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（艾滋病毒）感染的情况下，大麻使用是一个重要的议题， 尽管联合抗逆转录病毒治疗取得了成功， 治疗（cART）已知大麻的使用可以减轻接受cART的PWH患者的常见症状和并发症， 包括慢性疼痛、恶心和焦虑/抑郁，但据报道， 大麻对认知能力的影响由于cART不能穿过血脑屏障，因此轻度的 中度认知障碍，称为HIV-1相关神经认知障碍（HAND）， 高（50%）。特别容易感染艾滋病毒的大脑区域包括额纹状体系统， 包括学习记忆和奖励相关行为的缺陷。HIV-1感染者释放的HIV-1蛋白 细胞，包括转录的反式激活因子（达特）蛋白，已知在潜在的 手部神经病理学，包括神经元兴奋性、炎症过程和突触 通信尽管如此，目前尚不清楚艾滋病毒达特在学习期间如何改变神经元活动，而在学习期间则如何改变神经元活动 动物正在执行一项行为任务，即在奖励的背景下。随着体内钙离子的建立 通过成像，已经表明投射到丘脑核的背内侧前额叶皮层神经元 （PFC-NAc）增加成功的食欲辨别学习后的神经元活动。的目标 拟议的研究是在神经HIV小鼠模型中确定大麻诱导的神经元活性变化 行为奖励相关学习任务的执行过程及其与内源性大麻素（eCB）的关系 系统为了实现这一目标，我们将利用一个完善的神经HIV小鼠模型，并提出两个 目标。目的1将确定CBD：THC比例如何改变HIV达特转基因男性的前额神经元活动， 雌性小鼠执行奖励相关的学习任务。动物将接受病毒注射以在体内可视化 PFC-NAc神经元中的钙活性和药物施用从行为评估开始。头部固定 老鼠将被训练来执行巴甫洛夫条件反射任务，该任务提供奖励预测线索， 动物学习区分预测蔗糖奖励（CS+）的条件刺激， 而不是另一个（CS-）。在整个训练过程中记录神经元活动，并在训练前后进行比较。 学习目标2将确定不同CBD：THC比例（1：15、1：1、15：1）对eCB系统的影响， 与HIV达特转基因小鼠的食欲辨别学习有关的病理学。免疫组织化学将 用于评估大麻素样受体（CB 1 R，CB 2 R，GPR 55）和分解代谢的细胞特异性定位 背内侧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）eCB 将使用超高效液相色谱法对配体（例如AEA、2-AG、AA、OEA、PEA）进行定量。 串联质谱法这项工作将促进对神经元活动如何改变的理解， 炎症和大麻诱导的eCB系统变化有助于HIV相关症状。