Summary
During the last decade, zebrafish (Brachydanio rerio) have emerged as a novel and attractive system to study embryogenesis and organogenesis in vertebrates. The main reason is that both extensive genetic studies and detailed embryologic analysis are possible using this small tropical fresh water teleost. However, in vitro analysis using cell culture or molecular genetics are still far less advanced than in other vertebrate systems. Here we report the generation and characterization of a fibroblast like cell line, ZF4, derived from 1-day-old zebrafish embryos. The hyperploid cell line has been stable in multiple passages for more than 2 yr now and is the first zebrafish cell line that can be maintained in conventional medium containing mammalian serum. Using a series of plasmids for expression of a marker gene, we evaluate in ZF4 cells the relative strength of expression from several different viral, fish, and mammalian promoters. Stable integration can be obtained by using G418 selection. We hope that our cell line will be a useful tool for the analysis of gene regulation in zebrafish.
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Bayer, T. A.; Campos-Ortega, J. A. A transgene containing lacZ is expressed in primary sensory neurons in zebrafish. Development 115:421–426; 1992.
Bresch, H.; Beck, H.; Ehlermann, D., et al. A long-term toxicity test comprising reproduction and growth of zebrafish under 4-chloroaniline. Arch. Environ. Contam. Toxicol. 19:418–427; 1990.
Buono, R. J.; Linser, P. J. Transient expression of RSVCAT in transgenic zebrafish made by electroporation. Mol. Marine Biol. Biotechnol. 1:266–270; 1992.
Collodi, P.; Barnes, D. W. Mitogenic activity from trout embryos. Proc. Natl. Acad. Sci. USA 87:3498–3502; 1990.
Collodi, P.; Kamei, Y.; Ernst, T., et al. Culture of cells from zebrafish (Brachydanio rerio) embryo and adult tissue. Cell Biol. Toxicol. 8:43–61; 1992a.
Collodi, P.; Kamei, Y.; Sharps, A., et al. Fish embryo cell cultures for derivation of stem cells and transgenic chimeras. Mol. Marine Biol. Biotechnol. 1:257–265; 1992b.
Culp, P.; Nüsslein-Volhard, C.; Hopkins, N. High-frequency germ-line transmission of plasmid DNA sequences injected into fertilized zebrafish eggs. Proc. Natl. Acad. Sci. USA 88:7953–7957; 1991.
Edlund, T.; Walker, M. D.; Barr, P. J., et al. Cell specific expression of the rat insulin gene: evidence for role of two distinct 5′ flanking elements. Science 230:912–916; 1985.
Ekker, N. M.; Akimenko, M.-A.; Bremiller, R., et al. Regional expression of three homeobox transcripts in the inner ear of the zebrafish embryo. Neuron 9:27–35; 1992.
Endo, A.; Ingalls, T. Chromosomes of the zebrafish. J. Hered. 59:382–384; 1968.
Evans, M. H.; Kaufman, M. H. Establishment in culture of pluripotent cells from mouse embryos. Nature 292:154–156; 1981.
Friedenreich, H.; Schartl, M. Transient expression directed by homologous and heterologous promoter and enhancer sequences in fish cells. Nucleic Acids Res. 18:3299–3305; 1990.
Friedrich, G.; Soriano, P. Promoter traps in embryonic stem cells: a genetic screen to identify and mutate developmental genes in mice. Genes & Dev. 5:1513–1523; 1991.
Gatchalian, C. L.; Eisen, J. S. Pathway selection by ectopic motoneurons in embryonic zebrafish. Neuron 9:105–112; 1992.
Gillis, D. M.; Kramer, D. Ideal interference distributions: population density and patch use by zebrafish. Anim. Behav. 35:1875–1882; 1987.
Grunwald, D. J.; Streisinger, G. Induction of recessive lethal and specific locus mutations in the zebrafish with ethyl nitrosourea. Genet. Res. 59:103–116; 1992.
Gruss, P.; Khoury, G. Rescue of a splicing defective mutant by insertion of an heterologous intron. Nature 286:634–637; 1980.
Hatta, K.; Kimmel, C. B.; Ho, R. K., et al. The cyclops mutation blocks specification of the floor plate of the zebrafish central nervous system. Nature 350:339–341; 1991.
Hightower, L. E.; Renfro, J. L. Recent applications of fish cell culture in biomedical research. J. Exp. Zool. 248:290–302; 1988.
Ho, R. H.; Kane, D. Cell-autonomous action of zebrafishspt-1 mutation in specific mesodermal precursors. Nature 348:728–730; 1990.
Inoue, K. Expression of reporter genes introduced by microinjection and electroporation in fish embryos and fry. Mol. Marine Biol. Biotechnol. 1:266–270; 1992.
Kimmel, C. B.; Warga, R. M. Tissue specific cell lineages originate in the gastrula of the zebrafish. Science 231:365–368; 1986.
Kimmel, C. B.; Kane, D. A.; Walker, C., et al. A mutation that changes cell movement and cell fate in the zebrafish embryo. Nature 337:358–362; 1989.
Kirschbaum, F. Untersuchungen über das Farbmuster der ZebrabarbeBrachydanio rerio (Cyprinidae, Teleostei). Wilhelm Roux Arch. 177:129–152; 1975.
Krauss, S.; Maden, M.; Holder, N., et al. Zebrafish pax[b] is involved in the formation of the midbrain-hindbrain boundary. Nature 360:87–89; 1992.
Lin, S.; Long, W.; Chen, J., et al. Production of germ line chimeras in zebrafish by cell transplants from genetically pigmented to albino embryos. Proc. Natl. Acad. Sci. USA 89:4519–4523; 1992.
Lindl, T.; Bauer, J. Zell- and Gewebekultur. Stuttgart/New York: Gustav Fischer Verlag; 1989.
Liu, Z.; Moav, B.; Faras, A. J., et al. Development of expression vectors for transgenic fish. Bio/Technology 8:1268–1272; 1990.
Liu, Z.; Zhu, Z.; Roberg, K., et al. Isolation and characterization of theβ-actin gene of carp (Cyprinus carpio). DNA Sequence J. 1:125–136; 1990.
MacGregor, G. R.; Caskey, C. T. Construction of plasmids that expressE. coli β-galactosidase in mammalian cells. Nucleic Acids. Res. 17:2365; 1989.
Maniatis, T.; Fritsch, E. F.; Sambrook, J. Molecular cloning, a laboratory manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press; 1989.
Marcey, D.; Nüsslein-Volhard, C. Embryology goes fishing. Nature 321:380–381; 1986.
Miller, J. Experiments in molecular genetics. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press; 1972.
Moav, B.; Liu, Z.; Groll, Y., et al. Selection of promoters for gene transfer into fish. Mol. Marine Biol. Biotechnol. 1:338–345; 1992.
Molven, A.; Njolstad, P. R.; Fjose, A. Genomic structure and restricted neural expression of the zebrafishwnt-1 (int-1) gene. EMBO J. 10:799–807; 1991.
Ono, H.; Klein, D.; Vincek, V., et al. Major histocompatibility complex class II genes of zebrafish. Proc. Natl. Acad. Sci. USA 89:11886–11890; 1992.
Ozato, K.; Wakamatsu, Y.; Inoue, K. Medaka as a model of transgenic fish. Mol. Marine Biol. Biotechnol. 1:346–354; 1992.
Powers, D.; Hereford, L.; Cole, T., et al. Electroporation: a method for transferring genes into the gametes of zebrafish (Brachydanio rerio), channel catfish (Ictalurus punctatus) and common carp (Cyprinus carpio). Mol. Marine Biol. Biotechnol. 1:301–308; 1992.
Roosen-Runge, E. C. Furchung und Primitiventwicklung vonBrachydanio rerio. Anat. Anz. 81:297–301; 1936.
Schulte-Merker, S.; Ho, R. K.; Herrmann, B. G., et al. The protein product of the zebrafish homologue of the mouse T gene is expressed in nuclei of the germ ring and the notochord of the early embryo. Development 116:1021–1032; 1992.
St. Johnston, D.; Nüsslein-Volhard, C. The origin of pattern and polarity in theDrosophila embryo. Cell 68:201–219; 1992.
Streisinger, G. F.; Walker, C.; Dower, D., et al. Production of clones of homozygous diploid zebrafish (Brachydanio rerio). Nature 291:293–296; 1981.
Stuart, G. W.; McMurray, J. V.; Westerfield, M. Replication, integration, and stable germ line transmission of foreign sequences injected into early zebrafish embryos. Development 103:403–412; 1988.
Stuart, G. W.; Vielkind, J. R.; McMurray, J. V., et al. Stable lines of transgenic zebrafish exhibit reproducible patterns of transgenic expression. Development 109:577–584; 1990.
Westerfield, M. The zebrafish book. Eugene: University of Oregon Press; 1989.
Wilkins, A. S. Genetic analysis of animal development. New York: Wiley & Sons; 1993.
Winkler, C.; Hong, Y.; Wittbrodt, J., et al. Analysis of heterologous and homologous promoters and enhancers in vitro and in vivo by gene transfer into Japanese Medaka (Oryzias latipes) andXiphophorus. Mol. Marine Biol. Biotechnol. 1:326–337; 1992.
Wolf, K.; Quimby, M. C. Established eurythermic line of fish cells in vitro. Science 137:1065–1066; 1962.
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Driever, W., Rangini, Z. Characterization of a cell line derived from zebrafish (brachydanio rerio) embryos. In Vitro Cell Dev Biol - Animal 29, 749–754 (1993). https://doi.org/10.1007/BF02631432
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DOI: https://doi.org/10.1007/BF02631432