Elucidating the role of the endogenous opioid dynorphin in reward seeking

阐明内源性阿片类强啡肽在寻求奖励中的作用

• 批准号: 10722734
• 负责人: Raajaram Gowrishankar
• 金额: $ 17.46万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10722734
• 关键词: Address; Affect; Amygdaloid structure; Award; Behavior; Behavior Control; Behavioral Model; Biosensor; Brain; Brain region; Budgets; Cessation of life; Committee Members; Consumption; Corpus striatum structure; Cues; Data; Development; Drug usage; Dynorphins; Fentanyl; Fiber; Genetic; Goals; Head; Image; Intake; Internal Ribosome Entry Site; Intervention; Kinetics; Light; Link; Measures; Mediating; Mentors; Messenger RNA; National Institute of Drug Abuse; Neurons; Neurosciences; Operant Conditioning; Opioid; Opioid Antagonist; Oral; Pattern; Persons; Pharmaceutical Preparations; Pharmacology; Phase; Photometry; Population; Postdoctoral Fellow; Predisposition; Principal Investigator; Property; Receptor Signaling; Recreation; Regulation; Relapse; Research; Resolution; Rewards; Role; Scientific Societies; Self Administration; Shapes; Signal Transduction; Substance Use Disorder; Sucrose; Testing; Therapeutic Agents; Training; Validation; calmodulin-dependent protein kinase II; career; career development; conditioning; drug reward; endogenous opioids; fentanyl seeking; fentanyl self-administration; imaging approach; in vivo; in vivo calcium imaging; in vivo two-photon imaging; kappa opioid receptors; knowledge base; neural circuit; neuromechanism; novel; opioid use; opioid use disorder; optogenetics; overdose death; preference; prevent; programs; receptor expression; substance use; symposium; transmission process; two-photon

Project Summary: The primary goal of this training proposal is to understand how dynorphinergic regulation of circuits in the dorsomedial striatum (DMS) enhances seeking behaviors for natural (sucrose) and drug (fentanyl) rewards. Enhanced dynorphin-kappa opioid receptor (dyn-KOR) signaling and aberrant striatal activity is associated with the transition from recreational to persistent opioid-seeking. Yet how dyn neuron activity and subsequent -KOR modulation of the striatum regulates sucrose or fentanyl-seeking is unknown. During the first phase of my postdoctoral research, using a combination of pharmacology, genetics, in vivo photometry and optogenetics, I identified that retrograde dyn transmission at BLA inputs to the DMS enhances BLA-DMS activity, and promotes natural reward-seeking behaviors. However, when exactly dyn neuron activity and release are engaged in reward-seeking is unknown. Hence, I will obtain training in, and use in vivo two-photon imaging to understand how DMS dyn ensembles encode reward-seeking for sucrose, (Aim 1A, K99). I will also extensively characterize and use a novel dyn biosensor using in vivo photometry to determine if the pattern of ensemble activity is reflected in subsequent DMS dyn release, (Aim 1B, K99). Furthermore, because aberrant dyn-KOR signaling is linked to maladaptive opioid-seeking, and my preliminary data suggests a role for dyn-KOR activity to enhance sucrose-seeking, I will develop and use an oral fentanyl self-administration paradigm in conjunction with in vivo photometry to dissect when BLA-DMS terminals are engaged during fentanyl-seeking (Aim 2A, K99). I will also multiplex conditional deletions of dyn in the DMS, or KOR in the BLA, and stimulate dyn release in the DMS, with in vivo photometry during fentanyl-seeking to determine whether enhanced dyn-KOR signaling negatively modulates fentanyl-seeking (Aim 2B, K99). For the R00 “independent” phase of my proposal, I propose to extend the findings from Aims 1 and 2 in my own lab. I will delineate how DMS dyn ensemble activity encodes fentanyl- seeking, manipulate specific ensembles to control behavior via spatial light modulation, and dissect the necessity for BLA inputs in the DMS to induce dyn release during fentanyl-seeking (Aim 3, R00). The proposed studies specifically address how elevated endogenous dynorphin-KOR signaling, via its control of activity in the DMS, regulates natural and fentanyl reward-seeking. During the proposed K99 training period, I will be trained in in vivo two-photon calcium imaging approaches, opioid biosensor imaging and oral self-administration. Additionally, I will actively participate in scientific society conferences, obtain career development training (budgeting and administrative tasks), and continue to further my scholarly knowledge base (planned interactions with my mentor and committee members). Altogether, this award will greatly facilitate the development of my own research program, thereby preparing me for an independent neuroscience career as a principal investigator.

项目概要： 本培训计划的主要目标是了解强啡能回路如何调节 背内侧纹状体（DMS）增强寻求自然（蔗糖）和药物（芬太尼）奖励的行为。 增强的强啡肽-κ阿片受体 (dyn-KOR) 信号传导和异常纹状体活动与 从休闲寻求阿片类药物到持续寻求阿片类药物的转变。然而，dyn 神经元活动和随后的 -KOR 是如何变化的 纹状体的调节调节蔗糖或芬太尼寻求尚不清楚。在我的第一阶段 博士后研究，结合药理学、遗传学、体内光度测定和光遗传学，我 发现 BLA 输入到 DMS 的逆行 dyn 传输增强了 BLA-DMS 活性，并促进 自然的寻求奖励行为。然而，当动态神经元活动和释放发生时， 寻求奖励是未知的。因此，我将接受体内双光子成像的培训并使用它来理解 DMS 动态集成如何编码蔗糖的奖励寻求（目标 1A，K99）。我还将广泛描述 并使用一种新型动态生物传感器，利用体内光度测定来确定整体活动的模式是否是 反映在随后的 DMS dyn 版本中（Aim 1B、K99）。此外，由于异常的 dyn-KOR 信号传导 与适应不良的阿片类药物寻求有关，我的初步数据表明 dyn-KOR 活性可以增强 寻求蔗糖，我将开发并使用口服芬太尼自我给药范例与体内结合 光度测定来剖析 BLA-DMS 终端在芬太尼搜寻过程中何时起作用（目标 2A，K99）。我也会 对 DMS 中的 dyn 或 BLA 中的 KOR 进行多重条件删除，并刺激 DMS 中的 dyn 释放， 在芬太尼寻求过程中使用体内光度测定来确定增强的 dyn-KOR 信号是否会产生负面影响 调节芬太尼寻求（目标 2B，K99）。对于我提案的 R00“独立”阶段，我建议延长 我自己实验室的目标 1 和目标 2 的发现。我将描述 DMS dyn 整体活动如何编码芬太尼 - 寻找、操纵特定的整体以通过空间光调制来控制行为，并剖析其必要性 DMS 中的 BLA 输入可在寻找芬太尼期间诱导 dyn 释放（目标 3，R00）。拟议的研究 具体解决了如何通过控制内源性强啡肽-KOR信号传导的活性来提高内源性强啡肽-KOR信号传导 DMS，调节自然和芬太尼奖励寻求。在拟定的K99培训期间，我将接受培训 体内双光子钙成像方法、阿片类生物传感器成像和口服自我给药。 此外，我将积极参加科学学会会议，获得职业发展培训 （预算和行政任务），并继续深化我的学术知识基础（计划的互动 与我的导师和委员会成员）。总而言之，这个奖项将极大地促进我的发展 自己的研究计划，从而为我作为首席研究员的独立神经科学职业做好准备。