PMID- 10719043
OWN - NLM
STAT- MEDLINE
DCOM- 20000512
LR  - 20190513
IS  - 0267-8357 (Print)
IS  - 0267-8357 (Linking)
VI  - 15
IP  - 2
DP  - 2000 Mar
TI  - Isolation of DNA probes specific for rat chromosomal regions 19p, 19q and 4q and 
      their application for the analysis of diethylstilbestrol-induced aneuploidy in 
      binucleated rat fibroblasts.
PG  - 165-75
AB  - DNA probes specific for rat chromosomes 19p, 19q and 4q were isolated, 
      characterized and used for the detection and analysis of 
      diethylstilbestrol(DES)-induced aneuploidy. By denaturing and partially 
      reassociating total genomic DNA a new rat repetitive DNA family was isolated, 
      which was located on chromosome 19p21. Sequencing of a number of subclones from 
      cos76-1 and other clones of this so-called 76-family revealed that the repeat 
      units are interrupted with large areas of other (unique) DNA. Consequently, after 
      fluorescence in situ hybridization (FISH) the signals in interphase nuclei are 
      large and spread out. The other two probes, cos25 (chromosome 4q) and cos42-47 
      (chromosome 19q), were isolated by screening cosmid libraries with probes 
      isolated previously in our laboratory. The repeat unit of cos25 is a 2174 bp long 
      EcoRI unit that contains three Sau3A sites and is tandemly organized. Sequencing 
      of subclones of cos42-47 revealed that this probe was in fact the 5S RNA gene, 
      located on 19q12. In order to determine if these probes were suitable probes for 
      aneuploidy detection, two series of dual colour FISH with the combinations 
      cos25/cos76-1 (4q/19p) and cos42-47/cos76-1 (19q/19p) were carried out on slides 
      from an in vitro micronucleus assay with DES. With all three probes used, an 
      increase in binucleated cells with non-disjunction or chromosome loss was 
      observed in the DES-treated cultures. Scoring of additional micronucleated cells 
      on slides hybridized with the cos25/cos76-1 (4q/19p) probes revealed that the 
      hybridization signal of probe cos25 (4q) was over-represented in the micronuclei 
      of the control cultures. The simultaneous use of the 19q and 19p probes is a 
      particularly valuable tool for the detection of aneuploidy, since it allows 
      distinction between aneugenic and clastogenic events in binucleated cells. 
      Results of this analysis showed that apart from aneuploidy, DES also induced 
      structural chromosome aberrations, although to a lesser extent.
FAU - de Stoppelaar, J M
AU  - de Stoppelaar JM
AD  - Laboratory of Health Effects Research, National Institute of Public Health and 
      Environment, PO Box 1, 3720 BA Bilthoven, The Netherlands.
FAU - Faessen, P
AU  - Faessen P
FAU - Zwart, E
AU  - Zwart E
FAU - Hozeman, L
AU  - Hozeman L
FAU - Hodemaekers, H
AU  - Hodemaekers H
FAU - Mohn, G R
AU  - Mohn GR
FAU - Hoebee, B
AU  - Hoebee B
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Mutagenesis
JT  - Mutagenesis
JID - 8707812
RN  - 0 (Carcinogens)
RN  - 0 (DNA Probes)
RN  - 731DCA35BT (Diethylstilbestrol)
SB  - IM
MH  - *Aneuploidy
MH  - Animals
MH  - Bacteriophages
MH  - Base Sequence
MH  - Blotting, Southern
MH  - Carcinogens/*pharmacology
MH  - Cell Division/drug effects
MH  - Chromosomes/*genetics
MH  - Cosmids
MH  - *DNA Probes
MH  - Diethylstilbestrol/*pharmacology
MH  - Fibroblasts/*drug effects
MH  - Gene Library
MH  - In Situ Hybridization, Fluorescence
MH  - Male
MH  - Micronucleus Tests/methods
MH  - Models, Genetic
MH  - Molecular Sequence Data
MH  - Nondisjunction, Genetic
MH  - Rats
MH  - Rats, Sprague-Dawley
MH  - Rats, Wistar
EDAT- 2000/03/17 09:00
MHDA- 2000/05/20 09:00
CRDT- 2000/03/17 09:00
PHST- 2000/03/17 09:00 [pubmed]
PHST- 2000/05/20 09:00 [medline]
PHST- 2000/03/17 09:00 [entrez]
AID - 10.1093/mutage/15.2.165 [doi]
PST - ppublish
SO  - Mutagenesis. 2000 Mar;15(2):165-75. doi: 10.1093/mutage/15.2.165.