Human ribosomal RNA gene repeats are localized in the dense fibrillar component of nucleoli: Light and electron microscopic in situ hybridization in human Sertoli cells

1992 ◽  
Vol 198 (1) ◽  
pp. 135-143 ◽  
Author(s):  
F. Wachtler ◽  
C. Schöfer ◽  
W. Mosgöller ◽  
K. Weipoltshammer ◽  
H.G. Schwarzacher ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Ribosomal Rna ◽  
Sertoli Cells ◽  
Dense Fibrillar Component ◽  
Electron Microscopic ◽  
Ribosomal Rna Gene ◽  
Fibrillar Component

A study on nucleolar DNA: isolation of DNA from fibrillar components and ultrastructural localization of different DNA probes

1993 ◽  
Vol 104 (4) ◽  
pp. 1199-1205 ◽  
Author(s):  
P. Hozak ◽  
C. Schofer ◽  
J. Sylvester ◽  
F. Wachtler
Keyword(s):  
Ribosomal Rna ◽  
Sertoli Cells ◽  
Ultrastructural Localization ◽  
Ribosomal Rna Genes ◽  
Border Region ◽  
Dense Fibrillar Component ◽  
Mitotic Chromosomes ◽  
Electron Microscopic ◽  
Fibrillar Component ◽  
Rna Genes

The nature and localization of DNA contained in the fibrillar centres and the dense fibrillar component (the fibrillar complex) in the nucleoli, was studied in human LEP cells, Sertoli cells, spermatogonia A and in mitotic chromosomes of stimulated lymphocytes. A novel procedure for isolating the intact fibrillar complex from LEP cells was used; the complex contains DNA that hybridizes to secondary constrictions of mitotic chromosomes and to 28 S rDNA sequences, on Southern blots. Electron microscopic DNA-DNA in situ hybridization was performed, with (a) a probe prepared from DNA extracted from the fibrillar complex of LEP cells, (b) a probe for human total genomic DNA, and (c) a probe for the transcribed part of human rDNA. On the basis of the results obtained we conclude that the ribosomal RNA genes in human Sertoli cells and spermatogonia A are predominantly associated with the dense fibrillar component, including the border region between fibrillar centres and the dense fibrillar component. The ribosomal RNA genes are the main, if not exclusive, DNA type present in the fibrillar complex in the studied cell types.

scholarly journals Detection of the Ribosomal RNA Gene in Pear (Pyrus spp.) using Fluorescence in situ Hybridization

2010 ◽  
Vol 79 (4) ◽  
pp. 335-339 ◽  
Author(s):  
Masashi Yamamoto ◽  
Shingo Terakami ◽  
Toshiya Yamamoto ◽  
Norio Takada ◽  
Tatsuya Kubo ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Ribosomal Rna ◽  
Ribosomal Rna Gene

Cytochemical features common to nucleoli and cytoplasmic nucleoloids of Olea europaea meiocytes: detection of rRNA by in situ hybridization

1994 ◽  
Vol 107 (2) ◽  
pp. 621-629 ◽  
Author(s):  
J.D. Alche ◽  
M.C. Fernandez ◽  
M.I. Rodriguez-Garcia
Keyword(s):  
In Situ Hybridization ◽  
Olea Europaea ◽  
Electron Microscopic ◽  
Phosphorylated Proteins ◽  
Immunocytochemical Techniques ◽  
Fibrillar Component ◽  
Immunohistochemical Techniques ◽  
Microscopic Techniques ◽  
Olea Europaéa

We used light and electron microscopic techniques to study the composition of cytoplasmic nucleoloids during meiotic division in Olea europaea. Nucleoloids were found in two clearly distinguishable morphological varieties: one similar in morphology to the nucleolus, and composed mainly of dense fibrillar component, and another surrounded by many ribosome-like particles. Cytochemical and immunocytochemical techniques showed similar reactivities in nucleoloids and the nucleolus: both are ribonucleoproteic in nature, and possess argyrophillic, argentaffinic and highly phosphorylated proteins. Immunohistochemical techniques failed to detect DNA in either structure. In situ hybridization to a 18 S rRNA probe demonstrated the presence of ribosomal transcripts in both the nucleolus and nucleoloids. These similarities in morphology and composition may reflect similar functionalities.

Genome differentiation in Aegilops. 2. Physical mapping of 5S and 18S–26S ribosomal RNA gene families in diploid species

Genome ◽  
1996 ◽  
Vol 39 (6) ◽  
pp. 1150-1158 ◽  
Author(s):  
Ekatherina D. Badaeva ◽  
Bernd Friebe ◽  
Bikram S. Gill
Keyword(s):  
In Situ Hybridization ◽  
Ribosomal Rna ◽  
5S Rdna ◽  
Gene Families ◽  
Aegilops Species ◽  
Ribosomal Rna Gene ◽  
26S Rdna ◽  
Genome Differentiation ◽  
Rdna Loci

The distribution of the 5S and 18S–5.8S–26S (18S–26S) ribosomal RNA (rRNA) gene families on chromosomes of all diploid Aegilops species was studied by in situ hybridization with pTa71 (18S–26S rDNA) and pTa794 (5S rDNA) DNA clones. One major 18S–26S rDNA locus was found in the nucleolus organizer region (NOR) of each of the species Aegilops tauschii and Aegilops uniaristata and two loci were detected in the remaining species. In addition to major NORs, from one to nine minor loci were observed; their numbers and chromosomal locations were species-specific. Some minor loci were polymorphic, whereas others were conserved. One or two 5S rDNA loci were observed in the short arms of the chromosomes of groups 1 and 5 of all diploid Aegilops species except Ae. uniaristata, where one 5S rDNA site was located in the distal part of the long arm of chromosome 1N. The 5S rDNA loci were not associated with NORs; however, the relative positions of two ribosomal RNA gene families were diagnostic for chromosomes of homoeologous groups 1, 5, and 6. Implications of these results for establishing phylogenetic relationships of diploid Aegilops species and mechanisms of genome differentiation are discussed. Key words : wheat, Triticum, Aegilops, 5S rRNA, 18S–26S rRNA, in situ hybridization, evolution.

scholarly journals Ultrastructural distribution of DNA and RNA within the nucleolus of human Sertoli cells as seen by molecular immunocytochemistry

1993 ◽  
Vol 105 (1) ◽  
pp. 33-39 ◽  
Author(s):  
M. Thiry
Keyword(s):  
Sertoli Cells ◽  
Ultrastructural Level ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Dense Fibrillar Component ◽  
Granular Component ◽  
Dna And Rna ◽  
Nucleotidyl Transferase ◽  
Immunocytochemical Techniques ◽  
Fibrillar Component

The precise distribution of DNA and RNA within the human Sertoli cell nucleolus has been investigated, at the ultrastructural level, by cytochemical and molecular immunocytochemical techniques. In Sertoli cells, the nucleolar components show a typical spatial distribution. The fibrillar centres are not surrounded by a layer of dense fibrillar component, but come in contact only with strands of dense fibrillar component. These fibrillar parts of strands are the extensions of granular strands connected to a large granular mass. These strands delimit numerous nucleolar interstices in which chromatin fibres are clearly obvious. Using the in situ terminal deoxynucleotidyl transferase/immunogold procedure for detecting DNA, we find evident label exclusively over the chromatin fibres enclosed in the nucleolar interstices and over the fibrillar centres, and no significant label over the dense fibrillar component and granular component of the nucleolus. Furthermore, using the polyadenylate nucleotidyl transferase/immunogold procedure for detecting RNA, we show that label is deposited not only over the granular component and dense fibrillar component, as expected, but also quite obviously over the fibrillar centres. No label is seen over the interstices containing chromatin.

Detection of rRNA and phaseolin genes on polytene chromosomes of Phaseolus coccineus by fluorescence in situ hybridization after pepsin pretreatment

Genome ◽  
1994 ◽  
Vol 37 (6) ◽  
pp. 1018-1021 ◽  
Author(s):  
M. Nenno ◽  
K. Schumann ◽  
W. Nagl
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Ribosomal Rna ◽  
Storage Protein ◽  
Seed Storage Protein ◽  
Polytene Chromosomes ◽  
Seed Storage ◽  
Phaseolus Coccineus ◽  
Rna Genes

This is the first report of fluorescence in situ hybridization (FISH) on plant polytene chromosomes. Different protease pretreatments have been tested to improve fluorescence in situ hybridization FISH on polytene chromosomes of a plant, Phaseolus coccineus, with the aim to enable the detection of low-copy genes. The structural preservation of the chromosomes and the distinctness of the FISH signals were comparatively analysed with a probe for the ribosomal RNA genes after digestion with pepsin and trypsin. The pepsin pretreatment resulted in a general loosening of chromatin with good conservation of chromosome morphology and an increased number and density of signal points. The six nucleolus organizers exhibited significant differences in condensation. The pretreatment with pepsin enabled the detection of the low-copy genes encoding the seed storage protein phaseolin.Key words: plant, Leguminosae, ribosomal RNA genes, seed storage protein genes, protease.

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