NEUROBIOLOGY

这是Masliah对Project的修改后的应用程序，已进行了广泛修改，以解决评审员的担忧：1）将我们的分析扩展到包括体视学和定量真实的时间PCR（qRT-PCR），2）扩展我们对星形胶质细胞的神经病理学和体外研究，以及3）结合新的初步数据以支持更创新的概念，即由HIV和HCV感染的细胞产生的干扰素（IFN）可能，与METH结合，增强可能促进神经变性的星形胶质细胞反应，并阻断神经营养因子如成纤维细胞生长因子（FGF）对钙结合蛋白（CB）的作用， 多巴胺能神经元具体而言，我们假设，CB-IR在新皮层和海马的损失将与认知缺陷，而在黑质窦系统的标记物的损失将与运动和认知缺陷。对于PPG的更新和与其他组件的合作，我们的目标是：1）阐明METH+，HIV+病例的脑中的区域神经元损伤和星形胶质细胞增生的模式，以及它们与HIVE的神经病理学标志物和IFN表达水平以及对皮质中间神经元和NS细胞选择性的营养因子的关系。2}为了确定使用METH的HIV感染病例的大脑中神经元损伤和星形胶质细胞增生的区域模式之间的关系，这些病例在HCV状态、特定脑脊液（CSF）生物标志物和行为改变方面各不相同。3)通过星形胶质细胞IFN-α的改变研究METH和HIV在体外的协同神经毒性作用 和IFI基因表达和神经元FGF/SHH信号传导。将在存在或不存在METH和FGF 2或FGFS和SHH的情况下，在暴露于HIV蛋白的神经元细胞系、原代神经元培养物和人胎脑聚集体中进行分子机制的剖析。这些研究将有助于进一步了解艾滋病毒感染的METH用户的认知和运动改变的细胞和分子机制，并为这些复杂的疾病开发新的治疗策略。

This is a revised application for Project by Masliah that has been extensively modified to address the reviewers' concerns by: 1) expanding our analysis to include stereology and quantitative real time PCR (qRT-PCR), 2) broadening our neuropathological and in vitro studies of astroglial cells, and 3) incorporating new preliminary data in support of the more innovative concept that interferons (IFNs) produced by HIV and HCV-infected cells might, in conjunction with METH, enhance astroglial responses that might promote neurodegeneration and block the effects of neurotrophic factors such as fibroblast growth factors (FGFs) on calbindin (CB) and dopaminergic neurons. Specifically, we hypothesize that loss of CB-IR in the neocortex and hippocampus will be associated with cognitive deficits, while loss of markers in the nigrostiatal system will correlate with motor and cognitive deficits. For the renewal and in collaboration with other components of the PPG our aims are: 1) To elucidate patterns of regional neuronal damage and astrogliosis in the brains of METH+, HIV+ cases and their relationship with neuropathological markers of HIVE and levels of IFN expression and trophic factors selective for cortical interneurons and NS cells. 2} To determine the relationship between regional patterns of neuronal injury and astrogliosis in the brains of METH-using HIV infected cases who vary in HCV status, specific cerebrospinal fluid (CSF) biomarkers and behavioral alterations. 3) To investigate the synergistic neurotoxic effects of METH and HIV in vitro via alterations in astroglial IFN-alpha and IFI gene expression and neuronal FGF/SHH signaling. Dissection of the molecular mechanisms will be performed in neuronal cell lines, primary neuronal cultures, and human fetal brain aggregates exposed to HIV proteins in the presence or absence of METH and FGF2, or FGFS and SHH. These studies will help advance understanding of the cellular and molecular mechanisms of cognitive and motor alterations in HIV infected METH users and in developing novel treatment strategies for these complex disorders.