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Immunoelectron microscopic study of nucleolar DNA during mitosis in Ehrlich tumor cells.
Thiry, Marc; Scheer, U.; Goessens, Guy
1988In European Journal of Cell Biology, 47 (2), p. 346-57
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Keywords :
Animals; Antibodies, Monoclonal/diagnostic use/immunology; Bromodeoxyuridine/immunology; Carcinoma, Ehrlich Tumor/pathology/ultrastructure; DNA, Neoplasm/analysis/ultrastructure; Dactinomycin/pharmacology; Interphase; Metaphase; Mice; Mice, Inbred Strains; Microscopy, Electron/methods; Mitosis; Prophase; Telophase
Abstract :
[en] In order to investigate the DNA localization within Ehrlich tumor cell nucleoli during mitosis, two recent immunocytochemical methods using either an anti-DNA or an anti-bromodeoxyuridine (BrdU) monoclonal antibody have been applied. In both cases, the immunogold labeling has been performed on ultrathin sections of cells embedded either in Lowicryl K4M or in Epon, respectively. Identical results are observed with both immunocytochemical approaches. In the interphase nucleolus, besides the labeling of the perinucleolar chromatin shell and of its intranucleolar invaginations which penetrate into the nucleolar body and often terminate at the fibrillar centers, a few gold particles are also preferentially found towards the peripheral region of the fibrillar centers. In contrast, the dense fibrillar component and the granular component are never labeled. During mitosis, the fibrillar centers persist at the chromosomal nucleolus organizing regions (NOR's) and can be selectively stained by the silver method. However, these metaphase fibrillar centers are no longer decorated by the DNA- or BrdU antibodies. These results indicate that until the end of prophase, rRNA genes are present inside the fibrillar center material, disappear during metaphase and reappear in reconstituting nucleoli during telophase. Thus, fibrillar centers appear to represent structures sui generis, which are populated by rRNA genes only when the nucleolus is functionally active. In segregated nucleoli after actinomycin D treatment, the DNA labeling is exclusively restricted to the perinucleolar chromatin blocks. These findings also suggest that the DNA content of the fibrillar center material varies according to the rRNA transcription level of the cells. The results are discussed in the light of the present knowledge of the functional organization of the nucleolus.
Disciplines :
Anatomy (cytology, histology, embryology...) & physiology
Author, co-author :
Thiry, Marc  ;  Université de Liège - ULiège > Département des sciences de la vie > Biologie cellulaire
Scheer, U.
Goessens, Guy ;  Université de Liège - ULiège > Services généraux (Faculté des sciences) > Relations académiques et scientifiques (Sciences)
Language :
English
Title :
Immunoelectron microscopic study of nucleolar DNA during mitosis in Ehrlich tumor cells.
Publication date :
1988
Journal title :
European Journal of Cell Biology
ISSN :
0171-9335
eISSN :
1618-1298
Publisher :
Wissenschaftliche Verlagsgesellschaft, Stuttgart, Germany
Volume :
47
Issue :
2
Pages :
346-57
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 24 November 2009

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