Visualization of transcription sites at the electron microscope.
In order to localize at EM level the sites of transcription of both pre-mRNA and pre-rRNA, we have detected the DNA/RNA hybrid molecules and m3Gcapped structures by means of specific antibodies after short bromo-uridine (BrU) incorporation. In addition, the sections have been stained by a selective RNA stain, terbium citrate. Our data indicate that perichromatin fibrils incorporate BrU and are labeled by the anti-hybrid probe; this supports the idea that they are the pre-mRNA transcription sites. On the contrary, interchromatin granules do not incorporate BrU after short pulses and are not labeled by the anti-hybrid probe. Concerning the nucleolus, anti-hybrid and anti-BrdU antibodies colocalize only on the dense fibrillar component, suggesting that this is the site of rRNA transcription. Interestingly, the dense fibrillar component and the granular component, after specific RNA staining, show remarkable structural similarities, both containing fibrogranular RNA structures.
D000818:Animals; D000906:Antibodies / Q000276:immunology; D001973:Bromodeoxyuridine; D001976:Bromouracil / Q000031:analogs & derivatives; D002478:Cells, Cultured; D004247:DNA / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D005347:Fibroblasts; D006801:Humans; D008854:Microscopy, Electron; D012313:RNA / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000648:ultrastructure*; D051381:Rats; D014158:Transcription, Genetic; D014529:Uridine / Q000031:analogs & derivatives* / Q000378:metabolism*
D000906:Antibodies; D001976:Bromouracil; D012313:RNA; D004247:DNA; D001973:Bromodeoxyuridine; C006824:5-bromouridine; D014529:Uridine