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Immunocytochemical Discrimination Between Early and Late S Phase: A New Approach to the Study of Gingival Epithelium Proliferation in Rats
Author(s) -
Breschi Lorenzo,
Mazzoni Giovanni,
Baratta Brunella,
Galanzi Angela,
Strocchi Paola,
Falconi Mirella,
Centurione Maria Antonietta,
Ferrari Carlo,
Rizzoli Riccardo
Publication year - 1998
Publication title -
journal of periodontology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.036
H-Index - 156
eISSN - 1943-3670
pISSN - 0022-3492
DOI - 10.1902/jop.1998.69.1.84
Subject(s) - bromodeoxyuridine , epithelium , cell growth , thymidine , biology , dna synthesis , cell division , microbiology and biotechnology , proliferation marker , chemistry , cell , dna , biochemistry , genetics
T he renewal of the free gingival margin epithelium in rats was studied evaluating 5‐bromodeoxyuridine (BrdU) incorporation in proliferating cells by means of an immunocytochemical method. We found a close correspondence between light and electron microscopy patterns of BrdU incorporation at a nuclear level. BrdU was localized in the inner interchromatin regions in cells starting DNA synthesis, while it was localized in the peripheral heterochromatin domains in cells terminating the S phase. This possibility of discriminating cells in early S phase from cells in late S is able to provide far more information as to the time at which a labeled cell starts proliferation than that obtainable with 3 H‐thymidine autoradiography. This, in turn, permits detection of cells that start proliferation in a wide period of time by means of a single BrdU administration. Rats treated at 7 a.m. demonstrated higher proliferation than rats treated at 7 p.m., supporting the existence of circadian variations in the epithelial renewal. Proliferative events take place by consecutive activation of at least three replication waves, producing clusters of labeled cells which could be observed in rats sacrificed at 10 a.m. In rats treated once with BrdU at 7 a.m., the clusters were localized in both the basal and suprabasal layer of the epithelium; in rats further injected with BrdU at the same time, the clusters increased in size, progressively extending throughout the epithelium. In this way, the renewal of the free gingival margin epithelium does not proceed randomly, but by consecutive activation of discrete units or clusters of basal cells, which then extend to the upper layers. This can be followed at a morphological level as a progression of labeled cells, which move from the basal layer to the epithelium surface in approximately 82–85 hours. J Periodontol 1998;69:84–91 .