In situ fluorescence visualization of bromouridine incorporated into newly transcribed nucleolar RNA
Language English Country Germany Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
- MeSH
- Bromouracil analogs & derivatives MeSH
- Cell Nucleolus metabolism MeSH
- Fibroblasts cytology metabolism MeSH
- Fluorescent Antibody Technique, Indirect MeSH
- Transcription, Genetic * MeSH
- HeLa Cells cytology metabolism MeSH
- In Situ Hybridization, Fluorescence MeSH
- Cells, Cultured MeSH
- Kidney cytology metabolism MeSH
- Humans MeSH
- Nucleolus Organizer Region metabolism MeSH
- Dogs MeSH
- RNA, Ribosomal biosynthesis MeSH
- RNA Polymerase I metabolism MeSH
- Uridine analogs & derivatives metabolism MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Dogs MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- 5-bromouridine MeSH Browser
- Bromouracil MeSH
- RNA, Ribosomal MeSH
- RNA Polymerase I MeSH
- Uridine MeSH
Bromouridine-triphosphate is commonly used for in situ immunocytochemical labeling of newly synthesized RNA in living cells. While extranucleolar transcripts do not require special conditions for visualization, special treatment prior to fixation (e.g. incubation with alpha-amanitine) is necessary for immunofluorescence detection of bromouridine-labeled nucleolar RNA in previous studies. We show in the present investigation that bromouridine-triphosphate is efficiently used by both extranucleolar and nucleolar RNA polymerases in living cultured cells. The failure to detect incorporated bromouridine within nucleoli is entirely due to improper treatment of cells after bromouridine incorporation. When methanol/acetone fixation is used, fluorescence signals within nucleoli can be routinely found.
References provided by Crossref.org
NORs and their transcription competence during the cell cycle
