COCAINE REGULATION OF FOSB SPLICING

描述(改编自申请人的摘要)： 尽管最近的研究揭示了许多关于人类免疫缺陷的分子基础 在吸毒成瘾方面，我们对变化的认识仍然存在许多差距 与慢性药物摄入相关的基因表达。一个突出的问题 在慢性可卡因暴露时发生的改变是在 转录因子DeltaFosB的非常稳定的亚型纹状体，即 通过FosB转录本的选择性剪接产生。DeltaFosB亚型 可能会调节某些神经和行为的改变 吸毒成瘾。在选择性剪接中起作用的因素 尚未鉴定出deltaFasB。要开始理解 导致稳定的DeltaFosB产生的转录后事件，我们 将检验顺式和反式作用因子都很重要的假设 在这项规定中。为了验证这一假设，FosB的突变分析将是 用于鉴定前信使核糖核酸剪接所必需的序列。交联剂 将进行RNA结合研究，以确定哪些反式作用 在从大脑中制备的剪接提取物中，因子与RNA结合。因素 这些研究中确定的基因将在剪接分析中进行测试，以确定 他们对体外拼接进行了监管。这些研究将确定这些因素 在长期使用可卡因的过程中调节剪接所需的，并导致 对所涉及的机制的理解。因为DeltaFosB也被诱导 通过长期服用安非他明等其他滥用药物， 尼古丁和阿片类药物，这项研究的结果也可能导致 更好地理解吸毒成瘾。

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.