PUBLICATION

Large-scale mutagenesis in the zebrafish: in search of genes controlling development in a vertebrate

Authors
Mullins, M.C., Hammerschmidt, M., Haffter, P., and Nüsslein-Volhard, C.
ID
ZDB-PUB-961014-809
Date
1994
Source
Current biology : CB   4: 189-202 (Journal)
Registered Authors
Haffter, Pascal, Hammerschmidt, Matthias, Mullins, Mary C., Nüsslein-Volhard, Christiane
Keywords
none
MeSH Terms
  • Animal Husbandry
  • Animals
  • Crosses, Genetic
  • Embryonic and Fetal Development/genetics
  • Ethyl Methanesulfonate
  • Ethylnitrosourea
  • Female
  • Gene Deletion
  • Genes, Lethal
  • Male
  • Meiosis/genetics
  • Mitosis/genetics
  • Mutagenesis
  • Pilot Projects
  • Point Mutation
  • Spermatogenesis/genetics
  • Zebrafish/embryology
  • Zebrafish/genetics*
PubMed
7922324 Full text @ Curr. Biol.
Abstract
BACKGROUND: In Drosophila melanogaster and Caenorhabditis elegans, the elucidation of developmental mechanisms has relied primarily on the systematic induction and isolation of mutations in genes with specific functions in development. Such an approach has not yet been possible in a vertebrate species, owing to the difficulty of analyzing and keeping a sufficiently high number of mutagenized lines of animals. RESULTS: We have developed the methods necessary to perform large-scale saturation screens for mutations affecting embryogenesis in the zebrafish, Danio (Brachydanio) rerio. Firstly, a new aquarium system was developed to raise and keep large numbers of strains of genetically different fish safely and with little maintenance care. Secondly, by placing adult male fish in water containing the chemical mutagen, ethylnitrosourea, we induced point mutations in premeiotic germ cells with a rate of one to three mutations per locus per 1,000 mutagenized haploid genomes. This rate, which is similar to the mutagenesis rates produced by ethylmethanesulfonate in Drosophila, was determined for alleles at four different pigmentation genes. Finally, in a pilot screen in which mutagenized fish were inbred for two generations and scored for embryonic mutants, we isolated 100 recessive mutations with phenotypes visible in the homozygous embryos. CONCLUSION: The high rate of induction and recovery of point mutations, in addition to an efficient aquarium system to house large numbers of mutagenized lines, means that it is now possible to perform saturation mutagenesis screens in a vertebrate, similar to those done in invertebrates.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping