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Single Cell Gel Electrophoretic Analysis of 2-Aminoanthracene Exposed F-344 Rats
American Journal of Biochemistry and Molecular Biology (2011)
  • Worlanyo E. Gato, Georgia Southern University
  • Jay C. Means, Southern Illinois University Carbondale
Abstract
To assess DNA damage from exposure to 2-Aminoanthracene (2AA), Single Cell Gel Electrophoresis (SGCE) was employed to examine single strand breaks in blood of Fisher-344 rats. Although subcellular injury due to 2-AA exposure has been noted in the past, there is yet to be a direct demonstration of genetic alterations due to 2-AA intoxication. In the current study, alkaline comet assay was used to evaluate the type of DNA damage that is on-going in the blood samples of F-344 animals. The animals were fed control (0 mg kg-1), LD (50 mg kg-1), MD (75 mg kg-1) and HD (100 mg kg-1) 2-AA diet for two and four weeks. At the end of each exposure period (14 or 28 days), rats were euthanized with CO2 and blood was collected by cardiac puncture. Twenty micro liters of whole blood samples were added to 1 mL of Hanks Balanced Salt Solution (HBSS without Ca2+ and Mg2+) and subsequently snapped frozen in liquid nitrogen. Following comet assay analysis, single strand break were assessed via tail moment, tail length, tail intensity and head intensity scoring of comets along with cell viability. There seems to be an apparent dose response. That is 50, 75 and 100 mg kg-1 diet 2-AA rats when compared with the control demonstrated significant (p<0.05) damage as measured by tail length and tail moment values. Head and tail intensity values for intoxicated animals were also significant at 2 and 4 weeks relative to the controls, although there were no dramatic shifts between exposure time.
Keywords
  • 2-Aminoanthracene (2-AA),
  • Comet assay,
  • Single strand break,
  • DNA damage
Publication Date
2011
DOI
10.3923/ajbmb.2011.275.283
Citation Information
Worlanyo E. Gato and Jay C. Means. "Single Cell Gel Electrophoretic Analysis of 2-Aminoanthracene Exposed F-344 Rats" American Journal of Biochemistry and Molecular Biology Vol. 1 Iss. 3 (2011) p. 275 - 283 ISSN: 2150-4210
Available at: http://works.bepress.com/worlanyo_gato/5/