Live Imaging of Neural Crest and Pigment Cells and Transient Transgenic Manipulation of Gene Activity

  • Reyna Deeya Ballim
  • Yusuke Nagao
  • Robert Neil KelshEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1976)


Neural crest cells are a highly multipotent and migratory cell type that are important for adult pigment pattern formation, cellular homeostasis, and regeneration. The optical transparency and accessibility of fish embryos makes 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. We describe key protocols for the analysis of neural crest development in zebrafish and medaka, including live imaging of neural crest cells and differentiating pigment cells and transient transgenesis assays that can be used to manipulate neural crest development.

Key words

Neural crest Pigment cells Zebrafish Medaka Live imaging Melanocyte Xanthophore Leucophore Iridophore Transient transgenesis Tol2 



This work was supported by BBSRC grant BB/L00769X/1 (R.N.K.), Marie Sklodowska-Curie grant No. 661616 (R.D.B.), and Japanese Society for the Promotion of Research Program for Advancing Strategic International Networks to Accelerate the Circulation of Talented Researchers S2602 (Y.N.) and Overseas Research Fellowships No. 598 (Y.N.).


  1. 1.
    Kelsh RN, Brand M, Jiang YJ et al (1996) Zebrafish pigmentation mutations and the processes of neural crest development. Development 123:369–389PubMedGoogle Scholar
  2. 2.
    Lamoreux LM, Kelsh RN, Wakamatsu Y et al (2005) Pigment pattern formation in the medaka embryo. Pigment Cell Res 18:64–73CrossRefGoogle Scholar
  3. 3.
    Schartl M, Larue L, Goda M et al (2016) What is a vertebrate pigment cell? Pigment Cell Melanoma Res 29:8–14CrossRefGoogle Scholar
  4. 4.
    Westerfield M (2000) The Zebrafish book. A guide for the laboratory use of zebrafish (Danio rerio). University of Oregon Press, Eugene, OregonGoogle Scholar
  5. 5.
    Rosen JN, Sweeney MF, Mably JD (2009) Microinjection of zebrafish embryos to analyze gene function. J Vis Exp (25):1115Google Scholar
  6. 6.
    Kawakami K, Imanaka K, Itoh M, Taira M (2004) Excision of the Tol2 transposable element of the medaka fish Oryzias latipes in Xenopus laevis and Xenopus tropicalis. Gene 338:93–98CrossRefGoogle Scholar
  7. 7.
    Lee EM, Yuan T, Ballim RD, Nguyen K, Kelsh RN, Medeiros DM, McCauley DW (2016) Functional constraints on SoxE proteins in neural crest development: the importance of differential expression for evolution of protein activity. Dev Biol 418:166–178CrossRefGoogle Scholar
  8. 8.
    Elworthy S, Lister JA, Carney TJ, Raible DW, Kelsh RN (2003) Transcriptional regulation of mitfa accounts for the sox10 requirement in zebrafish melanophore development. Development 130:2809–2818CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Reyna Deeya Ballim
    • 1
  • Yusuke Nagao
    • 1
  • Robert Neil Kelsh
    • 1
    Email author
  1. 1.Department of Biology and BiochemistryUniversity of BathBathUK

Personalised recommendations