COCAINE REGULATION OF FOSB SPLICING

FOSB 剪接的可卡因监管

• 批准号: 6260335
• 负责人: Judith Ann Potashkin
• 金额: $ 7.8万
• 依托单位: ROSALIND FRANKLIN UNIV OF MEDICINE & SCI
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-30 至 2002-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6260335
• 关键词: HeLa cells; RNA splicing; RNase protection assay; cocaine; drug abuse; gel mobility shift assay; laboratory rat; messenger RNA; neuropharmacology; nucleus accumbens; protein isoforms; transcription factor; transfection

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.

描述（改编自申请人的摘要）： 尽管最近的研究揭示了大量关于分子基础的信息 吸毒成瘾，我们对吸毒成瘾的变化的理解仍然存在许多差距 与慢性药物摄入相关的基因表达。一位杰出的 长期接触可卡因时发生的改变是在 转录因子 deltaFosB 非常稳定的亚型纹状体 由 fosB 转录物的选择性剪接产生。 deltaFosB 同工型 可能介导一些神经和行为改变 吸毒成瘾。影响选择性剪接的因素 deltaFasB 尚未被鉴定。开始了解 导致稳定 deltaFosB 产生的转录后事件，我们 将检验顺式作用因子和反式作用因子都很重要的假设 在此规定中。为了检验这一假设，将进行 fosB 的突变分析 用于识别前 mRNA 剪接所必需的序列。交联 将进行RNA结合研究以确定哪些反式作用 因子与从大脑中制备的剪接提取物中的 RNA 结合。因素 这些研究中确定的基因将在剪接试验中进行测试，以确定是否 它们在体外调节剪接。这些研究将确定因素 长期使用可卡因期间剪接的调节所需，并导致 了解所涉及的机制。因为 deltaFosB 也会被诱导 长期服用其他滥用药物，例如安非他明， 尼古丁和阿片类药物，这项研究获得的结果也可能导致 更好地了解毒瘾。