Visualization of Transcriptional Activity in Early Zebrafish Primordial Germ Cells

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 2218; p. 185
• Main Authors: D'Orazio, Fabio M, Jasiulewicz, Aleksandra, Hadzhiev, Yavor, Müller, Ferenc
• Format: Journal Article
• Language: English
• Published: United States 2021
• Subjects: 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; Bucky ball; Germ cells; Microinjections; Microscopy; Transcription; ZGA; Zebrafish; Morpholinos; Germ plasm
• ISSN: 1940-6029
• DOI: 10.1007/978-1-0716-0970-5_15

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.