A standardized method for the three-way differential staining of plant chromosomes and the scoring of SCEs per cell cycle

We have made use of 2 alternative methodologies to obtain 3-way differential staining (TWD) in third-mitosis (M 3) chromosomes of Allium cepa, which involve different uptakes of bromodeoxyuridine (BrdU) through 3 cell divisions, in order to evaluate the sister-chromatid exchanges (SCE) frequency on...

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Bibliographic Details
Published inMutation Research/Environmental Mutagenesis and Related Subjects Vol. 216; no. 4; pp. 203 - 209
Main Authors Escalza, P., Piñero, J., Cortés, F.
Format Journal Article
LanguageEnglish
Published Elsevier B.V 1989
Subjects
Allium cepa
Plant chromosomes
SCE frequency
Three-way differential staining
Allium cepa
Plant chromosomes
Three-way differential staining
SCE frequency
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Summary:We have made use of 2 alternative methodologies to obtain 3-way differential staining (TWD) in third-mitosis (M 3) chromosomes of Allium cepa, which involve different uptakes of bromodeoxyuridine (BrdU) through 3 cell divisions, in order to evaluate the sister-chromatid exchanges (SCE) frequency on a per-cell-cycle basis. The main innovation introduced by us to the original protocols previously reported has been the use of 5-fluorodeoxyuridine (FdU) to inhibit endogenous thymidine (dT) synthesis. By using different [BrdU]: [dT] ratios in the presence of FdU the relative incorporation of BrdU into replicating DNA can be controlled. The use of 2 different approaches to obtain 3-way differentiation of sister chromatids allowed us to evaluate the role of BrdU in the induction of SCEs in our system. Both procedures rendered nearly 100% of M 3 chromosomes showing TWD. An additional advantage of our methodologies is their high degree of reproducibility.
ISSN:0165-1161
DOI:10.1016/0165-1161(89)90006-X