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Induction of recessive lethal and specific locus mutations in the zebrafish with ethyl nitrosourea

Published online by Cambridge University Press:  14 April 2009

David Jonah Grunwald
Affiliation:
Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403
George Streisinger
Affiliation:
Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403
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Summary

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Recessive lethal mutations and mutations at the gol-1 locus were induced in the zebrafish by exposure of mature sperm to the alkylating agent ethyl nitrosourea (ENU). Embryonic lethal phenotypes were recognized among the parthenogenetic progeny of mutagenized animals or among the progeny of daughters of mutagenized animals. Novel specific locus mutations were identified by the failure of mutagenized chromosomes to complement pre-existing mutant alleles at the gol-1 locus. Each mutagenized individual harboured approximately 10 embryonic lethal mutations in its germ line and about 1 in 500 mutagenized animals harboured a new mutation at the gol-1 locus. Three lines of evidence indicate that the majority of mutations that were recovered following treatment of mature sperm with ENU were probably point mutations. First, the soma and germ lines of mutagenized animals were mosaic, as expected following simple alkylation of sperm DNA. Second, mutations induced by ENU at the gol-1 locus affected pigmentation but not viability, unlike the majority of mutations induced at this locus with y-irradiation. Third, the ratio of specific locus: recessive lethal mutations induced by ENU was approximately 50-fold lower than the ratio observed following mutagenesis with y-rays. Comparison of the incidence with which embryonic recessive lethal mutations were induced with the incidence with which specific locus mutations arose indicates that there are greater than 5000 genes essential to the development and viability of the zebrafish embryo.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1992

References

Ames, B. N., McCann, J. & Yamasaki, E. (1975). Methods for detecting carcinogens and mutagens with the Salmonella/mammalian-microsome mutagenicity test. Mutation Research 31, 347363.CrossRefGoogle ScholarPubMed
Bode, V. C. (1984). Ethylnitrosourea mutagenesis and the isolation of mutant alleles for specific genes located in the T region of mouse chromosome 17. Genetics 108,457470.CrossRefGoogle Scholar
Brenner, S. (1974). The genetics of Caenorhabditis elegans. Genetics 77, 7194.CrossRefGoogle ScholarPubMed
Chakrabarti, S., Streisinger, G., Singer, F. & Walker, C. (1983). Frequency of y-ray induced specific locus and recessive lethal mutations in mature germ cells of the zebrafish, Brachydanio rerio. Genetics 103, 109123.CrossRefGoogle Scholar
Clifford, R. J. & Schupbach, T. (1989). Coordinately and differentially mutable activities of torpedo, theDrosophila melanogaster homolog of the vertebrate EGF receptor gene. Genetics 123, 771787.CrossRefGoogle Scholar
Eberl, D. F. & Hilliker, A. J. (1988). Characterization of Xlinked recessive lethal mutations affecting embryonic morphogenesis inDrosophila melanogaster. Genetics 118, 109120.CrossRefGoogle Scholar
Grunwald, D. J., Kimmel, C. B, Westerfield, M., Walker, C. & Streisinger, G. (1988). A neural degeneration mutation that spares primary neurons in the zebrafish. Cellularity Biology 126, 115128.Google ScholarPubMed
Grunwald, D. J. & Streisinger, G. (1991). Induction of mutations in the zebrafish with ultraviolet light. Genetical Research 59, 93101.CrossRefGoogle Scholar
Hince, T. A. & Neale, S. (1974). A comparison of the mutagenic action of the methyl and ethyl derivatives of nitrosamides and nitrosamidines on Escherichia coli. Mutation Research 24, 383387.CrossRefGoogle ScholarPubMed
Johnson, F. M. & Lewis, S. E. (1981). Electrophoretically detected germinal mutations induced in the mouse by ethyinitrosourea. Proceedings of the National Academy of Sciences, USA 78, 31383141.CrossRefGoogle Scholar
Jurgens, G., Wieschaus, E., Nussleinn-Volhard, C. & Kluding, H. (1984). Mutations affecting the pattern of the larval cuticle inDrosophila melanogaster. II. Zygotic loci on the third chromosome. Roux's Archives of Cellularity Biology 193, 283295.Google ScholarPubMed
Kimmel, C. B., Kane, D. A., Walker, C, Warga, R. M. & Rothman, M. B. (1989). A mutation that changes cell movement and cell fate in the zebrafish embryo. Nature 337, 358362.CrossRefGoogle ScholarPubMed
King, T. R., Dove, W. F., Herrmann, B., Moser, A. R. & Shedlovsky, A. (1989). Mapping to molecular resolution in the T to H-2 region of the mouse genome with a nested set of meiotic recombinants. Proceedings of the National Academy of Sciences, USA 86, 222226.CrossRefGoogle Scholar
Lee, K., Gold, B. & Mirvish, S. S. (1977). Mutagenicity of 22 N-nitrosamides and related compounds for Salmonella typhimurium TA1535. Mutation Research 48, 131138.CrossRefGoogle ScholarPubMed
Loveless, A. & Hampton, C. L. (1969). Inactivation and mutation of coliphage T2 by N-methyl and N-ethyl-N-nitrosourea. Mutation Research 7, 112.CrossRefGoogle ScholarPubMed
Moser, A. R., Pitot, H. C. & Dove, W. F. (1990). A dominant mutation that predisposes to multiple intestinal neoplasia in the mouse. Science 247, 322324.CrossRefGoogle ScholarPubMed
Muller, H. J. & Altenberg, E. (1919). The rate of change of hereditary factors in Drosophila. Proceedings of the Society of Experimental Biology and Medicine 17, 1014.CrossRefGoogle Scholar
Nusslein-Volhard, C, Wieschaus, E. & Kluding, H. (1984). Mutations affecting the pattern of the larval cuticle inDrosophila melanogaster. I. Zygotic loci on the second chromosome. Roux's Archives of Cellularity Biology 193, 267282.Google ScholarPubMed
Pastnik, A., Vreeken, C, Nivard, M. J. M., Searles, L. L. & Vogel, E. W. (1989). Sequence analysis of A^-ethyl-A^-nitrosourea-induced vermilion mutations inDrosophila melanogaster. Genetics 123, 123129.CrossRefGoogle Scholar
Richardson, K. K., Richardson, F. C, Crosby, R. M., Swenberg, J. A. & Skopek, T. R. (1987). DNA base changes and alkylation following in vivo exposure of Escherichia coli to N-methyl-N-nitrosourea or N-ethyl-N-nitrosourea. Proceedings of the National Academy of Sciences, USA 84, 344348.CrossRefGoogle ScholarPubMed
Russell, W. L., Kelly, E. M., Hunsicker, P. R., Bangham, J. W., Maddux, S. C. & Phipps, E. L. (1979). Specificlocus test shows ethylnitrosourea to be the most potent mutagen in the mouse. Proceedings of the National Academy of Sciences, USA 76, 58185819.CrossRefGoogle ScholarPubMed
Shedlovsky, A., King, T. R. & Dove, W. F. (1988). Saturation germ line mutagenesis of the murine / region including a lethal allele at the quaking locus. Proceedings of the National Academy of Sciences, USA 85, 180184.CrossRefGoogle ScholarPubMed
Streisinger, G., Walker, C, Dower, N., Knauber, D. & Singer, F. (1981). Production of clones of homozygous diploid zebrafish (Brachydanio rerio). Nature (London) 291, 293296.CrossRefGoogle Scholar
Streisinger, G., Singer, F., Walker, C, Knauber, D. & Dower, N. (1986). Segregation analyses and genecentromere distances in zebrafish. Genetics 112, 311319.CrossRefGoogle ScholarPubMed
Streisinger, G., Coale, F., Taggart, C, Walker, C. & Grunwald, D. J. (1989). Clonal origins of cells in the pigmented retina of the zebrafish eye. Cellularity Biology 131, 6069.Google ScholarPubMed
Walker, C. & Streisinger, G. (1983). Induction of mutations by y-rays in pregonial germ cells of zebrafish embryos. Genetics 103, 125136.CrossRefGoogle Scholar
Wieschaus, E., Nusslein-Volhard, C. & Jurgens, G. (1984). Mutations affecting the pattern of the larval cuticle inDrosophila melanogaster. III. Zygotic loci on the Xchromosome and fourth chromosome. Roux's Archives of Cellularity Biology 193, 296307.Google ScholarPubMed
Wood, W. B. (1988). The Nematode Caenorhabditis elegans. Cold Spring Harbour, New York: Cold Spring Harbour Laboratory.Google Scholar