The State Key Laboratory of Cell Biology, Shanghai Institute of Biochemistry and Cell Biology, Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China.
The State Key Laboratory of Cell Biology, Shanghai Institute of Biochemistry and Cell Biology, Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China; School of Life Science and Technology, Shanghai Tech University, Shanghai 201210, China.
J Mol Cell Cardiol. 2020 Sep;146:60-68. doi: 10.1016/j.yjmcc.2020.07.002. Epub 2020 Jul 12.
Genetic lineage tracing has been widely used for studying in vivo cell fate plasticity during embryogenesis, tissue homeostasis, and disease development. Recent applications with multiple site-specific recombinases have been used in complex and sophisticated genetic fate mapping studies. However, the previous multicolor reporters for dual recombinases had limitations of precise in situ quantification of cell number, which is mainly due to the intermingling of cells in condensed tissues. Here, we generated a dual recombinase-mediated nuclear-localized GFP and tdTomato reporter line, which enables clear, simultaneous quantification of two distinct cell lineages in vivo. Combining this dual genetic reporter with Tbx18-Cre and Cdh5-Dre lines, which genetically trace epicardial and endothelial cells, respectively, we obtained high-resolution images for the anatomic distribution of the descendants of these two distinct cell lineages in the valve mesenchyme during development, remodeling, and maturation stages. This new dual genetic reporter is expected to facilitate fate tracing of two cell lineages and their objective quantification in vivo.
遗传谱系追踪已广泛用于研究胚胎发生、组织稳态和疾病发展过程中的体内细胞命运可塑性。最近，多种位点特异性重组酶的应用已用于复杂而精细的遗传命运映射研究。然而，以前用于双重组酶的多色报告器在精确原位定量细胞数量方面存在局限性，这主要是由于浓缩组织中细胞的混杂。在这里，我们生成了一种双重组酶介导的核定位 GFP 和 tdTomato 报告基因系，可在体内清楚、同时定量两种不同的细胞谱系。将这种双遗传报告器与 Tbx18-Cre 和 Cdh5-Dre 系结合，分别对心外膜细胞和内皮细胞进行基因追踪，我们获得了在发育、重塑和成熟阶段，这两种不同细胞谱系在瓣膜间质中的后代的高分辨率图像。这个新的双遗传报告器有望促进两个细胞谱系的命运追踪及其在体内的客观定量。
