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
-
- Gene Expression Regulation, Developmental/genetics
- Gene Expression Regulation, Developmental/physiology
- Zebrafish Proteins/genetics
- Morpholinos/metabolism
- Transcription, Genetic/genetics
- Transcription, Genetic/physiology*
- Zygote/metabolism
- Zygote/physiology
- Female
- Genome/genetics
- Genome/physiology
- Zebrafish/genetics
- Zebrafish/metabolism
- Zebrafish/physiology*
- Male
- Animals
- Embryonic Development/genetics
- Embryonic Development/physiology
- Embryo, Nonmammalian/metabolism
- Embryo, Nonmammalian/physiology
- Germ Cells/metabolism
- Germ Cells/physiology*
- PubMed
- 33606232 Full text @ Meth. Mol. Biol.
Citation
D'Orazio, F.M., Jasiulewicz, A., Hadzhiev, Y., Müller, F. (2021) Visualization of Transcriptional Activity in Early Zebrafish Primordial Germ Cells. Methods in molecular biology (Clifton, N.J.). 2218:185-194.
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
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping