Analysis of chromosome replication by a BrdU antibody technique

Chromosome replication was studied without synchronization in human lymphocyte and amniotic cell cultures visualizing very short 5-bromodeoxyuridine (BrdU) pulses by an immunologic technique (BAT). The findings agree in general with those facts known from earlier BrdU staining techniques. The very h...

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Bibliographic Details
Published in:Chromosoma 1989-11, Vol.98 (5), p.335-341
Main Authors: VOGEL, W, AUTENRIETH, M, MEHNERT, K
Format: Article
Language:English
Subjects:
5-bromodeoxyuridine
amnion
Azure Stains
Biological and medical sciences
Bromouracil
Cells, Cultured
Chromosome Banding
Chromosomes - physiology
Chromosomes - ultrastructure
Cytogenetics
DNA - biosynthesis
DNA Replication
Fundamental and applied biological sciences. Psychology
Genetics of eukaryotes. Biological and molecular evolution
Human
Humans
Interphase
Karyotyping
lymphocytes
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Summary:Chromosome replication was studied without synchronization in human lymphocyte and amniotic cell cultures visualizing very short 5-bromodeoxyuridine (BrdU) pulses by an immunologic technique (BAT). The findings agree in general with those facts known from earlier BrdU staining techniques. The very high sensitivity of BAT was shown to allow the detection of replication in a band where 1 in 200 nucleotides is replaced by BrdU. The main observations are: though the replication patterns after BAT appear strange the bands correspond to those described by the Paris Conference (1971). At the beginning of the S-phase a stepwise onset of replication in only a subset of R-bands is confirmed. There is a considerable difference in the sensitivity between early and late S (SE and SL) for the detection of BrdU pulses. This difference probably reflects a different spatial arrangement of chromatin in R-bands as compared with G-bands below the level of cytogenetic analysis. The use of short pulses did not reveal any additional subdivision of SE or SL. The correspondence between chromosomal bands and replicon clusters is discussed briefly with respect to the different time they need for replication.
ISSN:0009-5915
1432-0886
DOI:10.1007/bf00292386