PMID- 7507215
OWN - NLM
STAT- MEDLINE
DCOM- 19940224
LR  - 20220311
IS  - 0027-5107 (Print)
IS  - 0027-5107 (Linking)
VI  - 312
IP  - 1
DP  - 1994 Feb
TI  - Measurement and characterization of micronuclei in exfoliated human cells by 
      fluorescence in situ hybridization with a centromeric probe.
PG  - 39-50
AB  - The micronucleus (MN) assay in human exfoliated cells has been widely used to 
      detect the genotoxic effects of environmental mutagens, infectious agents and 
      hereditary diseases. Substantial variability characterizes the MN frequencies 
      reported by different research groups. One reason for this may be the restricted 
      resolution power of the Feulgen-Fast-Green staining that is routinely used. Here 
      we describe a new version of the MN assay that employs fluorescent propidium 
      iodide staining along with fluorescence in situ hybridization (FISH) with a 
      centromeric probe. Buccal and urothelial cells were collected from 5 healthy 
      unexposed female volunteers and 55,000 cells analyzed for MN frequency and 
      abnormal nuclear events. The Feulgen-Fast-Green and the new fluorescent staining 
      produced very similar results. The frequency of MN in buccal cells was 0.145 +/- 
      0.118% and in urothelial cells 0.083 +/- 0.074%. No correlation was found between 
      the frequencies of MN in the two types of exfoliated cells. FISH with a 
      centromeric probe allowed MN containing whole chromosomes with a centromere to be 
      differentiated from those containing only acentric fragments. The former appear 
      as a result of chromosome lagging in mitosis, while those without a centromere 
      are due to chromosome breakage. In urothelial cells 43% of MN were 
      centromere-negative and in buccal cells -44%. Fluorescent staining provided more 
      accurate scoring of degenerative cells than standard Feulgen-Fast-Green staining. 
      The combined frequency of pycnotic cells, "broken eggs" and cells with fragmented 
      nuclei did not exceed 2%, while that of karyorrhexis and karyolysis together was 
      as high as 21%. Significant interindividual variability was found in the 
      frequency of cells with karyolysis and karyorrhexis. Thus, the new version of 
      micronucleus assay allows for MN to be scored more precisely, the mechanism of MN 
      formation to be determined and abnormal nuclear events to be readily identified 
      in exfoliated human cells. It is therefore ideal for studying genotoxicity in 
      human populations using exfoliated cells from the mouth, bladder and nose.
FAU - Titenko-Holland, N
AU  - Titenko-Holland N
AD  - Center for Occupational and Environmental Health, School of Public Health, 
      University of California at Berkeley 94720.
FAU - Moore, L E
AU  - Moore LE
FAU - Smith, M T
AU  - Smith MT
LA  - eng
GR  - P30 ES01896/ES/NIEHS NIH HHS/United States
GR  - P42 ES04705/ES/NIEHS NIH HHS/United States
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - Netherlands
TA  - Mutat Res
JT  - Mutation research
JID - 0400763
RN  - 0 (Coloring Agents)
RN  - 0 (DNA Probes)
RN  - 0 (Feulgen stain)
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Rosaniline Dyes)
RN  - 36015-30-2 (Propidium)
RN  - 9J3VQ0Y6BV (Fast Green)
SB  - IM
MH  - Adult
MH  - Analysis of Variance
MH  - Cell Nucleus/pathology
MH  - Centromere/*ultrastructure
MH  - Coloring Agents
MH  - DNA Probes
MH  - Epithelial Cells
MH  - Female
MH  - Fluorescent Dyes
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Karyometry/methods
MH  - Micronucleus Tests/*methods
MH  - Mouth Mucosa/cytology
MH  - Propidium
MH  - Reproducibility of Results
MH  - Rosaniline Dyes
MH  - Staining and Labeling/*methods
MH  - Urinary Bladder/cytology
EDAT- 1994/02/01 00:00
MHDA- 1994/02/01 00:01
CRDT- 1994/02/01 00:00
PHST- 1994/02/01 00:00 [pubmed]
PHST- 1994/02/01 00:01 [medline]
PHST- 1994/02/01 00:00 [entrez]
AID - 0165-1161(94)90007-8 [pii]
AID - 10.1016/0165-1161(94)90007-8 [doi]
PST - ppublish
SO  - Mutat Res. 1994 Feb;312(1):39-50. doi: 10.1016/0165-1161(94)90007-8.