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A method for the analysis of sister diromatid exchanges (SCEs) occurring in thrce subsequent cell divisions in uivo, was developed in murine bone marrow cells. The method was based on the three-way differential staining of sister chromatids. Mice were injected twice ¡.p. with BrdU adsorbed to activated charcoal; the fint dose at O h (fint cell cycle) was varied between 0.1 and 0.25 mg of BrdU/g of body weight and the second dose at 8 h (second and third cell cycles), was either 1.0, 1.33 or 1.5 mg of BrdU/g of body weight. Animals were injected with colchicine (¡.p.) two houn before sacrificing and chromosome prepations were obtained by the usual method. After sister chromatid differentiation by the method of fluomscence plus Giemsa the cells were scored for well and poor differential staining and in some cases for SCE frequency in three subsequent cell divisions. The results indicate that a higher percentage of well-differentiated three way stained cells were obtained with the combination of BrdU doses of 0.2 and 1.5 mg/g of body weight. The very low basal frequency of SCE observed in the first cell cycle (0.31-0.46) probably increases the sensitivity to detected the mutagenic effect by SCE method. This method can be used for both the analysis of in uiuo penistence of lesions involved in SCE in,duction as well as the detection of sinergism between the mutagen action and the BrdU incorporatioa to DNA.

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Universidad Autónoma del Estado de México
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Versión 3.0 | 2017
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