Instituto de Biología Molecular de Barcelona, CSIC, Parc Científic de Barcelona, Barcelona, Spain.
Centre de Biologie du Développement (CBD), Centre de Biologie Intégrative (CBI), Université de Toulouse, CNRS, UPS, Toulouse, France.
Methods Mol Biol. 2021;2179:183-197. doi: 10.1007/978-1-0716-0779-4_16.
The neural tube in amniotic embryos forms as a result of two consecutive events along the anteroposterior axis, referred to as primary and secondary neurulation (PN and SN). While PN involves the invagination of a sheet of epithelial cells, SN shapes the caudal neural tube through the mesenchymal-to-epithelial transition (MET) of neuromesodermal progenitors, followed by cavitation of the medullary cord. The technical difficulties in studying SN mainly involve the challenge of labeling and manipulating SN cells in vivo. Here we describe a new method to follow MET during SN in the chick embryo, combining early in ovo chick electroporation with in vivo time-lapse imaging. This procedure allows the cells undergoing SN to be manipulated in order to investigate the MET process, permitting their cell dynamics to be followed in vivo.
羊膜胚胎中的神经管通过沿着前后轴的两个连续事件形成，这两个事件被称为初级和次级神经形成（PN 和 SN）。虽然 PN 涉及上皮细胞片的内陷，但 SN 通过神经中胚层祖细胞的间质到上皮过渡（MET）来塑造尾侧神经管，随后髓索空化。研究 SN 的技术困难主要涉及在体内标记和操作 SN 细胞的挑战。在这里，我们描述了一种在鸡胚中跟踪 SN 期间 MET 的新方法，将早期鸡胚电穿孔与体内延时成像相结合。该程序允许对经历 SN 的细胞进行操作，以研究 MET 过程，从而可以在体内跟踪它们的细胞动力学。
