PUBLICATION

Generation of Naïve Blastoderm Explants from Zebrafish Embryos

Authors
Alaniz Emig, A., Williams, M.L.K.
ID
ZDB-PUB-210817-11
Date
2021
Source
Journal of visualized experiments : JoVE   (173): (Journal)
Registered Authors
Williams, Margot
Keywords
none
MeSH Terms
  • Animals
  • Blastoderm*/metabolism
  • Body Patterning
  • Gene Expression Regulation, Developmental
  • Morphogenesis
  • Zebrafish*/genetics
  • Zebrafish*/metabolism
  • Zebrafish Proteins/metabolism
PubMed
34398141 Full text @ J. Vis. Exp.
Abstract
Due to their optical clarity and rapid development, zebrafish embryos are an excellent system for examining cell behaviors and developmental processes. However, because of the complexity and redundancy of embryonic signals, it can be challenging to discern the complete role of any single signal during early embryogenesis. By explanting the animal region of the zebrafish blastoderm, relatively naïve clusters of embryonic cells are generated that can be easily cultured and manipulated ex vivo. By introducing a gene of interest by RNA injection before explantation, one can assess the effect of this molecule on gene expression, cell behaviors, and other developmental processes in relative isolation. Furthermore, cells from embryos of different genotypes or conditions can be combined in a single chimeric explant to examine cell/tissue interactions and tissue-specific gene functions. This article provides instructions for generating zebrafish blastoderm explants and demonstrates that a single signaling molecule - a Nodal ligand - is sufficient to induce germ layer formation and extension morphogenesis in otherwise naïve embryonic tissues. Due to their ability to recapitulate embryonic cell behaviors, morphogen gradients, and gene expression patterns in a simplified ex vivo system, these explants are anticipated to be of great utility to many zebrafish researchers.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping