State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, 610041, Sichuan, China.
Department of Biomedical Sciences, Texas A&M College of Dentistry, Dallas, 75246, TX, USA.
Sci Rep. 2018 Jul 5;8(1):10184. doi: 10.1038/s41598-018-28205-3.
Osteoclasts are multinucleated giant cells. Fusion is an essential element in the formation of osteoclasts. However, the exact cellular events and mechanisms remain largely unknown because of limited and insufficient methods for observing fusion process. In this work, a fluorescence reporter strategy was established to monitor osteoclast fusion. After fusing with cells expressing Cre recombinase, those cells with double fluorescence switch its expression from red to green fluorescent protein. The effect of RANKL and PTH on osteoclast fusion were both quantitatively and visually detected utilizing this strategy. Furthermore, a combination of this strategy with a technique of fluorescence-activated cell sorting revealed two different populations of fused osteoclasts, tdTomato+ GFP+ cells (TG cells) and GFP+ cells (G cells). The results argue for the potential of combining this technique with other bio-technologies to gain more information about osteoclast fusion. Overall, these data demonstrated that this visual fluorescence switch strategy is useful for further analysis of osteoclast fusion mechanisms.
破骨细胞是多核巨细胞。融合是破骨细胞形成的一个必要元素。然而，由于用于观察融合过程的方法有限且不足，确切的细胞事件和机制在很大程度上仍然未知。在这项工作中，建立了一种荧光报告策略来监测破骨细胞融合。与表达 Cre 重组酶的细胞融合后，那些具有双荧光的细胞将其表达从红色荧光蛋白切换为绿色荧光蛋白。利用该策略定量和直观地检测了 RANKL 和 PTH 对破骨细胞融合的影响。此外，该策略与荧光激活细胞分选技术的结合揭示了两种不同的融合破骨细胞群体，tdTomato+ GFP+ 细胞（TG 细胞）和 GFP+ 细胞（G 细胞）。结果表明，该技术与其他生物技术相结合具有获得更多关于破骨细胞融合信息的潜力。总的来说，这些数据表明，这种可视化荧光开关策略可用于进一步分析破骨细胞融合机制。
