Chemical-genetic dissection of the role of lateral habenula in cocaine relapse

描述(由申请人提供)：治疗可卡因成瘾的一个重要问题是即使在戒毒多年后也再次使用可卡因。暴露在压力之下在促使复发方面起着重要作用。因此，研究压力诱发复发背后的神经回路对于开发有效的成瘾治疗方法是必要的。虽然大脑的许多区域参与了应激诱发复发的表达，但对外侧缰核的确切作用知之甚少。利用自我管理-恢复程序，我们建议使用最先进的遗传和化学靶向方法，直接研究这一大脑区域在应激诱导的可卡因复发中的作用。在这个方案中，我们将利用DREADD(由设计师药物独有激活的设计师受体)技术，通过该技术，我们将以一种选择性、快速和可逆的方式操纵外侧缰核神经元，并研究这些操纵对应激诱导的复发的影响。在目前神经科学领域可用的方法下，研究特定突触连接对特定行为的贡献一直是一个挑战。我们建议利用一种独特的双病毒策略，将漂浮的、倒置的DREADD引入外侧缰核神经元，并结合逆行运输的表达Cre重组酶的犬腺病毒2，来研究与可卡因复发有关的神经元回路。在特定的目标1中，我们将研究外侧缰核神经元活动的瞬时调节是否影响应激诱导的可卡因寻找的恢复。在特定的目标2中，我们将验证双病毒方法来解剖突触连接，并研究从外侧缰核到单胺能核的投射神经元是否参与应激诱导的可卡因寻求的恢复。除了提供关于外侧缰核在可卡因复发中的作用的新信息外，我们还将验证双病毒策略的使用，该策略有可能被广泛用作研究的有价值的工具 清醒、行为正常的动物的神经元回路。我们的研究结果将为应激诱发复发的神经回路提供新的见解，并可能为治疗可卡因成瘾的治疗选择开辟新的途径。

DESCRIPTION (provided by applicant): An important problem in the treatment of cocaine addiction is relapse to cocaine use even years after abstinence. Exposure to stress plays an important role in precipitating relapse. Thus, studying the neuronal circuitry that underlies stress-induced relapse is necessary for the development of effective treatments for addiction. While many brain regions contribute to the expression of stress-induced relapse, very little is known about the precise role of the lateral habenula. Using the self-administration-reinstatement procedure, we propose to directly examine the role of this brain region in stress-induced relapse to cocaine seeking using state-of-the-art genetic and chemical targeting approaches. In this proposal, we will utilize the DREADD (Designer Receptors Exclusively Activated by Designer Drugs) technology, by which we will manipulate lateral habenula neurons in a manner that is selective, rapid and reversible and study the effect of these manipulations on stress-induced relapse. With the methods currently available in the field of neuroscience, it has been a challenge to study the contribution of specific synaptic connections involved in a specific behavior. We propose to utilize a unique dual-virus strategy in which we will use a combination of floxed, inverted DREADD's introduced into lateral habenula neurons and a retrogradely transported canine adenovirus 2 engineered to express Cre recombinase, to study neuronal circuits involved in cocaine relapse. In specific aim 1, we will investigate whether transient modulation of lateral habenula neuronal activity influences stress-induced reinstatement of cocaine-seeking. In specific aim 2, we will validate the dual-virus approach to dissect synaptic connections and study whether projection neurons from the lateral habenula to monoaminergic nuclei are involved in stress-induced reinstatement of cocaine seeking. In addition to providing novel information on the role of the lateral habenula in cocaine relapse, we will validate the use of the dual-virus strategy which has the potential to be widely used as a valuable tool in studying neuronal circuits in awake, behaving animals. Results from our studies will offer new insights into the neuronal circuitry that underlies stress-induced relapse and possibly open up new avenues for therapeutic options for the treatment of cocaine addiction.