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Nucleic Acids Research, 1992, Vol. 20, No. 23 6195-6200
© 1992

MOLECULAR BIOLOGY

New data concerning the functional organization of the mammalian cell nucleolus: detection of RNA and rRNA by in situ molecular immunocytochemistry

Marc Thiry

Laboratory of Cellular and Tissular Biology, University of Liège 20 rue de Pitteurs, B-4020 Liège, Belgium

Received October 6, 1992. Revised November 9, 1992. Accepted November 9, 1992.

We have investigated the fine spatial distribution of RNA and rRNA within the Ehrlich tumor cell nucleolus by in situ hybridization with a blotin-labeled probe and by two new strategies, the polyadenylate nucleotidyl transferase-immunogold technique and immunolabeling with anti-RNA antibodies. Besides the presence, as expected, of RNA and rRNA in the granular component and the dense fibrillar component, we show, for the first time, significant label over all the fibrillar centers of the nucleoli. When RNA and DNA were detected simultaneously on the same sections, only the fibrillar centers were positive for both. These results throw light on the controversial subject of the precise location of transcribing rRNA genes within the nucleolus. The fibrillar centers, and not the dense fibrillar component, should thus be the site of rRNA synthesis.


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