Li Zhuyin, Yan Yongping, Powers Elaine A, Ying Xiaoyou, Janjua Khurram, Garyantes Tina, Baron Bruce
Lead Discovery Technology, Lead Generation, Aventis Pharmaceutical, Bridgewater, NJ 08807, USA.
J Biomol Screen. 2003 Oct;8(5):489-99. doi: 10.1177/1087057103257309.
Gap junctions coordinate electrical signals and facilitate metabolic synchronization between cells. In this study, the authors have developed a novel assay for the identification of gap junction blockers using fluorescence microscopy imaging-based high-content screening technology. In the assay, the communication between neighboring cells through gap junctions was measured by following the redistribution of a fluorescent marker. The movement of calcein dye from dye-loaded donor cells to dye-free acceptor cells through gap junctions overexpressed on cell surface membranes was monitored using automated fluorescence microscopy imaging in a high-throughput compatible format. The fluorescence imaging technology consisted of automated focusing, image acquisition, image processing, and data mining. The authors have successfully performed a high-throughput screening of a 486,000- compound program with this assay, and they were able to identify false positives without additional experiments. Selective and pharmacologically interesting compounds were identified for further optimization.
