Department of Cell Biology, Duke University Medical Center, Durham, NC, USA.
Duke Regeneration Center, Duke University, Durham, NC, USA.
Methods Mol Biol. 2024;2707:185-204. doi: 10.1007/978-1-0716-3401-1_12.
In regeneration, a damaged body part grows back to its original form. Understanding the mechanisms and physical principles underlying this process has been limited by the difficulties of visualizing cell signals and behaviors in regeneration. Zebrafish scales are emerging as a model system to investigate morphogenesis during vertebrate regeneration using quantitative live imaging. Scales are millimeter-sized dermal bone disks forming a skeletal armor on the body of the fish. The scale bone is deposited by an adjacent monolayer of osteoblasts that, after scale loss, regenerates in about 2 weeks. This intriguing regenerative process is accessible to live confocal microscopy, quantifications, and mathematical modeling. Here, I describe methods to image scale regeneration live, tissue-wide and at sub-cellular resolution. Furthermore, I describe methods to process the resulting images and quantify cell, tissue, and signal dynamics.
在再生过程中,受损的身体部位会恢复到原来的形态。由于难以可视化细胞信号和行为,因此对这一过程背后的机制和物理原理的理解受到了限制。斑马鱼鳞片正在成为一种新兴的模型系统,可通过定量活体成像研究脊椎动物再生过程中的形态发生。鳞片是毫米大小的真皮骨盘,在鱼体上形成骨骼盔甲。鳞片骨由相邻的单层成骨细胞沉积,在鳞片丢失后,大约 2 周内再生。这种引人入胜的再生过程可以进行活体共聚焦显微镜观察、定量分析和数学建模。在这里,我描述了对鳞片再生进行活体、全组织和亚细胞分辨率成像的方法。此外,我还描述了处理这些图像并量化细胞、组织和信号动态的方法。