Indian Journal of Science and Technology

Open Access Open Access  Restricted Access Subscription Access

The Usefulness of Two-colour Fluorescence in situ Hybridization Technique Using Chromosomal Subset at Metaphase and Interphase Stage in Radiation Biodosimetry


Affiliations
1 Department of Radiobiology, Institute for Environmental Sciences, Hachazawa 2-121,Takahoko, Rokkasho, Kamikita, Aomori 039-3213, Japan
2 Department of Cancer Cytogenetics, Research Institute for Radiation biology and Medicine, Hiroshima University, Kasumi-1-2-3, Minami-ku Hiroshima, Japan
3 Atomic Bomb Survivors Relief Foundation, Kurakake, Asakita-ku, Hiroshima 739-1743, India
 

Peripheral bloods from healthy donors were irradiated by 60Co γ-rays and californium (252Cf) -neutron for 0.5 to 4 Gy and the frequencies of the translocations were detected by two colour fluorescence in situ hybridization (FISH). Chromosomes 1, 2 and 3 and chromosomes 6, 7 and 9 were labeled by rodamine-dUTP and fluorescein-dUTP as viewing red or green signal on metaphase chromosomes, respectively. In separate observation, translocation was detected on the interphase cells by the two color FISH method using whole chromosome painting probes of chromosomes 4 and 9. Detected translocations on the metaphase and interphase cells showed clear dose response relationships. Translocations induced by 60Co γ -rays, 252Cf neutrons, and those in atomic bomb survivors were able to detect easily by both interphase FISH and metaphase FISH using two colour of chromosome subsets. These results suggest that scoring of translocations by two colour FISH of chromosome subsets can serve as more sensitive and applicable biological dosimeter for short or long-term radiation effects.

Keywords

Interphase FISH, Biodosimetry, Atomic Bomb, Neutron, γ Rays
User

  • Awa AA, Sofuni T, Honda T, Itoh T, Neriishi S and Ohtake M (1978) Relationship between the radiation dose and chromosome aberrations in atomic bomb survivors of Hiroshima and Nagasaki. J. Radiat. Res.119, 126-140.

  • Buckton KE (1983) Chromosome aberrations in patients treated with X-irradiation for ankylosing spondylitis. In: Radiation-induced Chromosome Damage in Man. (Eds.) Ishihara T & Sasaki MS, Alan R. Liss, New York, pp:491-511.

  • Cremer T and Cremer C (2001) Chromosome territories, nuclear architecture and gene regulation in mammalian cells. Nat. Rev. Genet. 2, 292-301.

  • Durante M, Gerge K, and Yang TC (1996) Biological dosimetry by interphase chromosome painting. Radiat. Res. 145, 53-60.

  • Durante M., Gerge K. and Yang TC (1997) Biodosimetry of ionizing radiation by selective painting of prematurely condensed chromosomes in human lymphocytes. Radiat. Res. 148, S45-S50.

  • Eils R, Bertin E, Saracoglu K, Rinke B, Schroeck E, Parazza F, Unch Y, Robert-Nicoud M, Stelzer EHK, Chassery JM, Cremer T and Cremer C (1995) Application of confocal laser microscopy and threedimensional Voronoi of interphase chromosomes. J. of Microscopy. 177, 150-161.

  • Figgitt M and Savage JR (1999) Interphase chromosome domain re-organization following irradiation. Int. J. Radiat. Biol. 75, 811-817.

  • Hoshi M, Matsuura M., Hayakawa N and Ito C (1993) A dose estimation system (ABS93D) for the atomic survivors in Hiroshima and its application. J. Radiat. Res. 34(4), 375, 1193-1200.

  • Hoshi M, Takeoka S, Tsujimura T, Kuroda T, Kawami M and Sawada S (1988) Dosimetric evaluation of 252Cf beam for use in radiobiology studies at Hiroshima University. Phys. Med. Biol. 33, 473-480.

  • IAEA (2001) Cytogenetic Analysis for Radiation Dose Assessment, A manual, Technical reports series no. 405, International Atomic Energy Agency, Vienna, pp:66-67.

  • Kamada N (1999) Biological dosimetry of atomic bomb survivors exposed within 500 meters form the hypocenter and health consequences. J. Radiat. Res. 40, Suppl., 155-164.

  • Kamada N and Tanaka K (1983) Cytogenetic studies of hematological disorders in atomic bomb survivors. In: Radiation-induced Chromosome Damage in Man. (Eds) Ishihara T and Sasaki MS, Alan R. Liss, New York, pp:455-474.

  • Kamada N, Tanaka K, Asou H, Toge T, Kuramoto A, Hoshi M, Matsuura M, Hayakawa N and Ito C (1994) Synthetic medical studies on atomic bomb survivors exposed in short distance. XXI Dose estimation based on ABS93D by means of chromosome aberration rate. Hiroshima Med. Acc. 47(3) 430-432. (in Japanese)

  • Kanda R, Hayata I and Lloyd DC (1999) Easy biodosoimetry for high –dose radiation exposures using drug-induced, prematurely condensed chromosomes. Int. J. Radiat. Biol. 75, 441-446.

  • Leatherbarrow EL, Harper JV, Cucinotta FA and O’Neill P (2006) Induction and quantification of γ- H2AX foci following low and high LET-irradiation. Int. J. Radiat. Biol. 82, 111-118.

  • Liu H, Tanaka K and Kamada N (2008) Differential expression of PML in 60Coγ-ray andγ-IFN-induced apoptosis in B-lymphocytes. Indian J. Sci. Technol. 1(3), 1-13. http://www.indjst.org

  • Löbrich M, Reif N, Kühne M, Hechman, flecknenstein J, Rübe C and Uder M (2005) In vivo formation of and repair of DNA double-strand breaks after computed tomography examinations. Proc. Natl. Acad. Sci., USA 102, 8984-8989.

  • Lucas JN and Cervantes E (2002) Significant large scale chromosome territory movement occurs as a result of mitosis, but not during interphase. Int. J. Radiat. Biol. 78(6), 449-455.

  • Lucus JN, Awa A, Straume T, Poggensee M, Kodama Y, Nakano M, Ohtaki K, Weier H-U, Pinkel WD, Gray J and Littlefield G (1992) Rapid translocation frequency analysis in humans decades after exposure to ionizing radiation. Int. J. Radiat. Biol. 62, 53-63.

  • Lucus JN, Poggensee M and Straume T (1993) Translocations between two specific human chromosomes detected by three-color‘chromosome painting’ Cytogenet. Cell Genet. 62, 11-12.

  • M’kacher R Violet D, Aubert b, Girinsky T, Dossou J, Béron-Gaillard N, Carde P and Parmentier C (2003) Premature chromosome condensation associated with fluorescence in situ hybridization detects cytogenetic abnormalities after a CT scan: evaluation of the low-dose effects. Radiat. Prot. Dosimetry 103, 35-40.

  • Manders EMM, Visser AE, Koppen A, de Leeuw WC, van Liere R, Brakenhoff GI and van Driel R (2003) Four-dimentional imaging of chromation dynamics during the assembly of the interphase nucleus. Chromosome Res. 11, 537-547.

  • Manuelidis L (1985) Individual interphase chromosome domains reveled by in situ hybridization. Hum. Genet. 71, 288-293.

  • Morton (1991) Parameters of the human genome. Proc. Natl. Acad. Sci. USA. 88, 7474-7476.

  • Rinke B, Bischoff A, Meffert MC, Schrschmidt R, Hausman M, Stelzer EHK, Cremer T and Cremer C (1995) Volume ratios of painted chromosome territories 5, 7, and X in female human cell nuclei studied with confocal laser microscopy and the Cavalieri estimation. Bioimaging 3, 1-11.

  • Rupa DS, Hasegawa L and Eastmond DA (1995) Detection of chromosomal breakage in the 1cen-1q12 region on interphase human lymphocytes using multicolour fluorescence in situ hybridization with tandem DNA probes. Cancer Res. 55, 640-645.

  • Schardin M, Cremer T, Hager HD and Lang M (1985) Specific staining of human chromosomes in Chinese hamster × man hybrid cell lines demonstrate interphase chromosome territories. Hum. Genet.71, 281-287.

  • Tanabe H, Müller S, Nevesser M, Hase JH, Calcagno E, Cremer M, Solovei I, Cremer C and Cremer T (2002) Evolutionary conservation of chromosome territory arrangements in cell nuclei from higher primates. Proc. Natl. Acad. Sci. USA. 99, 4424-4429.

  • Tanaka K, Gajendran N and Kamada N (2009) Relative biological effectiveness (RBE) and dose rate dependent ratio of translocation to dicentric chromosome yield in 252Cf neutrons. Indian J. Sci. Technol. 2(1), 1-11. http://www.indjst.org.

  • Tanaka K, Kamada N, Kuramoto A and Ohkita T (1986) Chromosome aberrations in bone marrow in exposed atomic bomb survivors exposed at nearly hypocenter. Hiroshima Med. Acc. 39(2), 244-248. (in Japanese)

  • Tanaka K, Kamada N, Ohkita T and Kuramoto A (1983) Nonrandom distribution of chromosome breaks in lymphocytes of atomic bomb survivors. J. Radiat. Res.24 (4) 291-304.

  • Tanaka K, Popp S, Fisher C, Van Kaik G, Kamada N, Cremer T and Cremer C (1996) Chromosome aberration analysis in atomic bomb survivors and Thorotrast patients using two-and three-colour chromosome painting of chromosomal subsets. Int. J. Radiat. Biol. 70(1), 95-108.

  • Tucker JD, Morgan WF, Awa AA, Bauchinger M, Blakey D, Conforth MN, Littlefield LG, Natarajan AT and Shasserre C (1995) A proposed system for scoring structural aberrations detected by chromosome painting. Cytogenet. Cell Genet. 68, 211-222.

  • Vázquex-Gundín F, Rivero TM, Gosálvez J and Fernández LJ (2002) Radiation-induced DNA breaks in different human satellite DNA sequence areas, analyzed by DNA breakage detection-fluorescence in situ hybridization. Radiat. Res. 157, 711-720.


Abstract Views: 478

PDF Views: 84




  • The Usefulness of Two-colour Fluorescence in situ Hybridization Technique Using Chromosomal Subset at Metaphase and Interphase Stage in Radiation Biodosimetry

Abstract Views: 478  |  PDF Views: 84

Authors

Kimio Tanaka
Department of Radiobiology, Institute for Environmental Sciences, Hachazawa 2-121,Takahoko, Rokkasho, Kamikita, Aomori 039-3213, Japan
Mansyur Arif
Department of Cancer Cytogenetics, Research Institute for Radiation biology and Medicine, Hiroshima University, Kasumi-1-2-3, Minami-ku Hiroshima, Japan
Nanao Kamada
Atomic Bomb Survivors Relief Foundation, Kurakake, Asakita-ku, Hiroshima 739-1743, India

Abstract


Peripheral bloods from healthy donors were irradiated by 60Co γ-rays and californium (252Cf) -neutron for 0.5 to 4 Gy and the frequencies of the translocations were detected by two colour fluorescence in situ hybridization (FISH). Chromosomes 1, 2 and 3 and chromosomes 6, 7 and 9 were labeled by rodamine-dUTP and fluorescein-dUTP as viewing red or green signal on metaphase chromosomes, respectively. In separate observation, translocation was detected on the interphase cells by the two color FISH method using whole chromosome painting probes of chromosomes 4 and 9. Detected translocations on the metaphase and interphase cells showed clear dose response relationships. Translocations induced by 60Co γ -rays, 252Cf neutrons, and those in atomic bomb survivors were able to detect easily by both interphase FISH and metaphase FISH using two colour of chromosome subsets. These results suggest that scoring of translocations by two colour FISH of chromosome subsets can serve as more sensitive and applicable biological dosimeter for short or long-term radiation effects.

Keywords


Interphase FISH, Biodosimetry, Atomic Bomb, Neutron, γ Rays

References





DOI: https://doi.org/10.17485/ijst%2F2009%2Fv2i4%2F29422