Abstract
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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Thisse B, Thisee C (2004) Fast release clones: a hgh throughput expression analysis. ZFIN Direct Data Submission
Gross-Thebing T, Paksa A, Raz E (2014) Simultaneous high-resolution detection of multiple transcripts combined with localization of proteins in whole-mount embryos. BMC Biol 12:55
Wang F, Flanagan J, Su N, Wang LC, Bui S, Nielson A, Wu X, Vo HT, Ma XJ, Luo Y (2012) RNAscope: a novel in situ RNA analysis platform for formalin-fixed, paraffin-embedded tissues. J Mol Diagn 14:22–29
Acknowledgments
This work was supported by Sultan Qaboos University Scholarship (R.A.J.), BBSRC SWBio DTP (K.P.), the Japanese Society for the Promotion of Research Program for Advancing Strategic International Networks to Accelerate the Circulation of Talented Researchers S2602 (Y.N.) and CONACyT grant 329640/384511 (K.C.-S.), and BBSRC grant BB/L00769X/1 and MRC grant MR/J001457/1 (R.N.K.). We gratefully acknowledge expert guidance on RNAscope from Prof. E. Raz and Dr. A. Paksa (University of Muenster).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Petratou, K., Camargo-Sosa, K., Al Jabri, R., Nagao, Y., Kelsh, R.N. (2019). Neural Crest Methodologies in Zebrafish and Medaka. In: Schwarz, Q., Wiszniak, S. (eds) Neural Crest Cells. Methods in Molecular Biology, vol 1976. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9412-0_13
Download citation
DOI: https://doi.org/10.1007/978-1-4939-9412-0_13
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-9411-3
Online ISBN: 978-1-4939-9412-0
eBook Packages: Springer Protocols