For decades, spontaneous and induced mutant animals have yielded valuable insights into the roles of specific genes in development and disease. Recently, transgenic and knockout mice, together with large-scale mutagenesis screens of model organisms such as the fruitfly, Drosophila, and the nematode, C. elegans, have contributed detailed insights into the roles of specific genes in development. These intensive studies reveal a remarkable degree of conservation of genetic strategies for control of cell differentiation, cell proliferation, and cell death, as well as pattern formation during development throughout all animals. Advantages of zebrafish for study of fundamental vertebrate genetic principles and molecular developmental genetics include small adult size, relative hardiness in small aquaria, high fecundity, external fertilization and development with accessibility of all developmental stages, highly transparent eggshells, rapid embryonic development, short generation times, ease of embryonal cloning to allow gynogenesis or androgenesis, and applicability for large-scale, rapid mutant screens. Intense worldwide research in zebrafish molecular development, genomics, and mutagenesis during the past decade has complemented the previous strong database on biology, physiology, and development of zebrafish creating a powerful system in which to answer fundamental questions in biology and medicine. The investigators will develop lines of zebrafish highly sensitive to specific histological types of neoplasia occurring in specific tissues. To develop these lines, the investigators will evaluate carcinogen-induced neoplasia in lines of zebrafish with inactivating mutations in orthologs of human tumor suppressor genes. The investigators will screen offspring of fish mutagenized with N-ethyl-N-nitrosourea (ENU), using temperature mediated heteroduplex analysis (TMHA) and the mismatch cleavage enzyme CELI to detect point mutations in zebrafish orthologs of the human tumor suppressor genes P53, RB1, WT1, PTEN, or NF1. The investigators will treat zebrafish embryos or fry of these mutant lines by immersion exposure to N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) or 7,12-dimethylbenz[a]anthracene (DMBA), then will evaluate neoplasm incidences six and 12 months later. These fish will provide a rapid and relatively inexpensive whole-animal bioassay system to detect carcinogenic risks to man from new materials as well as from environmental contaminants. These lines of zebrafish will provide an efficient and inexpensive system for development of new treatment and prevention strategies for cancer, and will help identify new genes controlling growth and differentiation.

Agency
National Institute of Health (NIH)
Institute
National Institute of Environmental Health Sciences (NIEHS)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21ES013124-02
Application #
6931960
Study Section
Special Emphasis Panel (ZRR1-CM-9 (02))
Program Officer
Heindel, Jerrold
Project Start
2004-08-01
Project End
2008-04-30
Budget Start
2005-05-01
Budget End
2008-04-30
Support Year
2
Fiscal Year
2005
Total Cost
$141,500
Indirect Cost
Name
Oregon State University
Department
Public Health & Prev Medicine
Type
Schools of Earth Sciences/Natur
DUNS #
053599908
City
Corvallis
State
OR
Country
United States
Zip Code
97339
Cooper, T K; Spitsbergen, J M (2016) Valvular and Mural Endocardiosis in Aging Zebrafish (Danio rerio). Vet Pathol 53:504-9
Cooper, T K; Murray, K N; Spagnoli, S et al. (2015) Primary intestinal and vertebral chordomas in laboratory zebrafish (Danio rerio). Vet Pathol 52:388-92
Spitsbergen, Jan M; Buhler, Donald R; Peterson, Tracy S (2012) Neoplasia and neoplasm-associated lesions in laboratory colonies of zebrafish emphasizing key influences of diet and aquaculture system design. ILAR J 53:114-25
Balla, Keir M; Lugo-Villarino, Geanncarlo; Spitsbergen, Jan M et al. (2010) Eosinophils in the zebrafish: prospective isolation, characterization, and eosinophilia induction by helminth determinants. Blood 116:3944-54
Zhan, Huiqing; Spitsbergen, Jan M; Qing, Wei et al. (2010) Transgenic expression of walleye dermal sarcoma virus rv-cyclin gene in zebrafish and its suppressive effect on liver tumor development after carcinogen treatment. Mar Biotechnol (NY) 12:640-9
Ju, Bensheng; Spitsbergen, Jan; Eden, Christopher J et al. (2009) Co-activation of hedgehog and AKT pathways promote tumorigenesis in zebrafish. Mol Cancer 8:40
Spitsbergen, Jan M; Blazer, Vicki S; Bowser, Paul R et al. (2009) Finfish and aquatic invertebrate pathology resources for now and the future. Comp Biochem Physiol C Toxicol Pharmacol 149:249-57