PUBLICATION

Visualization of Transcriptional Activity in Early Zebrafish Primordial Germ Cells

Authors
D'Orazio, F.M., Jasiulewicz, A., Hadzhiev, Y., Müller, F.
ID
ZDB-PUB-210220-36
Date
2021
Source
Methods in molecular biology (Clifton, N.J.)   2218: 185-194 (Chapter)
Registered Authors
Hadzhiev, Yavor, Müller, Ferenc
Keywords
Bucky ball, Germ cells, Germ plasm, Microinjections, Microscopy, Morpholinos, Transcription, ZGA, Zebrafish
MeSH Terms
  • Animals
  • Embryo, Nonmammalian/metabolism
  • Embryo, Nonmammalian/physiology
  • Embryonic Development/genetics
  • Embryonic Development/physiology
  • Female
  • Gene Expression Regulation, Developmental/genetics
  • Gene Expression Regulation, Developmental/physiology
  • Genome/genetics
  • Genome/physiology
  • Germ Cells/metabolism
  • Germ Cells/physiology*
  • Male
  • Morpholinos/metabolism
  • Transcription, Genetic/genetics
  • Transcription, Genetic/physiology*
  • Zebrafish/genetics
  • Zebrafish/metabolism
  • Zebrafish/physiology*
  • Zebrafish Proteins/genetics
  • Zygote/metabolism
  • Zygote/physiology
PubMed
33606232 Full text @ Meth. Mol. Biol.
Abstract
Here, we describe a fast and straightforward methodology to in vivo detect transcriptional activity in the early zebrafish germ line. We report how fluorescently labeled morpholinos, targeted to nascent early transcripts, can be used to track the onset of transcriptional events during early embryogenesis. This method could be applied to any tagged cell line in a developing early zebrafish embryo as long as the gene of interest is expressed at high enough level for morpholino detection and is expressed at the first and main wave of genome activation, for which the protocol has been verified. The protocol, in combination with genetic manipulation, allows studies of mechanisms driving zygotic genome activation (ZGA) in individual cells. The reported procedures apply to a broad range of purposes for zebrafish embryo manipulation in view of imaging nuclear molecules in specific cell types.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping