Abstract
Metazoan animals are typically diploid, possessing two sets of a chromosome in the somatic cells of an organism. In naturally diploid species, alteration from the endogenous diploid state is usually embryonic lethal. However, the ability to experimentally manipulate ploidy of animal embryos has fundamental as well as applied biology advantages. In this chapter we describe experimental procedures to convert normally diploid zebrafish embryos into haploid or tetraploid states. We also describe methodologies to verify the ploidy of embryos and the utility of ploidy manipulation in expediting the isolation of mutations using both forward and reverse genetic strategies in zebrafish.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Endersby J (2007) A Guinea Pig’s History of biology: the plants and animals who taught us the facts of life. Harvard University Press, Cambridge, MA
Schön I, Martens K, van Dijk PJ (2009) Lost sex: the evolutionary biology of parthenogenesis. Springer, Dordrecht
Kubiak J, Paldi A, Weber M, Maro B (1991) Genetically identical parthenogenetic mouse embryos produced by inhibition of the first meiotic cleavage with cytochalasin D. Development 111:763–769
Leeb M, Wutz A (2011) Derivation of haploid embryonic stem cells from mouse embryos. Nature 479:131–134
Li W, Shuai L, Wan H, Dong M, Wang M, Sang L et al (2012) Androgenetic haploid embryonic stem cells produce live transgenic mice. Nature 490:407–411
Komma DJ, Endow SA (1995) Haploidy and androgenesis in Drosophila. Proc Natl Acad Sci U S A 92:11884–11888
Araki K, Okamoto H, Graveson AC, Nakayama I, Nagoya H (2001) Analysis of haploid development based on expression patterns of developmental genes in the medaka Oryzias latipes. Develop Growth Differ 43:591–599
Corley-Smith GE, Lim CJ, Brandhorst BP (1996) Production of androgenetic zebrafish (Danio rerio). Genetics 142:1265–1276
Gurdon JB (1960) The effects of ultraviolet radiation on uncleaved eggs of Xenopus laevis. J Cell Sci S3:299–311
Streisinger G, Walker C, Dower N, Knauber D, Singer F (1981) Production of clones of homozygous diploid zebra fish (Brachydanio rerio). Nature 291:293–296
Moens CB, Yan YL, Appel B, Force AG, Kimmel CB (1996) Valentino: a zebrafish gene required for normal hindbrain segmentation. Development 122:3981–3990
Walker C (1999) Haploid screens and gamma-ray mutagenesis. Methods Cell Biol 60:43–70
Yi M, Hong N, Hong Y (2009) Generation of medaka fish haploid embryonic stem cells. Science 326:430–433
Heier J, Takle KA, Hasley AO, Pelegri F (2015) Ploidy manipulation and induction of alternate cleavage patterns through inhibition of centrosome duplication in the early zebrafish embryo. Dev Dyn 244:1300–1312
Amsterdam A, Burgess S, Golling G, Chen W, Sun Z, Townsend K et al (1999) A large-scale insertional mutagenesis screen in zebrafish. Genes Dev 13:2713–2724
Driever W, Solnica-Krezel L, Schier AF, Neuhauss SC, Malicki J, Stemple DL et al (1996) A genetic screen for mutations affecting embryogenesis in zebrafish. Development 123:37–46
Haffter P, Granato M, Brand M, Mullins MC, Hammerschmidt M, Kane DA (1996) The identification of genes with unique and essential functions in the development of the zebrafish, Danio rerio. Development 123:1–36
Patton EE, Zon LI (2001) The art and design of genetic screens: zebrafish. Nat Rev Genet 2:956–966
van Eeden FJM, Granato M, Odenthal J, Haffter P (1998) Chapter 2: Developmental mutant screens in the zebrafish. Methods Cell Biol 60:21–41
Pelegri F, Dekens MP, Schulte-Merker S, Maischein HM, Weiler C et al (2004) Identification of recessive maternal-effect mutations in the zebrafish using a gynogenesis-based method. Dev Dyn 231:324–335
Pelegri F, Mullins MC (2004) Genetic screens for maternal-effect mutations. Methods Cell Biol 77:21–51
Pelegri F, Mullins MC (2016) Genetic screens for mutations affecting adult traits and parental-effect genes. Methods Cell Biol 135:39–87
Pelegri F, Schulte-Merker S (1999) Chapter 1: A gynogenesis-based screen for maternal-effect genes in the zebrafish Danio rerio. Methods Cell Biol 60:1–20
Menon T, Nair S (2018) A transient window of resilience during early development minimizes teratogenic effects of heat in zebrafish embryos. Dev Dyn 247(8):992–1004
Acknowledgments
Our research is funded by a Wellcome Trust/Department of Biotechnology India Alliance Intermediate Fellowship (IA/I/13/2/501042) to SN and by the Tata Institute of Fundamental Research, Mumbai.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Menon, T., Nair, S. (2019). Experimental Manipulation of Ploidy in Zebrafish Embryos and Its Application in Genetic Screens. In: Pelegri, F. (eds) Vertebrate Embryogenesis. Methods in Molecular Biology, vol 1920. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9009-2_8
Download citation
DOI: https://doi.org/10.1007/978-1-4939-9009-2_8
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-9008-5
Online ISBN: 978-1-4939-9009-2
eBook Packages: Springer Protocols