Neural Crest Methodologies in Zebrafish and Medaka

Neural crest cells are highly multipotent and strongly migratory cells and generate adult neural crest stem cells with varied roles in cellular homeostasis and regeneration. The optical transparency and ready accessibility of fish embryos make them particularly well-suited to high-resolution analysis of neural crest development. However, the dispersive nature of these cells adds to the challenge of their study, requiring that they be identified using marker expression. We describe key protocols for the analysis of neural crest marker expression in zebrafish and medaka, including whole-mount in situ hybridization to detect mRNA using conventional chromogenic substrates and the more recent RNAscope which gives readily multiplexed fluorescent detection and immunofluorescent detection of antigens.