Norepinephrine and dopamine dynamics during conditioned place preference and aversion in dorsal hippocampus

背侧海马条件性位置偏好和厌恶过程中去甲肾上腺素和多巴胺的动态变化

• 批准号: 10462416
• 负责人: Avi Kefir Matarasso
• 金额: $ 4.63万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2024-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10462416
• 关键词: Address; Adult; Affect; Agonist; American; Anatomy; Attenuated; Aversive Stimulus; Behavior; Clonidine; Clustered Regularly Interspaced Short Palindromic Repeats; Color; Computer Analysis; Cues; Data; Development Plans; Disease; Dopamine; Dopamine-beta-monooxygenase; Dorsal; Drug usage; Environmental Risk Factor; Enzymes; Exposure to; Fiber; Fluorescence; Genes; Goals; Hippocampus (Brain); Image; In Vitro; Individual; Learning; Measures; Mediating; Mediation; Memory; Mentors; Morphine; Mus; Naloxone; Neurobiology; Neurons; Neuropharmacology; Norepinephrine; Opiate Addiction; Opioid; Optics; Overdose; Pathologic; Persons; Pharmaceutical Preparations; Pharmacology; Photometry; Physiological; Positioning Attribute; Process; Production; Property; Quinpirole; Raclopride; Relapse; Reporting; Research Personnel; Resolution; Rewards; Series; Signal Transduction; Site; Source; Stress; Substance Use Disorder; Synaptic plasticity; Testing; Training; Ventral Tegmental Area; Viral; alpha 2 agonist; antagonist; behavioral pharmacology; biological adaptation to stress; career development; conditioned place preference; conditioning; craving; designer receptors exclusively activated by designer drugs; dopaminergic neuron; drug craving; ex vivo imaging; experimental study; extracellular; in vivo; inhibitor; insight; knock-down; locus ceruleus structure; long term memory; memory acquisition; memory process; monoamine; neuronal cell body; neuroregulation; neurotransmission; noradrenergic; novel; opioid abuse; opioid overdose; opioid use; opioid use disorder; optical imaging; optogenetics; preference; reboxetine; receptor; relating to nervous system; response; reward processing; sensor; spatial memory; spatiotemporal; stress reactivity; two-photon

SUMMARY: In 2017, 19.7 million American adults struggled with substance use disorders, and in 2020, nearly 70,000 people died from opioid-involved overdoses. In opioid dependent individuals, environmental stress perpetuates cravings, and relapse is associated with strong stress responses and reactivity to drug cues. Unexpected reward and aversive stimuli affect ventral tegmental area (VTA) dopamine (DA) neuron activity, and studies neuropharmacological studies have revealed DA released from VTA in dorsal hippocampus (CA1) regulating contextual learning. Locus coeruleus noradrenergic (LC-NE) neurons may also release DA, which has the potential to affect contextual memory and plasticity in CA1. To address how aversive and rewarding contexts affect the opioid seeking, we must understand the neuropharmacological properties of norepinephrine (NE) and DA in opioid-associated contextual memory. The initial process of forming memories is acquisition, while after memories are reactivated, they are considered versatile before being reconsolidated for long term memory. While we know noradrenergic and dopaminergic receptors have been implicated in opioid contextual processing, yet there have not yet been clear pharmacological approaches examining endogenous monoamine release during contextual behavior. The central hypothesis of this proposal is that DA in CA1 is primarily released from the VTA, and only VTA-DA is necessary for, and LC-NE only enhances, memory acquisition and reconsolidation. Aim 1A uses two-photon ex vivo imaging and optogenetics with monoamine sensors to determine spatiotemporal properties of DA release from VTA terminals in dCA1, while Aim 1B will determine spatiotemporal properties of NE and DA release from LC in CA1. Using ex vivo pharmacology, we will elucidate which dopaminergic and noradrenergic mechanisms in CA1 influence the release of DA and NE from their source regions. Aim 2 will tests the necessity of NE and DA sources for morphine or naloxone place preference reconsolidation and the necessity of NE and DA for preference acquisition. In Aim 2A, we will simultaneously measure NE and DA dynamics as we pharmacologically inhibit their soma using chemogenetics to determine the necessity of VTA and LC in memory reconsolidation of a conditioned place preference or aversion. In Aim 2B, we will test the necessity of NE and DA for memory acquisition of conditioned preference or aversion by using CRISPR gene-editing to knockdown Dbh (NE-producing enzyme) in LC and Th (DA-producing enzyme) in VTA. This series of in vitro and in vivo neuropharmacology experiments will elucidate monoamine sufficiency and necessity in contextual processing to further our understanding of the influence of context in opioid seeking. This proposal will serve as training in neuropharmacology, two-photon optical imaging, optogenetics, chemogenetics, gene-editing, and behavioral pharmacology approaches. Ultimately, the training outlined in this proposal will enhance my career development plan as I pursue a position as an independent, academic researcher.

摘要：2017年，1970万美国成年人与物质使用障碍作斗争，2020年， 7万人死于阿片类药物过量。在阿片类药物依赖者中， 持续的渴望，复发与强烈的压力反应和对药物线索的反应有关。 意外的奖赏和厌恶刺激影响腹侧被盖区（VTA）多巴胺（DA）神经元的活动， 研究神经药理学研究显示DA从背侧海马（CA 1）的VTA释放 调节情境学习。蓝斑去甲肾上腺素能（LC-NE）神经元也可能释放DA， 可能影响CA 1的背景记忆和可塑性。为了说明令人厌恶和有益的环境 影响阿片寻求，我们必须了解去甲肾上腺素（NE）的神经药理学特性， 阿片相关背景记忆中的DA记忆形成的最初过程是获得， 当记忆被重新激活时，它们在被重新巩固为长期记忆之前被认为是多功能的。 虽然我们知道去甲肾上腺素能和多巴胺能受体与阿片类物质的背景处理有关， 然而，还没有明确的药理学方法来检查内源性单胺释放 在上下文行为中。这一建议的中心假设是，CA 1中的DA主要从 VTA，并且只有VTA-DA是必需的，并且LC-NE仅增强记忆获取和再巩固。 Aim 1A使用双光子离体成像和具有单胺传感器的光遗传学来确定 dCA 1中VTA终末DA释放的时空特性，而Aim 1B将决定时空 CA 1中LC释放NE和DA的性质。使用离体药理学，我们将阐明 CA 1的多巴胺能和去甲肾上腺素能机制影响DA和NE从其来源的释放 地区目的2：探讨吗啡或纳洛酮位置偏爱的NE和DA来源的必要性 重新整合和必要性的NE和DA的优先收购。在目标2A中，我们将同时 当我们使用化学遗传学抑制NE和DA的索马时， VTA和LC在条件性位置偏爱或位置厌恶记忆再巩固中的必要性。在Aim中 2B，我们将测试NE和DA对于条件性偏好或厌恶的记忆获得的必要性， 使用CRISPR基因编辑来敲低LC中的Dbh（产生NE的酶）和Th（产生DA的酶） 在VTA。这一系列的体外和体内神经药理学实验将阐明单胺的充足性 以及语境加工的必要性，以进一步理解语境对阿片类药物寻求的影响。 该提案将作为神经药理学，双光子光学成像，光遗传学， 化学遗传学、基因编辑和行为药理学方法。最后，本文中概述的培训 我的建议将加强我的职业发展计划，因为我追求的立场，作为一个独立的，学术 研究员