Skip to main content
SpringerLink
Log in

Zebrafish as a Model for the Study of Solid Malignancies

  • Protocol
  • First Online:
Zebrafish

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1451))

Abstract

Zebrafish cancer models have provided critical insight into understanding the link between aberrant developmental pathways and tumorigenesis. The unique strengths of zebrafish as compared to other vertebrate model systems include the combination of fecundity, readily available and efficient transgenesis techniques, transparency that facilitates in vivo cell lineage tracing, and amenability for high-throughput applications. In addition to early embryo readouts, zebrafish can develop tumors at ages ranging from 2 weeks old to adulthood. Tumorigenesis is driven by genetically introducing oncogenes using selected promoter/tissue-specific expression, with either mosaic expression or with the generation of a stable transgenic line. Here, we detail a research pipeline to facilitate the study of human oncogenes in zebrafish systems. The goals of this approach are to identify conserved developmental pathways that may be critical for tumor development and to create platforms for testing novel therapies.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 99.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 129.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 179.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Smolowitz R, Hanley J, Richmond H (2002) A three-year retrospective study of abdominal tumors in zebrafish maintained in an aquatic laboratory animal facility. Biol Bull 203(2):265–266

    Article  PubMed  Google Scholar 

  2. Kent ML, Spitsbergen JM, Matthews JM, Fournie JW, Murray KN, Westerfield M Diseases of zebrafish in research facilities. In ZIRC Health Services zebrafish disease manual 2012. http://zebrafish.org/health/diseaseManual.php

  3. Stanton MF (1965) Diethylnitrosamine-induced hepatic degeneration and neoplasia in the aquarium fish Brachydanio Rerio. J Natl Cancer Inst 34:117–130

    CAS  PubMed  Google Scholar 

  4. Pliss GB, Khudoley VV (1975) Tumor induction by carcinogenic agents in aquarium fish. J Natl Cancer Inst 55(1):129–136

    CAS  PubMed  Google Scholar 

  5. Feitsma H, Cuppen E (2008) Zebrafish as a cancer model. Mol Cancer Res 6(5):685–694

    Article  CAS  PubMed  Google Scholar 

  6. Berghmans S et al (2005) tp53 mutant zebrafish develop malignant peripheral nerve sheath tumors. Proc Natl Acad Sci U S A 102(2):407–412

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Langenau DM et al (2003) Myc-induced T cell leukemia in transgenic zebrafish. Science 299(5608):887–890

    Article  CAS  PubMed  Google Scholar 

  8. Langenau DM et al (2005) Cre/lox-regulated transgenic zebrafish model with conditional myc-induced T cell acute lymphoblastic leukemia. Proc Natl Acad Sci U S A 102(17):6068–6073

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Sabaawy HE et al (2006) TEL-AML1 transgenic zebrafish model of precursor B cell acute lymphoblastic leukemia. Proc Natl Acad Sci U S A 103(41):15166–15171

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Zhuravleva J et al (2008) MOZ/TIF2-induced acute myeloid leukaemia in transgenic fish. Br J Haematol 143(3):378–382

    Article  CAS  PubMed  Google Scholar 

  11. Yang HW et al (2004) Targeted expression of human MYCN selectively causes pancreatic neuroendocrine tumors in transgenic zebrafish. Cancer Res 64(20):7256–7262

    Article  CAS  PubMed  Google Scholar 

  12. Santoriello C et al (2010) Kita driven expression of oncogenic HRAS leads to early onset and highly penetrant melanoma in zebrafish. PLoS One 5(12):e15170

    Article  PubMed  PubMed Central  Google Scholar 

  13. Patton EE et al (2005) BRAF mutations are sufficient to promote nevi formation and cooperate with p53 in the genesis of melanoma. Curr Biol 15(3):249–254

    Article  CAS  PubMed  Google Scholar 

  14. Dovey M, White RM, Zon LI (2009) Oncogenic NRAS cooperates with p53 loss to generate melanoma in zebrafish. Zebrafish 6(4):397–404

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Langenau DM et al (2007) Effects of RAS on the genesis of embryonal rhabdomyosarcoma. Genes Dev 21(11):1382–1395

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Leacock SW et al (2012) A zebrafish transgenic model of Ewing’s sarcoma reveals conserved mediators of EWS-FLI1 tumorigenesis. Dis Model Mech 5(1):95–106

    Article  CAS  PubMed  Google Scholar 

  17. Gutierrez A et al (2011) Aberrant AKT activation drives well-differentiated liposarcoma. Proc Natl Acad Sci U S A 108(39):16386–16391

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Nguyen AT et al (2012) An inducible kras(V12) transgenic zebrafish model for liver tumorigenesis and chemical drug screening. Dis Model Mech 5(1):63–72

    Article  CAS  PubMed  Google Scholar 

  19. Neumann JC et al (2011) Mutation in the type IB bone morphogenetic protein receptor Alk6b impairs germ-cell differentiation and causes germ-cell tumors in zebrafish. Proc Natl Acad Sci U S A 108(32):13153–13158

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Howe K et al (2013) The zebrafish reference genome sequence and its relationship to the human genome. Nature 496(7446):498–503

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Hwang WY et al (2013) Efficient genome editing in zebrafish using a CRISPR-Cas system. Nat Biotechnol 31(3):227–229

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Hruscha A et al (2013) Efficient CRISPR/Cas9 genome editing with low off-target effects in zebrafish. Development 140(24):4982–4987

    Article  CAS  PubMed  Google Scholar 

  23. Suster ML et al (2009) Transgenesis in zebrafish with the tol2 transposon system. Methods Mol Biol 561:41–63

    Article  CAS  PubMed  Google Scholar 

  24. Kawakami K, Shima A, Kawakami N (2000) Identification of a functional transposase of the Tol2 element, an Ac-like element from the Japanese medaka fish, and its transposition in the zebrafish germ lineage. Proc Natl Acad Sci U S A 97(21):11403–11408

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Novakovic B (1994) U.S. childhood cancer survival, 1973–1987. Med Pediatr Oncol 23(6):480–486

    Article  CAS  PubMed  Google Scholar 

  26. Martin EA et al (2012) Tadalafil alleviates muscle ischemia in patients with Becker muscular dystrophy. Sci Transl Med 4(162):162

    Article  Google Scholar 

  27. Kawahara G et al (2011) Drug screening in a zebrafish model of Duchenne muscular dystrophy. Proc Natl Acad Sci U S A 108(13):5331–5336

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. North TE et al (2007) Prostaglandin E2 regulates vertebrate haematopoietic stem cell homeostasis. Nature 447(7147):1007–1011

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Amatruda JF et al (2002) Zebrafish as a cancer model system. Cancer Cell 1(3):229–231

    Article  CAS  PubMed  Google Scholar 

  30. Lawrence MS et al (2014) Discovery and saturation analysis of cancer genes across 21 tumour types. Nature 505(7484):495–501

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Kwan KM et al (2007) The Tol2kit: a multisite gateway-based construction kit for Tol2 transposon transgenesis constructs. Dev Dyn 236(11):3088–3099

    Article  CAS  PubMed  Google Scholar 

  32. Manoli M, Driever W (2012) Fluorescence-activated cell sorting (FACS) of fluorescently tagged cells from zebrafish larvae for RNA isolation. Cold Spring Harb Protoc 2012(8)

    Google Scholar 

  33. Moore JL et al (2002) Fixation and decalcification of adult zebrafish for histological, immunocytochemical, and genotypic analysis. Biotechniques 32(2):296–298

    CAS  PubMed  Google Scholar 

  34. Scheer N, Campos-Ortega JA (1999) Use of the Gal4-UAS technique for targeted gene expression in the zebrafish. Mech Dev 80(2):153–158

    Article  CAS  PubMed  Google Scholar 

  35. Asakawa K, Kawakami K (2008) Targeted gene expression by the Gal4-UAS system in zebrafish. Dev Growth Differ 50(6):391–399

    Article  CAS  PubMed  Google Scholar 

  36. Kawakami K et al (2010) zTrap: zebrafish gene trap and enhancer trap database. BMC Dev Biol 10:105

    Article  PubMed  PubMed Central  Google Scholar 

  37. Mosimann C et al (2011) Ubiquitous transgene expression and Cre-based recombination driven by the ubiquitin promoter in zebrafish. Development 138(1):169–177

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Burger A et al (2014) A zebrafish model of chordoma initiated by notochord-driven expression of HRASV12. Dis Model Mech 7(7):907–913

    Article  PubMed  Google Scholar 

  39. Amsterdam A et al (2004) Many ribosomal protein genes are cancer genes in zebrafish. PLoS Biol 2(5):E139

    Article  PubMed  PubMed Central  Google Scholar 

  40. Jungke P, Hans S, Brand M (2013) The zebrafish CreZoo: an easy-to-handle database for novel CreER(T2)-driver lines. Zebrafish 10(3):259–263

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

GCK is supported by a Cancer Prevention and Research Institute of Texas postdoctoral fellowship through the UTSW Cancer Intervention and Prevention Discoveries training program, a QuadW-American Association for Cancer Research Fellowship for Clinical/Translational Sarcoma Research, and a Young Investigator Grant from Alex’s Lemonade Stand Foundation. Supported by grants R01CA135731 from the NIH and RP120685 from the Cancer Prevention and Research Institute of Texas (to JFA). JFA is supported by the Nearburg Family Professorship in Pediatric Oncology Research.

Author information

Authors and Affiliations

  1. Department of Pediatrics, UT Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX, 75390, USA

    Genevieve C. Kendall & James F. Amatruda

  2. Department of Molecular Biology, UT Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX, 75390, USA

    Genevieve C. Kendall & James F. Amatruda

  3. Department of Internal Medicine, UT Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX, 75390, USA

    James F. Amatruda

Authors
  1. Genevieve C. Kendall
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. James F. Amatruda
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to James F. Amatruda .

Editor information

Editors and Affiliations

  1. Division of Molecular and Developmental Biology, National Institute of Genetics, Mishima, Japan

    Koichi Kawakami

  2. Institute of Genetics and Molecular Medicine, MRC Human Genetics Unit & The University of Edinburgh Cancer Research UK Centre, Edinburgh, United Kingdom

    E. Elizabeth Patton

  3. Champalimaud Centre for the Unknown, Champalimaud Neuroscience Programme Champalimaud Centre for the Unknown, Lisbon, Portugal

    Michael Orger

Rights and permissions

Reprints and permissions

Copyright information

© 2016 Springer Science+Business Media New York

About this protocol

Cite this protocol

Kendall, G.C., Amatruda, J.F. (2016). Zebrafish as a Model for the Study of Solid Malignancies. In: Kawakami, K., Patton, E., Orger, M. (eds) Zebrafish. Methods in Molecular Biology, vol 1451. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3771-4_9

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-3771-4_9

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3769-1

  • Online ISBN: 978-1-4939-3771-4

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation