Intracellular mechanisms of cocaine-memory reconsolidation

可卡因记忆重建的细胞内机制

• 批准号: 8513130
• 负责人: Audrey M Wells
• 金额: $ 1.53万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2013-12-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8513130
• 关键词: Amygdaloid structure; Animal Model; Anisomycin; Antisense Oligonucleotides; Behavior; Behavioral; Brain; CREB-binding protein; CREB1 gene; Cocaine; Cocaine Dependence; Cocaine Users; Cognitive; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Cyclin-Dependent Kinase 5; Cyclin-Dependent Kinases; Data; Development; Dose; Economics; Environment; Enzymes; Extinction (Psychology); Extracellular Signal Regulated Kinases; Figs - dietary; In Vitro; Infusion procedures; Laboratories; MAP2K1 gene; MEKs; Mediating; Memory; Mitogens; Molecular; Motivation; National Research Service Awards; Pharmaceutical Preparations; Phosphotransferases; Play; Post-Translational Protein Processing; Procedures; Process; Protein Biosynthesis; Public Health; RNA Interference; Rattus; Rehabilitation therapy; Relapse; Research; Response Elements; Retrieval; Rodent Model; Role; Signal Transduction; Signaling Molecule; Small Interfering RNA; Stimulus; Techniques; Testing; Therapeutic; Time; Training; U-0126; United States; Western Blotting; Zinc Fingers; addiction; base; butyrolactone; cocaine exposure; design; drug abstinence; drug addict; drug craving; drug relapse; elk-1 protein; follow-up; human CREBBP protein; indexing; inhibitor/antagonist; insight; kinase inhibitor; long term memory; neurobiological mechanism; neuromechanism; novel; pre-doctoral; prevent; research study; response; social; sound; therapy design; therapy development; transcription factor

DESCRIPTION (provided by applicant): Environmentally-induced drug relapse is a major obstacle in the rehabilitation of cocaine users. Cocaine- associated environments maintain stimulus control over cocaine-induced behavior via context-response- cocaine associations that must be maintained in long-term memory. Thus, treatments that selectively weaken or eliminate such cocaine memories may reduce the propensity to relapse in former drug addicts. Remarkably, cocaine-related long-term memories become labile following retrieval (i.e., memory reactivation), and these memories need to be re-stabilized, or reconsolidated, into long-term storage in order to be maintained. Furthermore, evidence suggests that the basolateral amygdala (BLA) is required for the reconsolidation of cocaine memories that promote drug context-induced cocaine seeking (non-reinforced active lever presses, which provided an index of motivation for cocaine; Fuchs et al., 2009). The proposed NRSA research plan will extend this line of inquiry by exploring the intracellular mechanisms of cocaine-memory reconsolidation in the BLA with a focus on two signaling molecules: extracellular signal-related kinase (ERK) and cyclin-dependent protein kinase 5 (Cdk5). Specific aim 1 will be to test the hypothesis that ERK-mediated signaling in the BLA facilitates the reconsolidation of cocaine memories that are critical for contextual control over instrumental cocaine seeking and will explore possible downstream mechanisms of this phenomenon. Preliminary data show that intra-BLA U0126 (MEK/ERK inhibitor) treatment impairs cocaine-memory reconsolidation in a memory reactivation-dependent manner and disrupts subsequent drug context-induced cocaine seeking (Exp. 1a, in progress). These data provide sound rationale for Exp. 1b, which will utilize quantitative Western blotting to characterize changes in the activation of Elk-1, CREB, and zif268, the transcription factors that are likely activated by ERK in the BLA during memory reconsolidation (Aim 1b). As follow-up, antisense oligonucleotide or small interfering RNA manipulation will be used to assess whether the activation of Elk-1, CREB, or zif268 is required for cocaine-memory reconsolidation and subsequent contextual control over cocaine seeking. Specific aim 2 will be to test the hypothesis that Cdk5 in the BLA interferes with the reconsolidation of cocaine memories that are critical for contextual control over instrumental cocaine seeking. Based on evidence that Cdk5 negatively regulates ERK in vitro, Exp. 2 will assess whether intra-BLA ¿-butyrolactone (Cdk5 inhibitor) treatment will enhance cocaine-memory reconsolidation in a memory reactivation-dependent manner and increase subsequent drug context-induced cocaine seeking. In summary, a combination of sophisticated behavioral procedures and cutting edge molecular techniques will be used to explore the intracellular mechanisms of cocaine-memory reconsolidation, a cognitive process that maintains maladaptive addictive memories and thus stimulates environmentally-induced drug craving and relapse. This research endeavor has the potential to provide novel insights for addiction treatment development.

描述（由适用提供）：环境引起的药物继电器是可卡因使用者康复的主要障碍。可卡因相关的环境通过上下文响应可卡因的关联维持对可卡因诱导的行为的刺激控制，这必须保持在长期记忆中。这是选择性弱或消除这种可卡因记忆的治疗方法可能会降低以前吸毒者的可靠性。值得注意的是，与可卡因相关的长期记忆在检索后（即记忆重新激活）变得不稳定，并且需要将这些记忆重新稳定或重新固定为长期存储，以维持。此外，有证据表明，可卡因记忆的重新固定需要促进药物上下文引起的可卡因寻求的可卡因记忆（非增强的主动杆按压，这为可卡因提供了动机索引； Fuchs等，2009）。拟议的NRSA研究计划将通过探索BLA中可卡因 - 记忆重新溶解的细胞内机制来扩展这种询问，重点是两个信号分子：细胞外信号相关激酶（ERK）和细胞周期蛋白依赖性蛋白蛋白激酶5（CDK5）。具体目的1是测试以下假设：BLA中ERK介导的信号传导促进可卡因记忆的重新整合，这对于对工具可卡因寻求的上下文控制至关重要，并将探索这种现象的可能下游机制。初步数据表明，BLA U0126（MEK/ERK抑制剂）治疗会损害可卡因 - 记忆重新固化的记忆重新激活依赖性方式，并破坏随后的药物上下文引起的可卡因寻求（经验）（经验上的1a，进展中）。这些数据为EXP提供了声音原理。 1b将利用定量的蛋白质印迹来表征ELK-1，CREB和ZIF268激活的变化，ERK在记忆重新保留过程中可能被ERK激活的转录因子可能会激活（AIM 1B）。随后，将使用反义寡核苷酸或小型干扰RNA操纵来评估可卡因 - 记忆重新保留和随后对可卡因寻求可卡因的上下文控制的ELK-1，CREB或ZIF268的激活。具体目的2将测试以下假设：BLA中的CDK5会干扰可卡因记忆的重新整合，这对于对工具可卡因寻求的上下文控制至关重要。基于证据表明CDK5在体外对ERK进行负调节，Exp。 2将评估-bla�-丁乙烯酮（CDK5抑制剂）的治疗是否会以记忆重新激活依赖性方式增强可卡因 - 记忆重新溶解，并增加随后的药物上下文诱导的可卡因寻求。总而言之，将使用复杂的行为程序和尖端分子技术的结合来探索可卡因 - 内存重新溶解的细胞内机制，这是一种认知过程，该过程可维持疾病适应性的附加性记忆，从而刺激环境诱导的药物狂欢和救援。这项研究努力有可能为成瘾治疗发展提供新的见解。