In vivo calcium imaging during appetitive learning in HIV Tat transgenic mice exposed to cannabis

暴露于大麻的 HIV Tat 转基因小鼠食欲学习过程中的体内钙成像

• 批准号: 10696442
• 负责人: Sylvia Fitting
• 金额: $ 23.33万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-15 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10696442
• 关键词: 2-arachidonylglycerol; Adult; Age Months; Animal Testing; Animals; Anxiety; Association Learning; Astrocytes; Behavior; Behavior assessment; Behavioral; Brain; Brain region; CNR1 gene; CNR2 gene; Calcium; Cannabinoids; Cannabis; Cells; Chronic; Communication; Conditioned Stimulus; Cues; Desire for food; Discrimination Learning; Disease; Dose; Drug usage; Endocannabinoids; Enzymes; Experimental Designs; Exposure to; Female; GPR55 receptor; Genetic Transcription; Genotype; Goals; HIV; HIV-1; HIV-associated neurocognitive disorder; Head; Human; Immunohistochemistry; Impaired cognition; Inflammation; Inflammatory; Injections; Learning; Ligands; Liquid Chromatography; Memory; Mental Depression; Microglia; Mouse Protein; Mus; Nausea; Nerve Degeneration; Neurocognition; Neuronal Dysfunction; Neurons; Nucleus Accumbens; Outcome; Pathology; Performance; Persons; Pharmaceutical Preparations; Pisum sativum; Play; Prefrontal Cortex; Prevalence; Process; Proteins; Recovery; Reporting; Rewards; Role; Sucrose; Symptoms; Synapses; System; THC exposure; Therapeutic; Training; Trans-Activators; Transcription Coactivator; Transcriptional Regulation; Transgenic Mice; Transgenic Organisms; Viral; Virus Diseases; Visualization; Work; antiretroviral therapy; behavior test; cannabinoid treatment; chronic pain; cognitive function; cognitive performance; cohort; common symptom; endogenous cannabinoid system; excitotoxicity; humanized mouse; in vivo; in vivo calcium imaging; innovation; male; marijuana use; mouse model; neuroAIDS; neurocognitive disorder; neuroinflammation; neuronal excitability; neuropathology; novel; preclinical study; receptor; response; success; tandem mass spectrometry

Project Summary In the context of human immunodeficiency virus (HIV) infection, cannabis use is an important topic and is the most commonly used drug among people living with HIV (PWH), despite the success of combined antiretroviral therapy (cART). Cannabis use is known to alleviate common symptoms and complications in PWH on cART, including chronic pain, nausea, and anxiety/depression, but contradicting results have been reported for cannabis use on cognitive performance. As cART does not cross the blood brain barrier the prevalence of mild to moderate forms of cognitive impairments, known as HIV-1-associated neurocognitive disorders (HAND), remain high (50%). Brain regions that are specifically vulnerable to HIV include the frontostriatal system, involving deficits in learning, memory and reward-related behaviors. HIV-1 proteins released by HIV-1 infected cells, including the transactivator of transcription (Tat) protein, are known to play a major role in the underlying HAND neuropathology, including lasting changes in neuronal excitability, inflammatory processes, and synaptic communication. Nevertheless, it is not clear how neuronal activity is altered by HIV Tat during learning while an animal is performing a behavioral task, i.e in the context of reward. With the establishment of in vivo calcium imaging, it has been shown that dorsomedial prefrontal cortex neurons that project to the nucleus accumbens (PFC-NAc) increase their neuronal activity after successful appetitive discrimination learning. The goal of the proposed study is to determine cannabis-induced changes on neuronal activity in a neuroHIV mouse model during performance of a behavioral reward-related learning task and its relation to the endocannabinoid (eCB) system. To achieve this goal, we will make use of a well-established mouse model of neuroHIV and propose two aims. Aim 1 will determine how CBD:THC ratios alter prefrontal neuronal activity in HIV Tat transgenic male and female mice performing a reward-related learning task. Animals will undergo viral injections to visualize in vivo calcium activity in PFC-NAc neurons and drug administration starts with behavioral assessment. Head-fixed mice will be trained to perform a Pavlovian conditioning task, that presents reward-predictive cues, and tests animal’s ability to learn to discriminate between a conditioned stimulus that predicts sucrose reward (CS+) but not the other (CS-). Neuronal activity will be recorded throughout training and compared before and after learning. Aim 2 will determine the effect of different CBD:THC ratios (1:15, 1:1, 15:1) on the eCB system and pathology in relation to appetitive discrimination learning in HIV Tat transgenic mice. Immunohistochemistry will be used to assess cell-specific localization of cannabinoid-like receptors (CB1R, CB2R, GPR55) and catabolic enzymes (FAAH, MAGL) on neurons, astrocytes, and microglia in the dorsomedial PFC and NAc; (non-)eCB ligands (e.g. AEA, 2-AG, AA, OEA, PEA) will be quantified using ultraperformance liquid chromatography- tandem mass spectrometry. This work will advance the understanding about how alterations in neuronal activity, inflammation, and cannabis-induced changes in the eCB system contribute to HIV-related symptoms.

项目摘要 在人类免疫缺陷病毒（HIV）感染的背景下，大麻使用是一个重要的话题，是 在艾滋病毒（PWH）的患者中，最常用的药物是抗逆转录病毒的成功的成功 治疗（购物车）。已知大麻的使用是为了减轻CART上PWH的常见症状和并发症， 包括慢性疼痛，恶心和动画/抑郁症，但据报道了矛盾的结果 大麻在认知表现上使用。由于购物车不越过血脑屏障，因此中期的患病率 对中等形式的认知障碍，称为HIV-1相关的神经认知障碍（手）， 保持高（50％）。特别容易受到艾滋病毒的大脑区域包括额叶系统， HIV-1蛋白被HIV-1感染 已知细胞，包括转录蛋白（TAT）蛋白的细胞在基础中起主要作用 手动神经病理学，包括神经元令人兴奋，炎症过程和突触的持久变化 沟通。然而，尚不清楚在学习过程中如何改变艾滋病毒的神经元活动 动物正在执行行为任务，即在奖励的背景下。建立体内钙 成像，已经显示出向伏拟核的背额额叶前额叶皮层神经元 （PFC-NAC）在成功欣赏歧视学习后增加其神经元活动。目标的目标 拟议的研究是确定在神经hiv小鼠模型中大麻诱导的神经元活性变化的变化 在执行与行为奖励相关的学习任务及其与内源性大麻素（ECB）的关系中 系统。为了实现这一目标，我们将利用建立完善的Neurohiv的鼠标模型和提案两个 目标。 AIM 1将确定CBD：THC比如何改变HIV TAT转基因男性和 雌性老鼠执行与奖励相关的学习任务。动物将接受病毒注射以可视化体内 PFC-NAC神经元和药物管理中的钙活性始于行为评估。头固定 小鼠将接受培训以执行帕夫洛维亚调节任务，提出奖励预测性线索和测试 动物学会区分有条件的刺激的能力，该刺激预测蔗糖奖励（CS+）但 不是另一个（CS-）。神经元活动将在整个训练中记录，并在之前和之后进行比较 学习。 AIM 2将确定不同CBD的影响：THC比（1：15，1：1，15：1）对欧洲央行系统的影响 与HIV TAT转基因小鼠中的食欲歧视学习有关的病理学。免疫组织化学将 用于评估大麻素样受体（CB1R，CB2R，GPR55）和分解代谢的细胞特异性定位 背侧PFC和NAC中神经元，星形胶质细胞和小胶质细胞上的酶（FAAH，MAGL）； （非）欧洲央行 配体（例如AEA，2-AG，AA，OEA，PEA）将使用超强化液相色谱进行定量 串联质谱。这项工作将促进人们对神经元活动的改变的理解， 炎症和大麻诱导的欧洲央行系统变化有助于与HIV相关的符号。