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.

项目摘要： 该培训建议的主要目的是了解对电路中电路的调节 背侧纹状体（DMS）增强了寻求天然（蔗糖）和药物（芬太尼）奖励行为的寻求行为。 增强的Dynorphin-kappa Oioid受体（Dyn-Kor）信号传导和异常纹状体活性与 从娱乐性到持续的阿片类药物寻求的过渡。然而，Dyn神经元的活动和随后的活动如何 纹状体调节蔗糖或寻求芬太尼的调节尚不清楚。在我的第一阶段 博士后研究，结合了药理学，遗传学，体内光度法和光遗传学的组合 确定在BLA输入到DMS时逆行DYN传播增强了BLA-DMS活动，并促进 自然的寻求奖励行为。但是，当确切的dyn神经元活动和释放参与时 寻求奖励是未知的。因此，我将获得培训，并使用体内两光子成像来理解 DMS Dyn合奏如何编码寻求蔗糖的奖励（AIM 1A，K99）。我也将广泛地描述 并使用新型的DYN生物传感器使用体内光度法来确定集合活动的模式是否为 反映在随后的DMS Dyn版本中（AIM 1B，K99）。此外，因为异常的dyn-kor信号是 与适应不良的阿片类药物相关，我的初步数据表明了Dyn-Kor活动的作用，以增强 寻求蔗糖，我将开发和使用口服芬太尼自我管理范式与体内结合 在寻求芬太尼期间参与BLA-DMS终端时进行剖析的光度法（AIM 2A，K99）。我也会 dms或bla中的dyn的多重条件缺失，并刺激DMS中的Dyn释放， 寻求芬太尼期间的体内光度法，以确定增强的Dyn-Kor信号是否有负 调节寻求芬太尼（AIM 2B，K99）。对于我的提案的R00“独立”阶段，我提议扩展 AIMS 1和2的发现在我自己的实验室中。我将描述DMS Dynelemble活动如何编码芬太尼 - 寻求，操纵特定的合奏以通过空间光调制来控制行为，并剖析必要的 对于DMS中的BLA输入，可以在寻求芬太尼期间诱导Dyn释放（AIM 3，R00）。提出的研究 特别针对内源性驱动器信号的升高，是如何通过控制其活性的控制 DMS，调节自然和芬太尼寻求奖励。在拟议的K99培训期间，我将接受培训 体内两光子钙成像方法，阿片生物传感器成像和口服自我给药。 此外，我将积极参加科学社会会议，获得职业发展培训 （预算和行政任务），并继续进一步进一步发展我的科学知识库（计划的互动） 与我的导师和委员会成员一起）。总之，这个奖项将极大地支持我的发展 自己的研究计划，从而为我做好了为独立的神经科学事业做好准备。