Human Genetics

, Volume 94, Issue 6, pp 684–692 | Cite as

New sites of methylcytosine-rich DNA detected on metaphase chromosomes

  • Agnès Barbin
  • Claire Montpellier
  • Nadja Kokalj-Vokac
  • Anne Gibaud
  • Alain Niveleau
  • Bernard Malfoy
  • Bernard Dutrillaux
  • Claire A. Bourgeois
Original Investigation


In situ immunofluorescence detection of antibodies against 5-methylcytosine on metaphase chromosomes prepared by a new procedure allows the display of new 5-methylcytosine-rich sites as compared to previously published methods. In short-term culture lymphocytes, the immunofluorescent signals give a recurrent pattern in which four types of binding sites can be distinguished. Type I sites are the secondary constrictions and a few juxtacentromeric regions, type II sites correspond to T-bands. Both types I and II sites emit a strong fluorescence. Type III sites form an R-band pattern and emit a weaker fluorescence. Type IV sites are the short arms of acrocentrics, they emit strong but polymorphic signals. The results obtained from control experiments suggest that the pattern observed is rather the expression of an uneven distribution of 5-methylcytosine-rich sites than a consequence of the various treatments used. In a lymphoblastoid cell line known to have a reduced 5-methylcytosine content, it was possible to demonstrate a heterogeneous hypomethylation among chromosome structures, principally involving type I sites. The method opens the possibility of studying in situ on chromosomes, regional variations of methylation in pathological conditions.


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Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • Agnès Barbin
    • 1
  • Claire Montpellier
    • 2
  • Nadja Kokalj-Vokac
    • 3
  • Anne Gibaud
    • 2
  • Alain Niveleau
    • 4
  • Bernard Malfoy
    • 2
  • Bernard Dutrillaux
    • 2
  • Claire A. Bourgeois
    • 2
  1. 1.CNRS URA 147, Institut Gustave RoussyVillejuif CedexFrance
  2. 2.CNRS URA 620, Institut CurieParis CedexFrance
  3. 3.Cytogenetic LaboratoryMariborSlovenie
  4. 4.CNRS URA 1459. Institut PasteurLyon CedexFrance

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