PUBLICATION
Ploidy Manipulation of Zebrafish Embryos with Heat Shock 2 Treatment
- Authors
- Baars, D.L., Takle, K.A., Heier, J., Pelegri, F.
- ID
- ZDB-PUB-170108-8
- Date
- 2016
- Source
- Journal of visualized experiments : JoVE (118): (Journal)
- Registered Authors
- Pelegri, Francisco
- Keywords
- none
- MeSH Terms
-
- Animals
- Diploidy
- Haploidy
- Heat-Shock Response/genetics*
- Hot Temperature
- Male
- Ploidies*
- Zebrafish/embryology*
- PubMed
- 28060351 Full text @ J. Vis. Exp.
Citation
Baars, D.L., Takle, K.A., Heier, J., Pelegri, F. (2016) Ploidy Manipulation of Zebrafish Embryos with Heat Shock 2 Treatment. Journal of visualized experiments : JoVE. (118).
Abstract
Manipulation of ploidy allows for useful transformations, such as diploids to tetraploids, or haploids to diploids. In the zebrafish Danio rerio, specifically the generation of homozygous gynogenetic diploids is useful in genetic analysis because it allows the direct production of homozygotes from a single heterozygous mother. This article describes a modified protocol for ploidy duplication based on a heat pulse during the first cell cycle, Heat Shock 2 (HS2). Through inhibition of centriole duplication, this method results in a precise cell division stall during the second cell cycle. The precise one-cycle division stall, coupled to unaffected DNA duplication, results in whole genome duplication. Protocols associated with this method include egg and sperm collection, UV treatment of sperm, in vitro fertilization and heat pulse to cause a one-cell cycle division delay and ploidy duplication. A modified version of this protocol could be applied to induce ploidy changes in other animal species.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping