Center for Aging and Regeneration, Millennium Nucleus in Regenerative Biology, Faculty of Biological Sciences, Pontificia Universidad Católica de Chile, Santiago, Chile.
Nat Protoc. 2017 Feb;12(2):372-389. doi: 10.1038/nprot.2016.177. Epub 2017 Jan 19.
Here we present a protocol for the husbandry of Xenopus laevis tadpoles and froglets, and procedures to study spinal cord regeneration. This includes methods to induce spinal cord injury (SCI); DNA and morpholino electroporation for genetic studies; in vivo imaging for cell analysis; a swimming test to measure functional recovery; and a convenient model for screening for new compounds that promote neural regeneration. These protocols establish X. laevis as a unique model organism for understanding spinal cord regeneration by comparing regenerative and nonregenerative stages. This protocol can be used to understand the molecular and cellular mechanisms involved in nervous system regeneration, including neural stem and progenitor cell (NSPC) proliferation and neurogenesis, extrinsic and intrinsic mechanisms involved in axon regeneration, glial response and scar formation, and trophic factors. For experienced personnel, husbandry takes 1-2 months; SCI can be achieved in 5-15 min; and swimming recovery takes 20-30 d.
我们在此介绍了爪蟾蝌蚪和蛙的饲养方案，以及研究脊髓再生的程序。这包括诱导脊髓损伤（SCI）的方法；用于遗传研究的 DNA 和 morpholino 电穿孔；用于细胞分析的体内成像；用于测量功能恢复的游泳测试；以及筛选促进神经再生的新化合物的便利模型。这些方案通过比较再生和非再生阶段，将 X. laevis 确立为研究脊髓再生的独特模式生物。该方案可用于了解神经系统再生所涉及的分子和细胞机制，包括神经干细胞和祖细胞（NSPC）的增殖和神经发生、轴突再生所涉及的外在和内在机制、神经胶质反应和疤痕形成以及营养因子。对于有经验的人员，饲养需要 1-2 个月；SCI 可以在 5-15 分钟内完成；游泳恢复需要 20-30 天。
