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Approximation of baseline and BrdU-induced SCE frequencies

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Abstract

In the present paper we have developed a new rationale and an experimental schedule to approximate the frequency of SCEs which occur independently of BrdU incorporation, namely, the baseline frequency of SCEs. The method used includes the analysis of SCE yields in second and third division chromosomes after BrdU-substitution for 1, 2, and/or 3 successive replication rounds in the presence of this thymidine analogue, leading to a set of ten different experimental results. As a result of formulating various mathematical equations and applying them to the data, an accurate estimation of the frequency of baseline (BrdU-independent) and BrdU-induced SCEs, can be made, thus avoiding the difficulties inherent in the current extrapolation methods. The conclusions are that 1) SCEs seem to be formed after DNA synthesis (by exchanging post-replicative DNA portions), but, obviously, very near to the replication fork and 2) that under our experimental conditions about 0.065 SCEs per picogram of DNA per cell cycle occur as a consequence of chromosome replication, this frequency being increased by BrdU-substitution. The methodology seems to be reliable enough to be used in other species and systems in order to compare baseline SCE frequencies.

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Abbreviations

SCEs:

sister-chromatid exchanges

BrdU(BrdUrd):

5-bromodeoxyuridine

dTh(dThd):

thymidine

3H-dTh(3H-dThd):

tritiated thymidine

FdU(FdUrd):

5-fluorodeoxyuridine

Urd:

uridine

FPG:

fluorescent plus Giemsa

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Authors and Affiliations

  1. Instituto de Biología Celular (CSIC) Velázquez, 144, Madrid-6, Spain

    C. Gutiérrez

  2. Facultad de Ciencias, Universidad de Córdoba, Córdoba, Spain

    A. Calvo

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  1. C. Gutiérrez
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  2. A. Calvo
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Gutiérrez, C., Calvo, A. Approximation of baseline and BrdU-induced SCE frequencies. Chromosoma 83, 685–695 (1981). https://doi.org/10.1007/BF00328527

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