Alterations of Dopamine-Dependent Behaviors and Inflammation by HIV Proteins

描述(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，大约三分之一的HIV/AIDS患者会发生神经系统并发症。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，并与艾滋病毒特定蛋白存在引起的中枢神经系统慢性炎症反应有关。特别是，由于炎性介质的毒性水平，大脑中多巴胺能神经元含量高的区域容易受到损害和干扰。 以下实验旨在评估多巴胺在HIV-1转基因大鼠中的行为。这只大鼠表达与艾滋病毒有关的9个基因中的7个。它的用途之一是被提议作为暴露于HIV蛋白导致的神经缺陷的模型。这些实验将检验长期接触艾滋病毒蛋白对多巴胺和相关大脑区域介导的行为的影响。特定目标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。《特定目标2》将研究吗啡止痛作用的全部时间进程和剂量反应曲线，以及对热板刺激的基线疼痛反应。预计这些大鼠将表现出运动功能下降和对吗啡的止痛反应减弱，初步数据支持这些假设。此外，特殊目标3将通过分析大脑中特定的多巴胺能神经元丰富区域的炎症标志，如白细胞介素1β(IL-1β)、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)，来研究这些行为的神经生物学机制，方法是使用实时逆转录聚合酶链式反应(RT-PCR)来测量信使RNA(MRNA)，并使用酶联免疫吸附分析(ELISA)来测量蛋白质。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，以确定中枢神经系统的易感性是否随着暴露于HIV蛋白的时间而增加。 这些实验将评估慢性HIV蛋白暴露对中枢神经系统多巴胺能系统的影响。对多巴胺介导的行为变化的表征将使更多的研究人员能够研究新的疗法来治疗这些毁灭性的影响。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.