Ret, Abl1 (cAbl) and Trp53 Gene Fragmentations in Comet-FISH Assay Act as In Vivo Biomarkers of Radiation Exposure in C57BL/6 and CBA/J Mice

• Published in: Radiation research 2006-05, Vol.165 (5), p.553-561
• Main Authors: Amendola, Roberto, Basso, Emiliano, Pacifici, Pier Giorgio, Piras, Elena, Giovanetti, Anna, Volpato, Claudia, Romeo, Giovanni
• Format: Article
• Language: English
• Subjects: Adaptor Proteins, Signal Transducing - genetics; Animals; Apoptosis; Biomarkers - analysis; Cells, Cultured; Comet Assay - methods; Comets; Cytoskeletal Proteins; DNA - genetics; DNA - radiation effects; DNA Damage; DNA Fragmentation - radiation effects; DNA probes; Environmental Exposure - analysis; In Situ Hybridization, Fluorescence - methods; Ionizing radiation; Irradiation; Leukocytes, Mononuclear - metabolism; Leukocytes, Mononuclear - radiation effects; Mice; Mice, Inbred C57BL; Mice, Inbred CBA; Proto-Oncogene Proteins c-ret - genetics; Radiation damage; Radiation dosage; Radiation genetics; Radiation Monitoring - methods; Tumor Suppressor Protein p53 - genetics
• ISSN: 0033-7587; 1938-5404
• DOI: 10.1667/RR3544.1

Amendola, R., Basso, E., Pacifici, P. G., Piras, E., Giovanetti, A., Volpato, C. and Romeo, G. Ret, Abl1 (cAbl) and Trp53 Gene Fragmentations in Comet-FISH Assay Act as In Vivo Biomarkers of Radiation Exposure in C57BL/6 and CBA/J Mice. Radiat. Res. 165, 553–561 (2006). The International Commission on Radiation Protection (ICRP) has lowered the dose limits for workers and for the general public exposed to ionizing radiation. Consequently, a reliable dosimetric method for monitoring possible radiation-induced damage is of great importance in radioprotection. The counting of dicentric chromosomal aberrations and of micronuclei in peripheral blood lymphocytes is unreliable when it is applied to in vivo biopsies and for low-dose exposures. Single-cell gel electrophoresis (SCGE or comet assay), although sensitive and rapid, shows high variability when applied in vivo, probably due to prompt repair of the DNA breaks and confounding environmental factors. In this paper, we describe specific in situ hybridization of Ret, Abl1 (cAbl), and Trp53 gene fragmentations on SCGE slides (comet-FISH assay) in peripheral blood cells from C57BL/6 and CBA/J mice as an indicator of radiation-induced DNA damage. The results obtained from four mice for each experimental point (0, 1, 2 and 4 Gy of X rays) discriminated in a statistically significant way the effects of all doses when fragmentations were analyzed for the Ret, Ab1 and Trp53 genes. SCGE alone, when applied to the same specimens, produced no significant results because of interindividual and experimental variability.