Zebrafish as a "biosensor"? Effects of ionizing radiation and amifostine on embryonic viability and development.

The zebrafish (Danio rerio) has emerged as a popular vertebrate model system for cancer and treatment-related research. Benefits include ease of care, rapid development, optical clarity of embryos, which allows visualization of major organ systems, and opportunities for genetic manipulation. However, specific parameters of radiation sensitivity have not been systematically documented. We investigated the effects of radiation and a radiomodifier on zebrafish viability and embryonic development. Embryos were exposed to gamma-radiation (5, 10, or 20 Gy) at sequential times postfertilization and serially assessed for viability and morphologic abnormalities. As expected, lethality and morphologic perturbations were more pronounced earlier in embryogenesis and with higher radiation doses and were partially reversed by amifostine. The effects of radiation and concurrent treatment with amifostine on the developmental organization of the eye and brain were striking. Radiation resulted in hypocellularity and disorganization of the cellular layers of the retina, effects partially reversed by amifostine, as well as lens opacification. Radiation strikingly reduced the volume of brain, but the volume loss was substantially blocked by amifostine. Increased terminal deoxyribonucleotidyl transferase-mediated dUTP nick end labeling signal was noted in both the irradiated eye and brain, but reduced by amifostine. Finally, irradiating embryos resulted in caspase activation detectable in 96-well microplates, which was proportional to the number of embryos and radiation dose; the degree of activation was markedly reduced by amifostine. These results together suggest the power and versatility of the zebrafish in assessing the effects of radiation and radiomodifiers on organ and tissue development.

[1]  Randall W King,et al.  Small molecules that delay S phase suppress a zebrafish bmyb mutant , 2005, Nature chemical biology.

[2]  M. McAleer,et al.  Coordinate control of cell cycle regulatory genes in zebrafish development tested by cyclin D1 knockdown with morpholino phosphorodiamidates and hydroxyprolyl-phosphono peptide nucleic acids , 2005, Nucleic acids research.

[3]  W. Dynan,et al.  DNA damage response and Ku80 function in the vertebrate embryo , 2005, Nucleic acids research.

[4]  B. Movsas,et al.  Amifostine in chemoradiation therapy for non-small cell lung cancer: review of experience and design of a phase II trial assessing subcutaneous and intravenous bolus administration. , 2005, Seminars in oncology.

[5]  N. Andratschke,et al.  Prevention of radiation-induced central nervous system toxicity: a role for amifostine? , 2004, Anticancer research.

[6]  S. Ekker,et al.  Zebrafish as a genomics research model. , 2004, Current pharmaceutical biotechnology.

[7]  Adam P Dicker,et al.  Novel use of zebrafish as a vertebrate model to screen radiation protectors and sensitizers. , 2004, International journal of radiation oncology, biology, physics.

[8]  Tae-hwan Kim,et al.  Dose-incidence relationships on the prenatal effects of gamma-radiation in mice. , 2002, Journal of veterinary science.

[9]  L. S. Ross,et al.  Apoptosis in the developing zebrafish embryo. , 2001, Developmental biology.

[10]  E. Ségal-Bendirdjian,et al.  Staurosporine induces apoptosis through both caspase-dependent and caspase-independent mechanisms , 2001, Oncogene.

[11]  R. DiPaola,et al.  Overview of phase I trials of multitargeted antifolate (MTA, LY231514). , 1999 .

[12]  R. Siden,et al.  Amifostine for protection from antineoplastic drug toxicity. , 1997, American journal of health-system pharmacy : AJHP : official journal of the American Society of Health-System Pharmacists.

[13]  R. Capizzi.  Amifostine: the preclinical basis for broad-spectrum selective cytoprotection of normal tissues from cytotoxic therapies. , 1996, Seminars in oncology.

[14]  J. Crawford.  Update: vinorelbine (navelbine) in non-small cell lung cancer. , 1996, Seminars in oncology.

[15]  M. Inouye,et al.  Effects of different doses of gamma-radiation on the developing brain of mice. , 1995, Environmental medicine : annual report of the Research Institute of Environmental Medicine, Nagoya University.

[16]  C. Kimmel,et al.  Stages of embryonic development of the zebrafish , 1995, Developmental dynamics : an official publication of the American Association of Anatomists.

[17]  K. Kohn,et al.  Induction of a common pathway of apoptosis by staurosporine. , 1994, Experimental cell research.

[18]  H. Lassmann,et al.  Detection of DNA fragmentation in apoptosis: application of in situ nick translation to cell culture systems and tissue sections. , 1993, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[19]  S. Ben‐Sasson,et al.  Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation , 1992, The Journal of cell biology.

[20]  G. Steel,et al.  Oxygen-dependent protection of radiation lung damage in mice by WR 2721. , 1984, International journal of radiation biology and related studies in physics, chemistry, and medicine.

[21]  C. D'amato,et al.  Effects of radiation on development, especially of the nervous system. , 1980, The American journal of forensic medicine and pathology.

[22]  E. W. Hancock,et al.  RADIATION-INDUCED HEART DISEASE. A STUDY OF TWENTY-FIVE PATIENTS. , 1967 .

[23]  K G Johnson,et al.  In utero exposure to the Hiroshima atomic bomb. An evaluation of head size and mental retardation: twenty years later. , 1967, Pediatrics.

[24]  S. Kimura,et al.  Atomic-bomb radiation cataract. Case report with histopathologic study. , 1951, American journal of ophthalmology.

[25]  J Yuan,et al.  Caspases: an ancient cellular sword of Damocles , 2004, Cell Death and Differentiation.

[26]  C. Grau,et al.  Chemical radioprotection: a critical review of amifostine as a cytoprotector in radiotherapy. , 2003, Seminars in radiation oncology.

[27]  Michael Brand,et al.  Keeping and raising zebrafish , 2002 .

[28]  Ralf Dahm,et al.  Zebrafish: A Practical Approach. Edited by C. NÜSSLEIN-VOLHARD and R. DAHM. Oxford University Press. 2002. 322 pages. ISBN 0 19 963808 X. Price £40.00 (paperback). ISBN 0 19 963809 8. Price £80.00 (hardback). , 2003 .

[29]  R. DiPaola,et al.  Neurologic protection by amifostine. , 1999, Seminars in oncology.

[30]  M. Westerfield.  The zebrafish book : a guide for the laboratory use of zebrafish (Danio rerio) , 1995 .

[31]  S. Roberts,et al.  Radiation-induced ocular injury in the dog: a histological study. , 1990, International journal of radiation oncology, biology, physics.

[32]  F. Giblin,et al.  Effect of radioprotective agents on X-ray cataracts. , 1989, Lens and eye toxicity research.

[33]  G. Streisinger,et al.  INDUCTION OF MUTATIONS BY γ-RAYS IN PREGONIAL GERM CELLS OF ZEBRAFISH EMBRYOS , 1983 .

[34]  R. Stroobandt,et al.  Radiation-induced heart disease. , 1975, Acta cardiologica.

[35]  L. F. Fajardo,et al.  Radiation-induced heart disease. Clinical and experimental aspects. , 1971, Radiologic clinics of North America.