PUBLICATION

Generation of Ectopic Morphogen Gradients in the Zebrafish Blastula

Authors
de Olivera-Melo, M., Xu, P.F., Houssin, N., Thisse, B., Thisse, C.
ID
ZDB-PUB-181017-14
Date
2018
Source
Methods in molecular biology (Clifton, N.J.)   1863: 125-141 (Chapter)
Registered Authors
Houssin, Nathalie, Thisse, Bernard, Thisse, Christine, Xu, Peng-Fei
Keywords
Bead implantation, Blastula, Cell transplantation, Embryoid bodies, Gradient, Injection, Morphogen, Zebrafish
MeSH Terms
  • Animals
  • Blastula/cytology
  • Blastula/physiology*
  • Embryo, Nonmammalian/cytology
  • Embryo, Nonmammalian/physiology
  • Embryonic Development
  • Gene Expression Regulation, Developmental*
  • Morphogenesis
  • Zebrafish/embryology*
  • Zebrafish/physiology
  • Zebrafish Proteins/genetics
  • Zebrafish Proteins/metabolism
PubMed
30324595 Full text @ Meth. Mol. Biol.
Abstract
In the zebrafish embryo, cells of the early blastula animal pole are all equivalent and are fully pluripotent until the midblastula transition that occurs at the tenth cell cycle (512 to 1K cells). This naive territory of the embryo is therefore perfectly suited to assay for morphogen activity. Here we describe different methods to generate ectopic morphogen gradients, either in vivo at the animal pole of the embryo, or in vitro in animal pole explants or in aggregates of animal pole blastomeres (also named embryoid bodies). These methods include injection of mRNA coding for growth factor(s) into animal pole blastomere(s), transplantation of growth factor(s) secreting cells, implantation of beads coated with purified protein(s), and various combinations of these different approaches. Our comparative study reveals that all these methods allow to generate morphogen gradient(s) that are able to induce, both in vivo and in vitro, the formation of a well-patterned embryonic axis.
Genes / Markers
Figures
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Expression
Phenotype
Mutation and Transgenics
Human Disease / Model Data
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping
Errata and Notes