PUBLICATION
Making gynogenetic diploid zebrafish by early pressure
- Authors
- Walker, C., Walsh, G.S., and Moens, C.
- ID
- ZDB-PUB-090706-17
- Date
- 2009
- Source
- Journal of visualized experiments : JoVE (28): (Journal)
- Registered Authors
- Moens, Cecilia, Walker, Charline, Walsh, Gregory
- Keywords
- none
- MeSH Terms
-
- Animals
- Diploidy*
- Female
- Genetic Techniques
- Homozygote
- Male
- Ovum/physiology
- Spermatozoa/physiology
- Spermatozoa/radiation effects
- Ultraviolet Rays
- Zebrafish/genetics*
- PubMed
- 19568220 Full text @ J. Vis. Exp.
Citation
Walker, C., Walsh, G.S., and Moens, C. (2009) Making gynogenetic diploid zebrafish by early pressure. Journal of visualized experiments : JoVE. (28).
Abstract
Heterozygosity in diploid eukaryotes often makes genetic studies cumbersome. Methods that produce viable homozygous diploid offspring directly from heterozygous females allow F1 mutagenized females to be screened directly for deleterious mutations in an accelerated forward genetic screen. Streisinger et al. described methods for making gynogenetic (homozygous) diploid zebrafish by activating zebrafish eggs with ultraviolet light-inactivated sperm and preventing either the second meiotic or the first zygotic cell division using physical treatments (heat or pressure) that deploymerize microtubules. The "early pressure" (EP) method blocks the meiosis II, which occurs shortly after fertilization. The EP method produces a high percentage of viable embryos that can develop to fertile adults of either sex. The method generates embryos that are homozygous at all loci except those that were separated from their centromere by recombination during meiosis I. Homozygous mutations are detected in EP clutches at between 50% for centromeric loci and less than 1% for telomeric loci. This method is reproduced verbatim from the Zebrafish Book.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping