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
-
- Animals
- Cell Line
- Chromosomes/genetics
- Clone Cells
- Cold Temperature
- Female
- Fertilization
- Haploidy
- Heat-Shock Response
- Male
- Ovum/radiation effects*
- Zebrafish/metabolism*
- PubMed
- 26289165 Full text @ Sci. Rep.
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.
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
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping