Reference : Isolation of nucleoli from ELT cells: a quick new method that preserves morphological...
Scientific journals : Article
Life sciences : Anatomy (cytology, histology, embryology...) & physiology
Isolation of nucleoli from ELT cells: a quick new method that preserves morphological integrity and high transcriptional activity.
Vandelaer, M. [> > > >]
Thiry, Marc mailto [Université de Liège - ULiège > Département des sciences de la vie > Biologie cellulaire >]
Goessens, Guy mailto [Université de Liège - ULiège > Services généraux (Faculté des sciences) > Relations académiques et scientifiques (Sciences) >]
Experimental Cell Research
Academic Press
Yes (verified by ORBi)
[en] Animals ; Buffers ; Carcinoma, Ehrlich Tumor/genetics/metabolism/ultrastructure ; Cell Nucleolus/metabolism/ultrastructure ; Cell Separation/methods ; Mice ; Microscopy, Electron ; Nucleolus Organizer Region/ultrastructure ; RNA, Neoplasm/biosynthesis ; RNA, Ribosomal/biosynthesis ; Silver ; Staining and Labeling/methods ; Transcription, Genetic ; Tumor Cells, Cultured
[en] We have developed a quick new method for isolating nucleoli which, unlike the methods in current use, preserves the nucleolar ultrastructure. Until now, the isolation process has generally been assumed to empty one of the three major compartments of the nucleolus, the fibrillar center, of its content. We have used the AgNOR staining and in vitro transcription assay to test the degree of structural and functional preservation of the isolated nucleoli. Our results demonstrate the value of our procedure as a reliable tool for biochemical and ultrastructural studies on the nucleolus. Moreover, these proprieties prompt us to investigate the rRNA synthesis, using a nonisotopic approach, within morphologically intact isolated nucleoli. Thus, we show that newly synthesized rRNA transcripts are located not only in the dense fibrillar component, but also indubitably in the fibrillar center.

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