Sage in Cerebellum After Alcohol Exposure

描述（由申请人提供）：过量饮酒已知对胎儿、成人和衰老的神经系统有毒性作用。小脑神经元在整个发育过程中受到酒精管理的破坏性影响，这是公认的。由于小脑的主要功能是通过整合感觉输入和运动输出来调节运动和平衡，因此任何成分神经元的损伤都可能导致运动协调障碍。酒精引起的小脑损伤是许多基因表达累积变化的结果，这似乎是合理的。迄今为止，只有一小部分组成人类和小鼠基因组的基因被单独或通过DNA微阵列检测酒精引起的变化。由于DNA微阵列技术是一个“封闭”系统，只能检测已知基因，因此我们建议使用基因表达序列分析（SAGE）对酒精暴露小鼠小脑中的基因表达进行全局分析。SAGE是一个“开放”系统，能够检测和数字化量化已知和未知的基因。拟议的实验包括在产后早期、成年期和衰老期（在大量饮酒后不久）对常规小脑SAGE文库进行构建、测序和生物信息学分析。由此产生的酒精诱导基因表达变化（包括新基因）的全面定量SAGE数据库将通过NCBI网站发布。未来的长期实验，超出本R03的范围，并作为R01计划提交的一部分，将旨在(1)在酒精暴露后的几个时间间隔内构建“定时”小脑SAGE文库；(2)慢性酒精暴露小鼠小脑SAGE文库的制备；(3)充分表征由于酒精摄入而显著改变表达的新型小脑基因；(4)研究旨在逆转酒精摄入对小脑神经元毒性作用的实验策略。

DESCRIPTION (provided by applicant): Excessive consumption of alcohol is known to have toxic effects on the fetal, adult and aging nervous system. It is well established that cerebellar neurons are subject to the damaging effects of alcohol administration throughout development. Because the major function of the cerebellum is to modulate movement and balance by integrating sensory input and motor output, any injury to component neurons is likely to lead to the impairment of motor co-ordination. It is plausible that alcohol-induced cerebellar damage is the result of cumulative changes in the expression of many genes. To date, only a fraction of the genes comprising the human and mouse genomes have been assayed for alcohol-induced changes, either individually or by DNA microarrays. Because DNA microarray technology is a 'closed' system, capable of detecting only known genes, we propose to conduct a global analysis of gene expression in the alcohol-exposed mouse cerebellum using Serial Analysis of Gene Expression (SAGE). SAGE is an 'open' system capable of detecting and digitally quantifying both known and, as yet, unknown genes. The proposed experiments involve the construction, sequencing and bioinformatic analysis of routine cerebellar SAGE libraries during the early postnatal period, adulthood and senescence, shortly after exposure to "binge" amounts of alcohol. The resulting comprehensive and quantitative SAGE database of alcohol-induced changes in gene expression, which includes that of novel genes, will be disseminated via the NCBI website. Future long-term experiments, beyond the scope of this R03 and proposed as part of a planned R01 submission, will be aimed at (1) construction of "timed" cerebellar SAGE libraries at several time intervals after alcohol exposure; (2) preparation of cerebellar SAGE libraries from mice chronically exposed to alcohol; (3) the full characterization of novel cerebellar genes whose expression is substantially altered as a result of alcohol administration, and (4) the examination of experimental strategies aimed at reversing the toxic effects of alcohol ingestion on cerebellar neurons.