Direct and indirect striatal pathway selective vulnerability to Tat and morphine

直接和间接纹状体通路对 Tat 和吗啡的选择性脆弱性

• 批准号: 8846847
• 负责人: Christina Joanne Schier
• 金额: $ 4.97万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-02-05 至 2016-02-04
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8846847
• 关键词: Address; Adenosine; Affect; Automobile Driving; Basal Ganglia; Behavioral; Biological Assay; Brain region; Calcium; Cells; Cessation of life; Cognition; Cognitive; Cognitive deficits; Comorbidity; Complex; Corpus striatum structure; Dendritic Spines; Dopamine; Doxycycline; Drug abuse; Dynorphins; Encephalitis; Enkephalins; Evaluation; Functional disorder; Genetic Transcription; Globus Pallidus; Goals; HIV; HIV Infections; HIV-1; Human; Hybrids; Image; Implant; In Vitro; Individual; Infection; Injection of therapeutic agent; Investigation; Lead; Learning; Length; Locomotion; Longevity; Mediating; Messenger RNA; Microscopy; Midbrain structure; Modeling; Morphine; Motor; Movement; Mus; Muscarinic M1 Receptor; Muscarinics; Nerve Degeneration; Neuraxis; Neuroglia; Neurons; Opioid; Opioid Receptor; Pathology; Pathway interactions; Patients; Pharmaceutical Preparations; Population; Prevalence; Property; Proteins; Role; Sampling; Societies; Structure; Subgroup; Substance P; Substantia nigra structure; Surface; Symptoms; Testing; Time; Trans-Activators; Transgenic Mice; Vertebral column; antiretroviral therapy; base; biocytin; design; human CREB1 protein; improved; in vivo; in vivo Model; insight; motor deficit; motor impairment; mouse model; neuroAIDS; neurocognitive disorder; neuron loss; neurotoxic; neurotoxicity; novel; opioid abuse; patch clamp; promoter; protein expression; public health relevance; receptor; reconstruction; selective expression; temporal measurement

 DESCRIPTION (provided by applicant): The advent of combined antiretroviral therapy (cART) has substantially extended the lifespan of HIV-1 patients; yet, the prevalence of HIV-1 associated neurocognitive disorders (HAND), which can be exacerbated by co-morbid opioid drug abuse, overall has not changed. Many HAND symptoms are thought to be caused by the selective degeneration of particular brain regions, such as the striatum, during HIV-1 infection. The striatum is comprised of approximately 90-95% medium spiny neurons (MSNs). The MSN population is heterogeneous, and can be divided in subtypes based on their expression of different combinations of surface markers and proteins, such as dopamine 1 and 2 (D1, D2), adenosine A2A, and muscarinic M4 receptors, as well as, substance P, dynorphin, and enkephalin, with D1- and D2-expressing MSN as the main subtypes. Assessment of MSN subtype vulnerability to HIV-1/opioid-use could enhance our understanding of how HIV-1 and co-morbid opioid use lead to HAND symptoms, and suggest refined preventative and functional treatments. The proposed studies will take advantage of dopamine Drd1 (D1) or Drd2 (D2) -receptor promoter-driven tdTomato- or EFGP-expressing mouse lines, respectively, to identify D1- and D2-receptor- expressing MSNs. These mouse lines, along with wild-type controls, will be used to investigate lethal and sub-lethal effects of the HIV-1 protein transactivator of transcription (Tat) and opioids on the subtypes of MSNs using both in vitro and in vivo models. Primary mixed cortical-striatal, neuron-glia cultures will be employed to determine the selective direct and glial-induced indirect neurotoxic and synaptodendritic degenerative effects of Tat and morphine on these subtypes. We will then use a D1-tdTomato- and D2-EGFP-expressing mouse line crossed with a doxycycline-inducible Tat-expressing mouse line to explore if previously noted Tat- and/or morphine-induced motor disturbances are selectively mediated through synaptodendritic degeneration, and/or electrophysiological changes in a particular subset of MSNs. This will be accomplished using a battery of mouse-motor assays, whole-cell patch clamp, and three-dimensional reconstruction of D1- and D2-expressing MSNs. Taken together, these experimental results will show if a subgroup of MSNs is driving the previously noted Tat- and morphine-induced lethal and synaptodendritic degenerative effects, and their correlation with motor impairments. The combination of in vitro approaches, which allow for precise control of experimental conditions, and in vivo tests, which will show a functional/morphological correlation, will provide the first described insights into the selective vulnerability of MSN subtypes to HIV-1 and/or opioids.

 描述（由申请人提供）：联合抗逆转录病毒治疗（cART）的出现大大延长了HIV-1患者的寿命;然而，HIV-1相关神经认知障碍（HAND）的患病率总体上没有改变，HAND可因合并阿片类药物滥用而加重。许多HAND症状被认为是由HIV-1感染期间特定大脑区域（如纹状体）的选择性变性引起的。纹状体由大约90-95%的中型多刺神经元（MSN）组成。MSN群体是异质性的，并且可以基于其表面标志物和蛋白质的不同组合的表达而分为亚型，所述表面标志物和蛋白质例如多巴胺1和2（D1、D2）、腺苷A2 A和毒蕈碱M4受体，以及P物质、强啡肽和脑啡肽，其中表达D1和D2的MSN为主要亚型。评估MSN亚型对HIV-1/阿片类药物使用的脆弱性可以增强我们对HIV-1和合并阿片类药物使用如何导致HAND症状的理解，并提出完善的预防和功能性治疗。拟议的研究将分别利用多巴胺Drd 1（D1）或Drd 2（D2）受体启动子驱动的tdTomato或EFGP表达小鼠系，以鉴定D1和D2受体表达MSN。这些小鼠系，连同野生型对照沿着，将用于使用体外和体内模型研究HIV-1蛋白转录反式激活因子（达特）和阿片类药物对MSN亚型的致死和亚致死作用。原代混合的皮质-纹状体，神经元-胶质细胞培养物将被用来确定达特和吗啡对这些亚型的选择性直接和胶质诱导的间接神经毒性和突触树突变性作用。然后，我们将使用D1-tdTomato-和D2-EGFP-表达小鼠系与多西环素诱导的Tat-表达小鼠系杂交，以探索先前指出的达特-和/或吗啡诱导的运动障碍是否通过突触树突状细胞变性和/或特定MSN亚群的电生理变化选择性介导。这将使用一组小鼠运动试验、全细胞膜片钳和表达D1和D2的MSN的三维重建来完成。总之，这些实验结果将表明，如果一个亚组的MSN驱动先前指出的达特和吗啡诱导的致死和突触树突变性效应，以及它们与运动障碍的相关性。允许精确控制实验条件的体外方法和将显示功能/形态学相关性的体内测试的组合将提供对MSN亚型对HIV-1和/或阿片类药物的选择性脆弱性的首次描述的见解。