Raymond Michael J, McCusker Catherine D
Brigham and Women's Hospital, Boston, MA 02115, USA.
Department of Biology, University of Massachusetts Boston, Boston, MA 02115, USA.
STAR Protoc. 2025 Jun 27;6(3):103920. doi: 10.1016/j.xpro.2025.103920.
A key question in limb regeneration is how axolotl limb cells become competent to specific types of regeneration signals. Here, we present the competency accessory limb model (CALM), a regenerative assay that assesses whether limb cells have achieved the ability to respond to patterning signals. We describe the steps for positioning a wound, deviating the nerve into a limb wound site, and injecting retinoic acid. We also detail the harvest and analyses in both anterior and posterior CALMs. For complete details on the use and execution of this protocol, please refer to Raymond et al..
肢体再生中的一个关键问题是蝾螈肢体细胞如何变得能够对特定类型的再生信号产生反应。在此,我们提出了能力附属肢体模型(CALM),这是一种再生分析方法,用于评估肢体细胞是否已获得对模式信号作出反应的能力。我们描述了伤口定位、将神经偏向肢体伤口部位以及注射视黄酸的步骤。我们还详细说明了前侧和后侧CALM中的收获和分析过程。有关此方案的使用和执行的完整详细信息,请参考雷蒙德等人的研究。