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Germline Development in the Zebrafish pp 49–60Cite as

Masculinization of Zebrafish Through Partial Depletion of Primordial Germ Cells by Injecting Diluted Morpholino Oligonucleotides into Embryos

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 2218))

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.

The methodologies described in this chapter were developed at Temasek Life Sciences Laboratory (TLL), Singapore, during the period between 2006 and 2017. All four authors were affiliated with the Reproductive Genomics Group of TLL during the above time period. L.O. and K.W.T. are no longer with TLL, whereas J.M.S. and L.W.C. have moved to different research groups within TLL.

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Acknowledgments

The work described in the MS was funded by the National Research Foundation, Prime Minister’s Office, Singapore, under its Competitive Research Programme (Award No: NRF-CRP7-2010-001) and by internal research grants from Temasek Life Sciences Laboratory. During the writing of this review, L.O. was supported by the Frontline Research Grant of the National Research, Development and Innovation Office of Hungary (KKP126764). We would like to thank Lisbeth Olsen for the transgenic line and Lydia Tan for helping with the figures.

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Author notes

  1. László Orbán

    Present address: Frontline Fish Genomics Research Group, Department of Animal Sciences, Georgikon Campus, Szent István University, Keszthely, Hungary

  2. Keh-Weei Tzung

    Present address: Institute of Molecular and Cell Biology (IMCB), A*STAR, Singapore, Singapore

Authors and Affiliations

  1. Reproductive Genomics Group, Temasek Life Sciences Laboratory, Singapore, Singapore

    László Orbán, Jolly M. Saju, Keh-Weei Tzung & Woei Chang Liew

  2. Centre for Comparative Genomics, Murdoch University, Murdoch, WA, Australia

    László Orbán

Authors
  1. László Orbán
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  2. Jolly M. Saju
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  3. Keh-Weei Tzung
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  4. Woei Chang Liew
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Corresponding author

Correspondence to László Orbán .

Editor information

Editors and Affiliations

  1. Institute for Developmental Biochemistry, University of Göttingen Medical Center Georg-August-Universität, Göttingen, Germany

    Roland Dosch

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Orbán, L., Saju, J.M., Tzung, KW., Liew, W.C. (2021). Masculinization of Zebrafish Through Partial Depletion of Primordial Germ Cells by Injecting Diluted Morpholino Oligonucleotides into Embryos. In: Dosch, R. (eds) Germline Development in the Zebrafish. Methods in Molecular Biology, vol 2218. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0970-5_5

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  • DOI: https://doi.org/10.1007/978-1-0716-0970-5_5

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0969-9

  • Online ISBN: 978-1-0716-0970-5

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