COCAINE REGULATION OF FOSB SPLICING

描述（改编自申请人摘要）： 虽然最近的研究已经揭示了很多关于 吸毒成瘾，许多差距仍然在我们的理解的变化， 与慢性药物摄入相关的基因表达。一个突出 慢性可卡因暴露后发生的变化是在大脑中的积累 纹状体非常稳定的转录因子deltaFosB亚型， 由fosB转录物的选择性剪接产生。deltaFosB亚型 可能介导一些神经和行为的修改， 毒瘾在选择性剪接中起作用的因素 deltaFasB尚未被鉴定。为了开始理解 转录后事件，导致稳定deltaFosB的生产，我们 将检验顺式和反式作用因子都很重要的假设 在这一规定中。为了验证这一假设，将对fosB进行突变分析。 用于鉴定前体mRNA剪接所必需的序列。交联 和RNA结合研究将进行，以确定哪些反式作用 在从大脑中提取的剪接提取物中，因子与RNA结合。因素 将在剪接试验中检测这些研究中鉴定的基因，以确定 它们调节体外剪接。这些研究将确定 在慢性可卡因使用期间调节剪接所需的，并导致 对相关机制的理解。因为deltaFosB也被诱导 通过长期服用其他滥用药物，如安非他明， 尼古丁和阿片类药物，这项研究中获得的结果也可能导致 更好地了解药物成瘾。

DESCRIPTION(Adapted from applicant's abstract): Although recent studies have revealed a great deal about the molecular basis of drug addiction, numerous gaps remain in our understanding of the changes in gene expression that are associated with chronic drug intake. One prominent alteration that occurs upon chronic cocaine exposure is accumulation in the striatum of very stable isoforms of the transcription factor deltaFosB that is produced by alternative splicing of the fosB transcript. The deltaFosB isoforms may mediate some of the neural and behavioral modivications that occur with drug addiction. The factors that play a role in the alternative splicing of deltaFasB have not yet been identified. To begin to understand the posttranscriptional events that lead to the production of stable deltaFosB, we will test the hypothesis that both cis- and trans-acting factors are important in this regulation. To test this hypothesis, mutation analysis of fosB will be used to identify the sequences essential for pre-mRNA splicing. Cross linking and RNA binding studies will be undertaken to determine which trans acting factors bind to the RNA in splicing extracts prepared from brains. Factors identified in these studies will be tested in splicing assays to determine if they regulate splicing invitro. These studies will identify the factors required for the regulation of splicing during chronic- cocaine use and lead to an understanding of the mechanisms involved. Because deltaFosB is also induced by chronic administration of other drugs of abuse, such as amphetamine, nicotine and opiates, the results obtained in this study may also lead to a better understanding of drug addiction.