PMID- 8671728
OWN - NLM
STAT- MEDLINE
DCOM- 19960930
LR  - 20190512
IS  - 0267-8357 (Print)
IS  - 0267-8357 (Linking)
VI  - 11
IP  - 2
DP  - 1996 Mar
TI  - The centromere as a target for the induction of chromosome damage in resting and 
      proliferating mammalian cells: assessment of mitomycin C-induced genetic damage 
      at kinetochores and centromeres by a micronucleus test in mouse splenocytes.
PG  - 133-8
AB  - The cytokinesis-block micronucleus assay (MN) on murine splenocytes was used for 
      the estimation of chromosome damage in a resting cell population in vivo that can 
      be induced to proliferate in vitro. Mitomycin C at different doses (10(-8), 6 x 
      10(-8), 10(-7), 6 x 10(-7) and 10(-6)M) was used to induce cytogenetic damage in 
      resting and cycling splenocytes. Antikinetochore antibodies (CREST) and 
      two-colour fluorescence in situ hybridization (FISH) with minor and major 
      satellite DNA were applied. These approaches allowed the detailed 
      characterization of the mechanisms by which MN originates, since it was possible 
      to identify breaks induced in pericentric heterochromatic (resulting in MN 
      containing the major but not the minor satellite DNA) or detachment/disruption of 
      kinetochore (resulting in different frequencies of MN containing kinetochore or 
      both probes). Based on the evidence that resting and cycling mouse splenocytes 
      are characterized by different spatial distribution of centromeric regions, the 
      hypothesis was tested that the damage induced by mutagens at centromeres is 
      influenced by the phase of the cell cycle in which the cells are treated. Data 
      presented here show that resting and cycling splenocytes are both sensitive to 
      mitomycin C action, and indicate that this compound has an aneugenic potential, 
      besides its strong clastogenic activity. In particular, results obtained after 
      CREST and FISH characterization of MN differed when cells were treated during 
      proliferation, suggesting a disruption/detachment of kinetochores induced by 
      mitomycin C at this cell stage. Furthermore, under the same treatment condition 
      the proportion of MN containing the major satellite DNA only was greater than 
      expected on the basis of random breakage at this site. Treatment of resting cells 
      produced aneugenic damage, but without evidence of disruption/detachment of 
      kinetochores or preferential breakage at the centromere. These results indicate 
      that the amount and type of chromosome damage induced by mitomycin C in mouse 
      splenocytes differ in relation to the proliferative status of treated cells.
FAU - Renzi, L
AU  - Renzi L
AD  - Department of Biology, University of Padova, Italy.
FAU - Pacchierotti, F
AU  - Pacchierotti F
FAU - Russo, A
AU  - Russo A
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Mutagenesis
JT  - Mutagenesis
JID - 8707812
RN  - 11028-71-0 (Concanavalin A)
RN  - 50SG953SK6 (Mitomycin)
SB  - IM
MH  - Animals
MH  - Cell Cycle
MH  - Cell Division
MH  - Cells, Cultured
MH  - Centromere/*drug effects/genetics
MH  - Concanavalin A/pharmacology
MH  - DNA Damage
MH  - Immunoassay
MH  - In Situ Hybridization, Fluorescence
MH  - Kinetochores/*drug effects/immunology
MH  - Male
MH  - Mice
MH  - Mice, Inbred BALB C
MH  - Micronucleus Tests
MH  - Mitomycin/*toxicity
MH  - Spleen/cytology
EDAT- 1996/03/01 00:00
MHDA- 1996/03/01 00:01
CRDT- 1996/03/01 00:00
PHST- 1996/03/01 00:00 [pubmed]
PHST- 1996/03/01 00:01 [medline]
PHST- 1996/03/01 00:00 [entrez]
AID - 10.1093/mutage/11.2.133 [doi]
PST - ppublish
SO  - Mutagenesis. 1996 Mar;11(2):133-8. doi: 10.1093/mutage/11.2.133.