Isensee Jörg, Hucho Tim
Experimental Anesthesiology and Pain Research, Department of Anesthesiology and Intensive Care Medicine, University Hospital Cologne, University of Cologne, Cologne, Germany.
Methods Mol Biol. 2019;1987:111-124. doi: 10.1007/978-1-4939-9446-5_8.
Studying TRP channel expressing nociceptors requires the identification of the respective subpopulations as well as the quantification of dynamic cellular events. However, the heterogeneity of sensory neurons and associated nonneuronal cells demands the analysis of large numbers of cells to reflect the distribution of entire populations. Here we report a detailed workflow how to apply high-content screening (HCS) microscopy to signaling events in TRPV1-positive neurons as well as an approach to use the selective elimination of TRPV1 positive cells from dissociated rat sensory ganglia as base for transcriptomic analysis of TRPV1-positive cells and/or as control for TRPV1 antibody specificity.
研究表达TRP通道的伤害感受器需要识别各自的亚群以及对动态细胞事件进行定量分析。然而，感觉神经元和相关非神经元细胞的异质性要求对大量细胞进行分析，以反映整个群体的分布情况。在此，我们报告一种详细的工作流程，即如何将高内涵筛选（HCS）显微镜应用于TRPV1阳性神经元中的信号事件，以及一种利用从解离的大鼠感觉神经节中选择性消除TRPV1阳性细胞的方法，作为TRPV1阳性细胞转录组分析的基础和/或作为TRPV1抗体特异性的对照。
