Visualization of transcriptional activity in early zebrafish primordial germ cells

Fabio M. D’Orazio; Aleksandra Jasiulewicz; Yavor Hadzhiev; Ferenc Müller

doi:10.1007/978-1-0716-0970-5_15

Visualization of transcriptional activity in early zebrafish primordial germ cells

Fabio M. D’Orazio, Aleksandra Jasiulewicz, Yavor Hadzhiev, Ferenc Müller^*

^*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceeding › Chapter

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.

Original language	English
Title of host publication	Germline Development in the Zebrafish
Subtitle of host publication	Methods and Protocols
Editors	Roland Dosch
Publisher	Humana Press
Pages	185-194
Number of pages	10
Edition	1
ISBN (Electronic)	9781071609705
ISBN (Print)	9781071609699, 9781071609729
DOIs	https://doi.org/10.1007/978-1-0716-0970-5_15
Publication status	Published - 20 Feb 2021

Publication series

Name	Methods in Molecular Biology
Volume	2218
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Bibliographical note

Funding Information:
This work was supported by the Horizon 2020 programme Zencode-ITN, Wellcome Trust Investigator award (106955/Z/ 15/Z) to F.M., Horizon 2020 FET project to F.M. We thank Roland Dosch at the University Medical Center in G?ttingen for providing the Tg(Buc-GFP) line and the BMSU facility at the University of Birmingham for the maintenance.

Publisher Copyright:
© Springer Science+Business Media, LLC, part of Springer Nature 2021.

Keywords

Bucky ball
Germ cells
Germ plasm
Microinjections
Microscopy
Morpholinos
Transcription
Zebrafish
ZGA

ASJC Scopus subject areas

Molecular Biology
Genetics

Access to Document

10.1007/978-1-0716-0970-5_15

Cite this

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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.",

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note = "Funding Information: This work was supported by the Horizon 2020 programme Zencode-ITN, Wellcome Trust Investigator award (106955/Z/ 15/Z) to F.M., Horizon 2020 FET project to F.M. We thank Roland Dosch at the University Medical Center in G?ttingen for providing the Tg(Buc-GFP) line and the BMSU facility at the University of Birmingham for the maintenance. Publisher Copyright: {\textcopyright} Springer Science+Business Media, LLC, part of Springer Nature 2021.",

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Visualization of transcriptional activity in early zebrafish primordial germ cells. / D’Orazio, Fabio M.; Jasiulewicz, Aleksandra; Hadzhiev, Yavor et al.
Germline Development in the Zebrafish: Methods and Protocols. ed. / Roland Dosch. 1. ed. Humana Press, 2021. p. 185-194 (Methods in Molecular Biology; Vol. 2218).

Research output: Chapter in Book/Report/Conference proceeding › Chapter

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Visualization of transcriptional activity in early zebrafish primordial germ cells

Abstract

Publication series

Bibliographical note

Keywords

ASJC Scopus subject areas

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