Molecular genetic analysis of drug addiction

成功治疗吸毒成瘾的一个关键障碍是复发，这往往是由 暴露在压力之下。在动物研究中，通过恢复对药物的反应来模拟复发 自我给药或条件性位置偏爱(CPP)，由药物、线索或压力引起。我们和其他人，有 证明了各种滥用药物，如吗啡、可卡因和最近的尼古丁，都可以 激活转录因子CREB(cAMP反应元件结合蛋白)，这种激活是 对这些药物的后续行为反应是必要的。最近，我们已经证明了CREB是 应激诱导的可卡因条件性位置偏爱(CPP)的恢复所需，但不是药物- 诱导复职。小鼠是一种易于驯服的动物模型，可以用来研究复发 行为水平、神经回路和分子机制。在这个提案中，我们将使用MICE 在CREB(CREB1^突变小鼠)中进行靶向突变的纯合以检验CREB是一种 应激诱导的找药行为恢复所需的中枢信号分子。我们 我将在三个具体目标上检验这一中心假设。在目标1中，我们将调查是否激活了 转录因子CREB在以下各种恢复行为的表现中起关键作用 应激源使用条件性位置偏好范式(CPP)，并确定皮质酮的作用 和CRF在这一反应中。由于应激和可卡因都能恢复野生型小鼠的CPP，这些回路必须 汇聚在最后一条共同的小路上。在目标2中，我们将确定压力与药物之间的特定途径。 诱导恢复，以及汇聚点。此外，逆行追踪研究将使我们能够 以确定这些神经元在恢复电路中关键脑区的准确位置。最后，在 目的3我们将确定这些关键脑区中的哪些基因是CREB的直接靶基因。这些研究 将使我们能够解决药物复发的分子机制。这一信息将是 对开发以调节CREB及其受体为重点的药物治疗方法有价值 靶基因。这些研究可能会产生新的潜在疗法，最终可以转化为 预防人类复发的诊所。

A key obstacle to successful treatment of drug addiction is relapse, which is frequently precipitated by exposure to stress. Relapse has been modeled in animal studies through reinstatement of responses to drug self-administration or conditioned place preference (CPP) by drug, cue or stress. We and others, have demonstrated that a variety of drugs of abuse, such as morphine, cocaine and more recently nicotine, can activate the transcription factor CREB (cAMP response element binding protein) and this activation is necessary for subsequent behavioral responses to these drugs. More recently, we have shown that CREB is required for stress-induced reinstatement of cocaine conditioned place preference (CPP), but not drug- induced reinstatement. The mouse is a tractable animal model that allows for investigation of relapse at the levels of behavior, neural circuitry and molecular mechanisms. In this proposal we will utilize mice homozygous for a targeted mutation in CREB (CREB1^ mutant mice) to test the hypothesis that CREB is a central signaling molecule required for stress inducedreinstatement of drug seeking behavior.We will test this central hypothesis in three specific aims. In Aim 1, we will investigate if activation of the transcription factor CREB is critical for the manifestation of reinstatement behavior following various stressors using a conditioned place preference paradigm (CPP), and determine the role of corticosterone and CRF in this response. As both stress and cocaine reinstate CPP in wild type mice, these circuits must converge on a final common pathway. In Aim 2, we will identify the pathways specific for stress versus drug induced reinstatement, as well as points of convergence. Moreover, retrograde tracing studies will enable us to identify the precise location of these neurons in key brain areas in the reinstatement circuitry. Lastly, in Aim 3 we will determine which genes in these key brain areas are direct CREB target genes. These studies will allow us to address the molecular mechanisms underlying drug relapse. This information will be valuable for the development of pharmacotherapeutic approaches that focus on regulation of CREB and its target genes. These studies may give rise to novel potential therapeutics that can ultimately be translated to the clinic for relapse prevention in humans.