PMID- 9269338
OWN - NLM
STAT- MEDLINE
DCOM- 19970910
LR  - 20190702
IS  - 0027-5107 (Print)
IS  - 0027-5107 (Linking)
VI  - 392
IP  - 1-2
DP  - 1997 Aug 1
TI  - Analysis of DES-induced micronuclei in binucleated rat fibroblasts: comparison 
      between FISH with a rat satellite I probe and immunocytochemical staining with 
      CREST serum.
PG  - 139-49
AB  - The usefulness of fluorescence in situ hybridization (FISH) with rat satellite I 
      DNA was compared with immunocytochemical staining with CREST serum for the 
      analysis of the content of micronuclei from primary rat fibroblasts. We analyzed 
      micronuclei induced in vitro by the aneugenic compound diethylstilbestrol (DES) 
      or the clastogenic compound mitomycin C (MMC). Since a centromeric probe was not 
      available for the rat, we isolated rat satellite I DNA by PCR with primers 
      designed on the basis of the known rat satellite I DNA sequence. The PCR products 
      obtained as well as the cloned PCR products showed hybridization to the 
      centromeric regions of a large number of chromosomes, but not of chromosome 1, 
      19, 20, X and Y. Clone 18-5 was further analyzed and was shown to contain at 
      least 4 repeats of the rat satellite I family. This probe, which hybridizes in 
      the centromeric region of 34 of the 42 chromosomes, was used throughout the study 
      as a probe for the FISH analysis of the micronuclei. For the immunocytochemical 
      staining, the commonly used commercial anti-centromeric antibodies could not be 
      used because of the weakness of the fluorescent signals given. Consequently, 
      CREST serum of a single patient was used, which showed bright and distinct 
      signals on the kinetochores of each chromosome. After treatment of the cells with 
      the aneugen DES an increase in centromere (FISH) and kinetochore (CREST) positive 
      micronuclei was found, whereas after treatment with the clastogen MMC, the 
      percentage of centromere-positive micronuclei was similar to that observed in 
      controls. Analysis of a large number of DES-induced micronuclei showed that the 
      immunocytochemical method is equally as or slightly less sensitive for the 
      detection of chromosomes in micronuclei and we therefore recommend FISH with 
      probe 18-5 for the detection of chromosome loss in rat cells.
FAU - de Stoppelaar, J M
AU  - de Stoppelaar JM
AD  - Department of Carcinogenesis, Mutagenesis and Genetics, National Institute of 
      Public Health and Environment, Bilthoven, Netherlands. JM.de.Stoppelaar@rivm.nl
FAU - de Roos, B
AU  - de Roos B
FAU - Mohn, G R
AU  - Mohn GR
FAU - Hoebee, B
AU  - Hoebee B
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Netherlands
TA  - Mutat Res
JT  - Mutation research
JID - 0400763
RN  - 0 (Autoantibodies)
RN  - 0 (DNA Probes)
RN  - 0 (DNA, Satellite)
RN  - 0 (Mutagens)
RN  - 50SG953SK6 (Mitomycin)
RN  - 731DCA35BT (Diethylstilbestrol)
SB  - IM
MH  - Animals
MH  - Autoantibodies
MH  - CREST Syndrome/immunology
MH  - Cells, Cultured
MH  - Centromere/genetics
MH  - *DNA Probes
MH  - DNA, Satellite
MH  - Diethylstilbestrol
MH  - Fibroblasts
MH  - Fluorescent Antibody Technique, Indirect/*methods
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Male
MH  - Micronucleus Tests/*methods
MH  - Mitomycin
MH  - Mutagens
MH  - Rats
MH  - Rats, Wistar
EDAT- 1997/08/01 00:00
MHDA- 1997/08/01 00:01
CRDT- 1997/08/01 00:00
PHST- 1997/08/01 00:00 [pubmed]
PHST- 1997/08/01 00:01 [medline]
PHST- 1997/08/01 00:00 [entrez]
AID - S0165-1218(97)00052-9 [pii]
AID - 10.1016/s0165-1218(97)00052-9 [doi]
PST - ppublish
SO  - Mutat Res. 1997 Aug 1;392(1-2):139-49. doi: 10.1016/s0165-1218(97)00052-9.