PKM zeta-dependent maintenance of LTM in cocaine conditioned place preference

药物成瘾是当今美国最昂贵和最具破坏性的医疗问题之一。每年有数十万人死于与毒品或酒精有关的疾病。这项提议的目的是确定蛋白激酶M zeta（PKMz），一种与维持海马长期记忆有关的分子，是否有助于维持药物使用期间获得的记忆。我们的初步数据表明，PKMz存在于杏仁核，并保持长期记忆的联想，线索条件反射任务。目前尚不清楚是否同样的机制，管理恐惧为基础的联想任务也是重要的奖励为基础的，条件的位置偏好任务。在这个范例中，一只老鼠把阳性的兴奋剂（在这个例子中是可卡因）和迷宫的两个臂中的一个联系起来。learns.to这两个手臂通过使用不同的触觉、嗅觉和视觉线索而彼此不同。测试是测量每只大鼠在先前与奖励药物可卡因相关的手臂上花费的时间。为了研究PKMz可能对长期记忆的作用，在每次可卡因臂训练试验后22小时，将先前显示为PKMz特异性抑制剂的假底物抑制肽（zip）双侧注入基底外侧杏仁核。地点偏好测试将在培训结束后的某一天进行。与此同时，将进行几个对照实验，以确定zip是否影响记忆或其他行为成分，如主要奖励的体验。除行为外，还将在CPP任务后的不同时间点通过北方、西方、免疫组织化学和原位杂交实验测定蛋白质和mRNA水平的表达。了解维持病理性药物使用的机制对于破坏成瘾循环至关重要。尤其是可卡因在美国被广泛使用，与其他滥用物质不同，可卡因缺乏任何药物干预。目前工作的目标是揭示成瘾改变的关键生化途径，并构建可能的临床干预措施。17 b.可卡因是一种广泛滥用的药物，仅在美国就有230万可卡因使用者，仍然是一个重要的公共卫生问题。不幸的是，没有有效的药物治疗可用于可卡因依赖。本建议的目的是试图确定导致可卡因成瘾的潜在生物学机制。我们相信，一种叫做蛋白激酶m zeta的特殊分子对长期记忆至关重要，它可能通过记忆机制导致渴望、复发并最终形成成瘾循环。

DESCRIPTION (provided by applicant): Drug addiction is one of the most costly and devastating medical problems in the United States today. Hundreds of thousands of people die from drug or alcohol related illnesses each year. The goal of this proposal is to determine if protein kinase M zeta (PKMz), a molecule implicated in maintaining long term memory in the hippocampus, contributes to sustaining memories acquired during drug use. Our preliminary data suggests that PKMz is present in the amygdala and maintains long term memory for an associative, cued conditioning task. It is not known if the same mechanisms governing a fear based associative task are also important for a reward based, conditioned place preference task. In this paradigm a rat learns.to associate a positive reinforcer, in this case cocaine with one of the two arms of a maze. The two arms are distinctive from one another through the use of a different tactile, olfactory, and visual cues. Testing is a measure of the time each rat spends in the arm previously associated with the rewarding drug cocaine. To investigate the role PKMz may have on the-long term memory, a pseudosubstrate inhibitory peptide (zip), previously shown to be a PKMz specific inhibitor, will be bilaterally infused into the basal lateral amygdala twenty-two hours after each cocaine-arm training trial. Testing for place preference will occur one day upon completion of training. In parallel several control experiments will be done to determine if zip is affecting memory or some other behavioral component such as the experience of the primary reward. In addition to behavior, expression of protein and mRNA levels will be determined at different time points following the CPP task by northern, western, immunohistochemistry and in-situ hybridization experiments. Understanding the mechanisms which sustain pathological drug use is critical to disrupting the addictive cycle. Cocaine in particular is widely used in the United States and unlike other substances of abuse, lack any pharmacological intervention. The goal of the present work is to uncover the critical biochemical pathways that are altered by addiction and construct possible clinical interventions. 17b. Cocaine is a widely abused drug and continues to be an important public health problem with 2.3 million current cocaine users in the U.S. alone. Unfortunately, no effective pharmacological treatments are available for cocaine dependence. The goal of the present proposal is to attempt to identify the underlying biological mechanisms contributing to the addiction to cocaine. We believe that a particular molecule called protein kinase m zeta that is critical for long term memory may, through memory mechanisms contribute to craving, relapse and ultimately the cycle of addiction.