该建议是基于这样的假设， 考虑到已经确定的ARBDS的广泛范围， 各种条件下，他们可以生产，这是非常 不太可能是一个单一的机制造成了所有的损害。 基于 从上一个资助期收集的数据， 专注于一个单一的，可靠的定量缺陷，小脑 浦肯野细胞死亡，由于酒精暴露在部分 相当于第三个三个月。 该提案围绕两个 具体目标。 具体目标1有三个目标：1）表征 血液酒精浓度（BAC）依赖性浦肯野氏 细胞死亡; 2）确定发生的细胞死亡的主要模式 在短暂的酒精暴露之后;以及3）测试 假设中度BAC将诱导浦肯野细胞死亡， 大多数细胞通过凋亡死亡，但在较高的BAC下， 细胞死亡的主要方式将从凋亡转变为坏死。 特异性AIM 2将研究浦肯野细胞的机制 通过测试以下四个假设来确定死亡。 假设1：酒精 暴露破坏了关键凋亡调节蛋白的临界比率， 与浦肯野细胞死亡有关 假设2：酒精降低 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一个 部分浦肯野细胞由酒精诱导的死亡。假设四： 替代神经营养因子将抑制细胞死亡级联反应， 重新调整凋亡调节蛋白的平衡。 该提案 结合了体内模型系统（新生儿的人工饲养 评价小脑损伤）和补充的体外 系统（器官型小脑外植体）。 各种技术将 在提案中使用，包括光学和电子显微镜，三维 体视学、流式细胞术细胞分选和计数、ELISA、Western blot 印迹和免疫细胞化学。 这些研究将产生新的 关于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 与胎儿酒精综合征有关