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Revealing the unseen: the organizer region of the nucleolus

Marco Biggiogera^1,2,*, Manuela Malatesta^1,3,*, Sousan Abolhassani-Dadras¹, François Amalric⁴, Lawrence I. Rothblum⁵ and Stanislav Fakan^1,

¹ Centre of Electron Microscopy, University of Lausanne, 1005 Lausanne, Switzerland
² Dipartimento di Biologia Animale, Laboratorio di Istologia, and Centro di Studio per l’Istochimica del CNR, University of Pavia, 27100 Pavia, Italy
³ Istituto di Istologia ed Analisi di Laboratorio, University of Urbino, 61029 Urbino, Italy
⁴ Institut de Pharmacologie et de Biologie Structurale, CNRS, 31077 Toulouse, France
⁵ Weis Center for Research, Geisinger Clinic, Danville, PA 17822, USA
^* These authors contributed equally to this work

Author for correspondence (e-mail: sfakan{at}cme.unil.ch)

Accepted May 22, 2001

We carried out a high-resolution ultrastructural analysis of the nucleolus in mouse P815 cells by combining specific DNA and RNA staining, anti-fibrillarin immunolabeling, contrast enhancement by energy filtering TEM and phosphorus mapping by ESI to visualize nucleic acids. We demonstrated that specifically contrasted DNA, fibrillarin and phosphorus overlap within the nucleolar dense fibrillar component. Moreover, we describe a ‘DNA cloud’ consisting of an inner core of DNA fibers (fibrillar center) and a periphery made of extremely thin fibrils overlapping the anti-fibrillarin immunolabeling (dense fibrillar component). This highly sensitive approach has allowed us to demonstrate, for the first time, the exact distribution of DNA within the decondensed interphase counterpart of the NOR, which includes both the fibrillar center and the dense fibrillar component.

Key words: Nucleolus Organizer Region, Ultrastructure, Electron Spectrosopic Imaging, Immunocytochemistry

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