High-resolution detection of newly synthesized DNA by anti-bromodeoxyuridine antibodies identifies specific chromatin domains

G. Mazzotti, R. Rizzoli, A. Galanzi, S. Papa, M. Vitale, M. Falconi, L. M. Neri, N. Zini, N. M. Maraldi

Research output: Contribution to journalArticlepeer-review

Abstract

We analyzed the incorporation of bromodeoxyuridine (BrdUrd) into DNA in exponentially growing murine erythroleukemia cells (FLC-745), using fluorescent anti-BrdUrd antibodies with light microscopy and flow cytometry. The fine localization of the DNA replicating sites was investigated at the ultrastructural level by using a second antibody conjugated with colloidal gold. The latter approach, which does not require acidic denaturation of the DNA, enables preservation of good morphology and obtains a better resolution power than that of electron microscopic autoradiography, the percentage of labeled cells obtained with the two techniques being comparable. After short BrdUrd pulses, characteristic distribution of the labeling can be identified in the heterochromatin, in interchromatin domains, or at the boundary between the dispersed and the condensed chromatin. Similar patterns are also observable in the nuclear structures which condense after acid denaturation, suggesting that DNA replication takes place at fixed sites associated with the nuclear matrix.

Original languageEnglish
Pages (from-to)13-22
Number of pages10
JournalJournal of Histochemistry and Cytochemistry
Volume38
Issue number1
Publication statusPublished - 1990

Keywords

  • bromodeoxyuridine
  • cell cycle
  • DNA detection
  • DNA synthesis
  • electron microscopy
  • flow cytometry
  • immunocytochemistry

ASJC Scopus subject areas

  • Cell Biology
  • Anatomy

Fingerprint Dive into the research topics of 'High-resolution detection of newly synthesized DNA by anti-bromodeoxyuridine antibodies identifies specific chromatin domains'. Together they form a unique fingerprint.

Cite this