Combination of electron microscopic in situ hybridization and anti-DNA antibody labelling reveals a peculiar arrangement of ribosomal DNA in the fibrillar centres of the plant cell nucleolus.
The fibrillar centres (FCs) in the nucleoli of Allium cepa usually contained compact dense chromatin, which was always surrounded with light fibrous material (LFM). Distribution of 18S ribosomal DNA (rDNA) in the FCs was examined by in situ hybridization at the light and electron microscopic levels and the results were compared with those obtained by immunogold labelling with anti-DNA antibodies. Anti-DNA antibodies heavily labelled the dense chromatin of the FCs but scarcely labelled the LFM. However, electron microscopic in situ hybridization using the 18S rDNA probe showed that the label in the dense chromatin was extremely weak compared with that obtained by the anti-DNA antibody labelling: the specific label with anti-DNA antibodies of the dense chromatin was about 15 times as much as that of the LFM, whereas the specific label with in situ hybridization in the dense chromatin was only about 1.7 times higher than in the LFM. These results suggest that the rDNA encoding rRNA is preferentially released from the dense chromatin and that non-transcribed intergenic spacers remain in the dense chromatin as the anchoring sites of rDNA.
Yano
Hiroyuki
H
Department of Biology and Earth Sciences, Faculty of Science, Ehime University, Matsuyama, Japan.
Sato
Seiichi
S
D000906:Antibodies / Q000276:immunology; D002466:Cell Nucleolus / Q000648:ultrastructure*; D002843:Chromatin / Q000648:ultrastructure*; D018744:DNA, Plant / Q000276:immunology; D004275:DNA, Ribosomal / Q000648:ultrastructure*; D017403:In Situ Hybridization; D008854:Microscopy, Electron; D019697:Onions / Q000235:genetics / Q000648:ultrastructure*; D012337:RNA, Ribosomal, 18S / Q000737:chemistry
D000906:Antibodies; D002843:Chromatin; D018744:DNA, Plant; D004275:DNA, Ribosomal; D012337:RNA, Ribosomal, 18S