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Generation of clonal zebrafish line by androgenesis without egg irradiation

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Title: Generation of clonal zebrafish line by androgenesis without egg irradiation
Authors: Hou, Jilun Browse this author
Fujimoto, Takafumi Browse this author
Saito, Taiju Browse this author
Yamaha, Etsuro Browse this author →KAKEN DB
Arai, Katsutoshi Browse this author →KAKEN DB
Issue Date: 20-Aug-2015
Publisher: Nature Publishing Group
Journal Title: Scientific Reports
Volume: 5
Start Page: 13346
Publisher DOI: 10.1038/srep13346
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.
Type: article
Appears in Collections:水産科学院・水産科学研究院 (Graduate School of Fisheries Sciences / Faculty of Fisheries Sciences) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 荒井 克俊

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