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.
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
Figures
Show all Figures
Expression
Phenotype
Mutation and Transgenics
Human Disease / Model Data
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping
Errata and Notes