Summary
It is largely accepted that vertebrates are more susceptible to chemical insult during the early life stage. It is implied that if a chemical such as ethanol is developmentally toxic, it must interfere with, or modulate, critical signaling pathways. The probable molecular explanation for increased embryonic susceptibility is that collectively there is no other period of an animal's lifespan when the full repertoire of molecular signaling is active. Understanding the mechanism by which ethanol exposure disrupts vertebrate embryonic development is enormously challenging; it requires a thorough understanding of the normal molecular program to understand how transient ethanol exposure disrupts signaling and results in detrimental long-lasting effects. During the past several years, investigators have recognized the advantages of the zebrafish model to discover the signaling events that choreograph embryonic development. External development coupled with the numerous molecular and genetic methods make this model a valuable tool to unravel the mechanisms by which ethanol disrupts embryonic development. In this chapter we describe procedures used to evaluate and define the morphological, cellular and molecular responses to ethanol in zebrafish.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
1. Becker, H. C., Diaz-Granados, J. L., and Randall, C. L. (1996) Teratogenic actions of ethanol in the mouse: a minireview. Pharmacol. Biochem. Behav. 55, 501–513.
2. Brien, J. F., Clarke, D. W., Richardson, B., and Patrick, J. (1985) Disposition of ethanol in maternal blood, fetal blood, and amniotic fluid of third-trimester pregnant ewes. Am. J. Obstet. Gynecol. 152, 583–590.
3. Richardson, B. S., Patrick, J. E., Bousquet, J., Homan, J., and Brien, J. F. (1985) Cerebral metabolism in fetal lamb after maternal infusion of ethanol. Am. J. Physiol. 249, R505–R509.
4. Bupp Becker, S. R., and Shibley, I. A., Jr. (1998) Teratogenicicty of ethanol in different chicken strains. Alcohol 33, 457–464.
5. Su, B., Debelak, K. A., Tessmer, L. L., Cartwright, M. M., and Smith, S. M. (2001) Genetic influences on craniofacial outcome in an avian model of prenatal alcohol exposure. Alcohol Clin. Exp. Res. 25, 60–69.
6. Ranganathan, S., Davis, D. G., and Hood, R. D. (1987) Developmental toxicity of ethanol in Drosophila melanogaster. Teratology 36, 45–49.
7. Ranganathan, S., Davis, D. G., Leeper, J. D., and Hood, R. D. (1987) Effects of differential alcohol dehydrogenase activity on the developmental toxicity of ethanol in Drosophila melanogaster. Teratology 36, 329–334.
8. Dhawan, R., Dusenbery, D. B., and Williams, P. L. (1999) Comparison of lethality, reproduction, and behavior as toxicological endpoints in the nematode Caenorhabditis elegans. J. Toxicol. Environ. Health A 58, 451–462.
9. Thompson, G. and de Pomerai, D. I. (2005) Toxicity of short-chain alcohols to the nematode Caenorhabditis elegans: A comparison of endpoints. J. Biochem. Mol. Toxicol. 19, 87–95.
10. Laale, H. W. (1971) Ethanol induced notochord and spinal cord duplications in the embryo of the zebrafish, Brachydanio rerio. J. Exp. Zool. 177, 51–64.
11. Reimers, M. J., Flockton, A. R., and Tanguay, R. L. (2004) Ethanol and acetaldehyde-mediated developmental toxicity in zebrafish. Neurotoxicol. Teratol. 26, 769–781.
12. Ackermann, G. E., and Paw, B. H. (2003) Zebrafish: a genetic model for vertebrate organogenesis and human disorders. Front. Biosci. 8, d1227–d1253.
13. Rubinstein, A. L. (2003) Zebrafish: from disease modeling to drug discovery. Curr. Opin. Drug Discov. Devel. 6, 218–23.
14. Rodriguez, F., Lopez, J. C., Vargas, J. P., Broglio, C., Gomez, Y., and Salas, C. (2002) Spatial memory and hippocampal pallium through vertebrate evolution: Insights from reptiles and teleost fish. Brain Res. Bull. 57, 499–503.
15. Amsterdam, A., S. Lin, L.G. Moss, and N. Hopkins (1996) Requirements for green fluorescent protein detection in transgenic zebrafish embryos. Gene 173, 99–103.
16. Higashijima, S., Okamoto, H., Ueno, N., Hotta, Y., and Eguchi, G. (1997) High-frequency generation of transgenic zebrafish which reliably express GFP in whole muscles or the whole body by using promoters of zebrafish origin. Dev. Biol. 192, 289–299.
17. Wixon, J. (2000) Featured organism: Danio rerio, the zebrafish. Yeast 17, 225–31.
18. Dodd, A., Curtis, P. M., Williams, L. C., and Love, D. R. (2000) Zebrafish: Bridging the gap between development and disease. Hum. Mol. Genet. 9, 2443–2449.
19. Haffter, P., Granato, M., Brand, M., Mullins, M. C., Hammerschmidt, M., Kane, D. A., Odenthal, J., van Eeden, F. J., Jiang, Y. P., Heisenberg, C. P., et al. (1996) The identification of genes with unique and essential functions in the development of the zebrafish, Danio rerio. Development 123, 1–36.
20. Mullins, M. C., Hammerschmidt, M., Haffter, P., and Nusslein-Volhard, C. (1994) Large-scale mutagenesis in the zebrafish: in search of genes controlling development in a vertebrate. Curr. Biol. 4, 189–202.
21. Amsterdam, A. and Hopkins, N. (2006) Mutagenesis strategies in zebrafish for identifying genes involved in development and disease. Trends Genet. 22, 473–478.
22. Peterson, R. T., Link, B. A., Dowling, J. E., and Schreiber, S. L. (2000) Small molecule developmental screens reveal the logic and timing of vertebrate development. Proc. Natl. Acad. Sci. USA 97,12965–12969.
23. Peterson, R. T., Shaw, S. Y., Peterson, T. A., Milan, D. J. Zhong, T. P., Schreiber, S. L., MacRae, C. A., and Fishman, M. C. (2004) Chemical suppression of a genetic mutation in a zebrafish model of aortic coarctation. Nat. Biotechnol. 22, 595–599.
24. Shafizadeh, E., Peterson, R. T., and Lin, S. (2004) Induction of reversible hemolytic anemia in living zebrafish using a novel small molecule. Comp. Biochem. Physiol. C Toxicol. Pharmacol. 138, 245–249.
25. Mathew, L. K., Andreasen, E. A., Sengupta, S., Peterson, R. T., and Tanguay, R. L. (nd) Chemical genetics to probe regenerative biology. J. Biol. Chem. 282, 35202–35210.
26. Nasevicius, A., and Ekker, S. C. (2001) The zebrafish as a novel system for functional genomics and therapeutic development applications. Curr. Opin. Mol. Ther. 3, 224–228.
27. Nasevicius, A., and Ekker, S. C. (2000) Effective targeted gene ‘knockdown’ in zebrafish. Nat. Genet. 26, 216–220.
28. Baumann, M., and Sander, K. (1984) Bipartite axiation follows incomplete epiboly in zebrafish embryos treated with chemical teratogens. J. Exp. Zool. 230, 363–376.
29. Carvan, M. J., 3rd, Loucks, E., Weber, D. N., and Williams, F. E. (2004) Ethanol effects on the developing zebrafish: Neurobehavior and skeletal morphogenesis. Neurotoxicol. Teratol. 26, 757–768.
30. Laale, H. W. (1977) The biology and use of zebrafish, Brachydanio rerio, in fisheries research. A literature review. J. Fish Biol. 10, 121–173.
31. Loucks, E. and Carvan, M. J., 3rd. (2004) Strain-dependent effects of developmental ethanol exposure in zebrafish. Neurotoxicol. Teratol. 26, 745–755.
32. Reimers, M. J., La Du, J. K., Periera, C. B., Giovanini, J., and Tanguay, R. L. (2006) Ethanol-dependent toxicity in zebrafish is partially attenuated by antioxidants. Neurotoxicol. Teratol. 28, 497–508.
33. Reimers, M. J., Hahn, M. E., and Tanguay, R. L. (2004) Two zebrafish alcohol dehydrogenases share common ancestry with mammalian class I, II, IV, and V alcohol dehydrogenase genes but have distinct functional characteristics. J. Biol. Chem. 279, 38303–38312.
34. Lassen, N., Estey, T., Tanguay, R. L., Pappa, A., Reimers, M. J., and Vasiliou, V. (2005) Molecular cloning, baculovirus expression, and tissue distribution of the zebrafish aldehyde dehydrogenase 2. Drug Metab. Dispos. 33, 649–656.
35. Jowett, T., and Lettice, L. (1994) Whole-mount in situ hybridizations on zebrafish embryos using a mixture of digoxigenin- and fluorescein-labelled probes. Trends Genet. 10, 73–74.
36. Linney, E., and Udvadia, A. J. (2004) Construction and detection of fluorescent, germline transgenic zebrafish. Methods Mol. Biol. 254, 271–288.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2008 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Tanguay, R.L., Reimers, M.J. (2008). Analysis of Ethanol Developmental Toxicity in Zebrafish. In: Nagy, L.E. (eds) Alcohol. Methods in Molecular Biology™, vol 447. Humana Press. https://doi.org/10.1007/978-1-59745-242-7_5
Download citation
DOI: https://doi.org/10.1007/978-1-59745-242-7_5
Publisher Name: Humana Press
Print ISBN: 978-1-58829-906-2
Online ISBN: 978-1-59745-242-7
eBook Packages: Springer Protocols