Alterations of Dopamine-Dependent Behaviors and Inflammation by HIV Proteins

描述（由申请人提供）：神经系统并发症在人类免疫缺陷病毒（HIV）/获得性免疫缺陷综合征（AIDS）患者中很常见，约占所有HIV/AIDS患者的三分之一。这些并发症包括神经性疼痛、运动功能障碍、认知障碍和行为异常，并且与由HIV的特定蛋白质的存在引起的中枢神经系统中的慢性炎症反应相关。特别是，多巴胺能神经元含量高的大脑区域由于炎症介质的毒性水平而易于受损和破坏。 设计以下实验以评价HIV-1转基因大鼠中多巴胺介导的行为。这只老鼠表达了与HIV有关的9个基因中的7个。其用途之一已被提议作为暴露于HIV蛋白质引起的神经缺陷的模型。这些实验将研究长期暴露于HIV蛋白对多巴胺和相关脑区介导的行为的影响。具体目标1将检查HIV-1转基因大鼠的运动功能，包括握力、步态和运动活动。具体目标2将检查吗啡镇痛作用的全时程和剂量反应曲线，以及对热板刺激的基线疼痛反应。预计这些大鼠将显示出运动功能降低和对吗啡的镇痛反应减弱，初步数据支持这些假设。此外，《特定目标3》将通过分析大脑中富含多巴胺能神经元的特定区域的炎症标志物，如白细胞介素-1 β（IL-1 β），来研究这些行为的神经生物学机制。）、肿瘤坏死因子-α（TNF-α）、单核细胞趋化蛋白-1（MCP-1）和诱导型一氧化氮合酶（iNOS），通过使用实时逆转录酶聚合酶链反应（RT-PCR）测量信使RNA（mRNA）和酶联免疫吸附测定（ELISA）测量蛋白质。为了充分了解HIV蛋白的累积作用，将对这些大鼠进行为期13个月的检查，以确定CNS脆弱性是否随暴露于HIV蛋白的时间增加而增加。 这些实验将评估慢性HIV蛋白暴露对中枢神经系统中多巴胺能系统的影响。多巴胺介导的行为改变的特征将允许更多的研究人员研究新的治疗方法来治疗这些破坏性的影响。此外，确定中枢神经系统中多巴胺能系统的神经炎症机制将为治疗HIV/AIDS患者常见的神经病变和运动功能障碍的药物提供新的靶点。

DESCRIPTION (provided by applicant): Neurological complications are common in human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) patients, occurring in approximately a third of all HIV/AIDS patients. These complications include neuropathic pain, motor dysfunctions, cognitive impairments, and behavioral abnormalities, and are associated with a chronic inflammatory response in the central nervous system caused by the presence of specific proteins of HIV. In particular, areas of the brain high in dopaminergic neurons are prone to damage and disruption due to toxic levels of inflammatory mediators. The following experiments are designed to evaluate dopamine-mediated behaviors in an HIV-1 transgenic rat. This rat expresses 7 of the 9 genes involved in HIV. One of its uses has been proposed as a model for neurological deficits induced by exposure to HIV proteins. These experiments will examine the effects of chronic exposure to HIV proteins on behaviors mediated by dopamine and associated brain regions. Specific Aim 1 will examine motor functions in the HIV-1 transgenic rats, including grip strength, gait, and locomotor activity. Specific Aim 2 will examine a full time course and dose response curve to analgesic effects of morphine, as well as baseline pain responses to a hot plate stimulus. It is expected that these rats will show reduced motor functions and diminished analgesic responses to morphine, and preliminary data supports these hypotheses. Additionally, Specific Aim 3 will examine the neurobiological mechanisms of these behaviors by analyzing specific dopaminergic neuron-rich areas of the brain for markers of inflammation such as interleukin-1beta (IL-1¿), tumor necrosis factor-alpha (TNF-a), monocyte chemotactic protein-1 (MCP-1) and inducible nitric oxide synthase (iNOS) through the use of real-time reverse transcriptase polymerase chain reaction (RT-PCR) to measure messenger RNA (mRNA) and enzyme-linked immunosorbent assays (ELISAs) to measure protein. To fully understand the cumulative actions of HIV proteins, these rats will be examined across a period of 13 months to determine if CNS vulnerability is increased with length of exposure to HIV proteins. These experiments will evaluate the effects of chronic HIV protein exposure on dopaminergic systems in the central nervous system. The characterization of the alterations in dopamine-mediated behaviors will allow more investigators to research novel therapeutics to treat these devastating effects. In addition, identifying a neuroinflammatory mechanism in the dopaminergic systems in the central nervous system will provide a new target for drugs to treat neuropathy and motor dysfunctions often seen in HIV/AIDS patients.