PUBLICATION

Generation of clonal zebrafish line by androgenesis without egg irradiation

Authors
Hou, J., Fujimoto, T., Saito, T., Yamaha, E., Arai, K.
ID
ZDB-PUB-150822-15
Date
2015
Source
Scientific Reports   5: 13346 (Journal)
Registered Authors
Keywords
none
MeSH Terms
  • Fertilization
  • Ovum/radiation effects*
  • Clone Cells
  • Heat-Shock Response
  • Cell Line
  • Female
  • Haploidy
  • Chromosomes/genetics
  • Male
  • Zebrafish/metabolism*
  • Animals
  • Cold Temperature
(all 12)
PubMed
26289165 Full text @ Sci. Rep.
Abstract
Generation of clonal zebrafish will facilitate large-scale genetic screening and help us to overcome other biological and biotechnological challenges due to their isogenecity. However, protocols for the development of clonal lines have not been optimized. Here, we sought to develop a novel method for generation of clonal zebrafish by androgenesis induced by cold shock. Androgenetic zebrafish doubled haploids (DHs) were induced by cold shock of just-fertilized eggs, and the eggs were then heat shocked to double the chromosome set. The yield rate of putative DHs relative to the total number of eggs used was 1.10% ± 0.19%. Microsatellite genotyping of the putative DHs using 30 loci that covered all 25 linkage groups detected no heterozygous loci, confirming the homozygosity of the DHs. Thus, a clonal line was established from sperm of a DH through a second cycle of cold-shock androgenesis and heat-shock chromosome doubling, followed by genetic verification of the isogenic rate confirming the presence of identical DNA fingerprints by using amplified fragment length polymorphism markers. In addition, our data provided important insights into the cytological mechanisms of cold-shock-induced androgenesis.
Genes / Markers
Figures
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Phenotype
No data available
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
slc24a5_unspecified
    		Unspecified
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    Human Disease / Model
    Sequence Targeting Reagents
    Fish
    Antibodies
    Orthology
    Engineered Foreign Genes
    No data available
    Mapping
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    Citation
    Hou, J., Fujimoto, T., Saito, T., Yamaha, E., Arai, K. (2015) Generation of clonal zebrafish line by androgenesis without egg irradiation. Scientific Reports. 5:13346.