Automated Microscopy and Image Analysis for Androgen Receptor Function
Automated Microscopy and Image Analysis for Androgen Receptor Function
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DOI：
10.1007/978-1-61779-243-4_18
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发表时间：
2011-01-01
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影响因子：
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通讯作者：
Mancini, Michael A.
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文献类型：
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作者：
Hartig, Sean M.;Newberg, Justin Y.;Mancini, Michael A.
系统级方法已经出现，依赖于分析，基于显微镜的技术，用于发现新的药物靶标和驱动AR信号传导，转录活性和配体独立性的机制。单细胞行为可以通过高通量显微镜方法通过分析内源性蛋白质水平和生物传感器细胞系的定位或创建来定量，所述生物传感器细胞系可以同时检测对已知和未知雄激素刺激的急性和潜伏反应。细胞成像和分析方案可以自动化以发现核易位、报告基因活性、核输出和亚核转录事件的激动剂/拮抗剂反应窗口，从而促进获得通过经典生物化学方法固有地不可用的多重模型系统。在本章中，我们将重点介绍开发、进行和分析高通量筛选以识别AR信号效应子所需的关键步骤。
Systems-level approaches have emerged that rely on analytical, microscopy-based technology for the discovery of novel drug targets and the mechanisms driving AR signaling, transcriptional activity, and ligand independence. Single cell behavior can be quantified by high-throughput microscopy methods through analysis of endogenous protein levels and localization or creation of biosensor cell lines that can simultaneously detect both acute and latent responses to known and unknown androgenic stimuli. The cell imaging and analytical protocols can be automated to discover agonist/antagonist response windows for nuclear translocation, reporter gene activity, nuclear export, and subnuclear transcription events, facilitating access to a multiplex model system that is inherently unavailable through classic biochemical approaches. In this chapter, we highlight the key steps needed for developing, conducting, and analyzing high-throughput screens to identify effectors of AR signaling.