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

描述(由申请人提供)：吸毒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。这项提议的目标是确定蛋白激酶M Zeta(PKMz)，一种与维持海马体长期记忆有关的分子，是否有助于维持在药物使用期间获得的记忆。我们的初步数据表明，PKMz存在于杏仁核中，并为联想、线索条件反射任务维持长期记忆。目前尚不清楚管理基于恐惧的联想任务的相同机制是否对基于奖励的条件性位置偏好任务也很重要。在这个范例中，一只大鼠学会了将阳性增强剂，在这种情况下是可卡因与迷宫的两个手臂之一联系起来。这两只手臂通过使用不同的触觉、嗅觉和视觉线索而彼此不同。测试是衡量每只大鼠在以前与有回报的可卡因相关的手臂上花费的时间。为了研究PKMz在长期记忆中可能起的作用，在每次可卡因手臂训练试验后22小时，将先前被证明是PKMz特异性抑制物的伪底物抑制肽(ZIP)注入双侧杏仁外侧基底核。学位偏好测试将在培训结束后的一天进行。同时，还将进行几个对照实验，以确定Zip是否影响记忆或其他一些行为成分，如主要奖励的体验。除行为外，还将通过Northern、Western、免疫组织化学和原位杂交实验确定CPP任务后不同时间点的蛋白质和mRNA表达水平。了解维持病理性药物使用的机制对于扰乱成瘾循环至关重要。可卡因尤其在美国被广泛使用，与其他滥用物质不同，它缺乏任何药理干预。本工作的目标是揭示因成瘾而改变的关键生化途径，并构建可能的临床干预措施。17B。可卡因是一种被广泛滥用的药物，目前仅在美国就有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，目前还没有有效的药物治疗可卡因依赖。本提案的目标是试图查明导致对可卡因上瘾的潜在生物学机制。我们认为，一种名为蛋白激酶m Zeta的特定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，最终导致成瘾循环。

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.