ZFIN ID: ZDB-PUB-210220-27
Masculinization of Zebrafish Through Partial Depletion of Primordial Germ Cells by Injecting Diluted Morpholino Oligonucleotides into Embryos
Orbán, L., Saju, J.M., Tzung, K.W., Liew, W.C.
Date: 2021
Source: Methods in molecular biology (Clifton, N.J.)   2218: 49-60 (Chapter)
Registered Authors: Liew, Woei Chang, Saju, Jolly M.
Keywords: Gonadal transcriptome, Isolation of early gonads, Sex bias, Testis development,  Primordial germ cells
MeSH Terms:
  • Animals
  • Embryo, Nonmammalian/drug effects*
  • Embryo, Nonmammalian/metabolism
  • Female
  • Germ Cells/drug effects*
  • Germ Cells/metabolism
  • Gonads/drug effects
  • Gonads/metabolism
  • Injections
  • Male
  • Morpholinos/administration & dosage*
  • Oligonucleotides/administration & dosage*
  • RNA-Binding Proteins/metabolism
  • Zebrafish/metabolism
  • Zebrafish Proteins/metabolism
PubMed: 33606222 Full text @ Meth. Mol. Biol.
ABSTRACT
The regulation of reproduction in zebrafish, the prime model of fish research, is not fully understood. An efficient tool to gain a better understanding of this complicated process is utilization of severely sex-biased families or groups. Here, we describe a method for partial depletion of primordial germ cells (PGCs) that leads to eventual masculinization of zebrafish. The technique is based on injecting early embryos with diluted morpholino oligonucleotides that temporarily interfere with the production of Dead end (Dnd), an RNA-binding protein essential for PGC survival. In addition, we also propose the use of eviscerated trunk, as a suitable alternative for examining gonadal expression in juvenile zebrafish.
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