Studies of the Mouse Kappa Opioid Receptor Gene

描述(申请人提供)：本申请是为续期NIDA赞助的K02(DA13926)独立科学家奖。目前的K02奖(2002-2007年)使PI能够发展她的独立研究生涯，专注于药物受体，特别是阿片受体的表达的遗传编程。已知阿片受体的数量对阿片类化合物的镇痛作用至关重要。这种控制是通过基因编程来指示细胞正确和特异地表达这些基因(转录控制)，并根据需要从mRNAs(转录后)产生这些蛋白质。在以往的研究中，主要任务是剖析负责阿片受体基因转录和转录后调控的基本元件，包括确定转录调控所需的DMA序列和蛋白质因子，识别调控神经元中阿片受体蛋白产生的速率、地点和时间的转录后事件，并从根本上剖析其调控的重要机制。在2002-2005年间，在K02的支持下，国际文学联合会的作品发表在35篇论文中。更新建议将继续系统地研究这些生物过程的机制基础，以及在动物中的整合，以了解阿片受体表达的动态平衡控制。因此，研究项目的总体方向保持不变。具体目标是：1)通过关注转录因子如何与KOR基因的染色质相互作用，导致启动子区的染色质重塑，来扩大对KOR基因转录调控的研究；2)通过重点控制5‘-和3’-非翻译区调控神经元中KOR蛋白的转录后事件，识别调节神经元中KOR蛋白产生的转录后事件；以及3)建立转基因小鼠模型，以回答这些调控事件是否以及如何影响阿片类药物的药理效应和问题的表现，如成瘾和耐受。这些研究对于确定阿片受体表达的遗传编程中的关键步骤具有重要的临床意义，这可能有助于阐明药物滥用问题干预的潜在靶点，并从根本上适用于理解具有重大药理学和神经学后果的神经元隔室特异性蛋白质表达所需的关键生物学过程。这一奖项也对PI作为药物滥用研究领域的独立科学家的持续职业发展至关重要。

DESCRIPTION (provided by applicant): This application is for a renewal of a NIDA sponsored K02 (DA13926) Independent Scientist Award. The current K02 award (2002-2007) allowed the PI to develop her independent research career that focuses on genetic programming for the expression of drug receptors, in particular, the opioid receptors. It is known that the amounts of opioid receptors are crucial for the analgesic effects of opioid compounds. The control is through genetic programming that instructs the cells to express these genes properly and specifically (transcriptional control) and to produce these proteins from mRNAs (post-transcriptionally) according to the needs. In the previous studies, the major task was to dissect the basic elements responsible for transcriptional and post-transcriptional regulation of opioid receptor genes, that include determining DMA sequences and protein factors required for transcriptional control, identification of post-transcriptional events that modulate the rate, the place and the time of opioid receptor protein production in neurons, and dissecting fundamentally important mechanisms in their regulation. Over the period of 2002-2005 supported by this K02, the Pi's works were published in 35 papers. The renewal proposal will continue systemic investigation into the mechanistic basis of these biological processes and the integration in the animals for understanding the homeostatic control of opioid receptor expression. Thus, the overall direction of the research project remains the same. Specific aims are: 1) to extend studies of transcriptional regulation of KOR gene by focusing on how transcription factors interact with chromatin of the KOR gene, leading to chromatin remodeling of the promoter regions; 2) to identify post-transcriptional events that regulate KOR protein production in neurons by focusing on the control of mRNA transport and local translation in neurons regulated by the 5'- and the 3'-untranslated regions; and 3) to produce transgenic mouse models to answer whether and how these regulatory events affect the manifestation of pharmacological effects and problems of opioids, such as addiction and tolerance. These studies are clinically important for the identification of crucial steps in genetic programming of opioid receptor expression that may shed light on potential targets of interventions for drug abuse problems, and are fundamentally applicable for the understanding of critical biological processes required for neuronal compartment-specific expression of proteins that are of significant pharmacological and neurological consequences. This award is also essential for the Pi's continual career development as an independent scientist in the field of drug abuse research.