The 3-D substructure of RNA in nascent RNP granules: A novel application of osmium ammine-B staining and electron spectroscopic imaging

1992 ◽  
Vol 50 (1) ◽  
pp. 504-505
Author(s):  
Ada L. Olins ◽  
Donald E. Olins ◽  
Manesh B. Shah ◽  
Henri A. Levy ◽  
David P. Bazett-Jonest
Keyword(s):  
Salivary Gland ◽  
Polytene Chromosomes ◽  
Spectroscopic Imaging ◽  
Secretory Proteins ◽  
Chironomus Tentans ◽  
Balbiani Ring ◽  
Electron Spectroscopic Imaging ◽  
Nuclear Pores ◽  
Rnp Granules

RNA has a particulate substructure when visualized in situ with the nucleic acid specific stain osmium ammine-B (OA-B). In this study energy spectroscopic imaging (ESI) was used to enhance the contrast and collect the data for tomographic reconstructions.The Balbiani ring (BR) in the salivary gland polytene chromosomes of Chironomus tentans larvae furnishes a well known model for the structure of nascent m-RNA. This gland produces copious amounts of silk-like secretory proteins which are very large (106 daltons). The site of transcription, the BR, is easily recognized in the EM by its characteristic “puff” structure and electron-dense granular transcripts. Mature BR granules are 45-50 nm in diameter and can be easily observed within the nucleus and passing through nuclear pores.

Microinjection of anti-topoisomerase I immunoglobulin G into nuclei of Chironomus tentans salivary gland cells leads to blockage of transcription elongation

1987 ◽  
Vol 7 (12) ◽  
pp. 4308-4316
Author(s):  
E Egyházi ◽  
E Durban
Keyword(s):  
Salivary Gland ◽  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Immunoglobulin G ◽  
Topoisomerase I ◽  
Chironomus Tentans ◽  
Balbiani Ring ◽  
Gland Cells ◽  
Salivary Gland Cells ◽  
Transcriptional Process

Purified anti-topoisomerase I immunoglobulin G (IgG) was microinjected into nuclei of Chironomus tentans salivary gland cells, and the effect on DNA transcription was investigated. Synthesis of nucleolar preribosomal 38S RNA by RNA polymerase I and of chromosomal Balbiani ring RNA by RNA polymerase II was inhibited by about 80%. The inhibitory action of anti-topoisomerase I IgG could be reversed by the addition of exogenous topoisomerase I. Anti-topoisomerase I IgG had less effect on RNA polymerase II-promoted activity of other less efficiently transcribing heterogeneous nuclear RNA genes. The pattern of inhibition of growing nascent Balbiani ring chains indicated that the transcriptional process was interrupted at the level of chain elongation. The highly decondensed state of active Balbiani ring chromatin, however, remained unaffected after injection of topoisomerase I antibodies. These data are consistent with the interpretation that topoisomerase I is an essential component in the transcriptional process but not in the maintenance of the decondensed state of active chromatin.

The genome of the olive fruit fly Bactrocera oleae: localization of molecular markers by in situ hybridization to the salivary gland polytene chromosomes

Genome ◽  
1999 ◽  
Vol 42 (4) ◽  
pp. 744-751 ◽  
Author(s):  
Anna Zambetaki ◽  
Antigone Zacharopoulou ◽  
Zacharias G. Scouras ◽  
Penelope Mavragani-Tsipidou
Keyword(s):  
Salivary Gland ◽  
Molecular Markers ◽  
In Situ Hybridization ◽  
Polytene Chromosomes ◽  
Fruit Fly ◽  
Bactrocera Oleae ◽  
Olive Fruit Fly ◽  
Olive Fruit

scholarly journals UNDER-REPLICATION OF RIBOSOMAL CISTRONS IN POLYTENE CHROMOSOMES OF RHYNCHOSCIARA

1974 ◽  
Vol 62 (1) ◽  
pp. 215-222 ◽  
Author(s):  
A. G. Gambarini ◽  
F. J. S. Lara
Keyword(s):  
Salivary Gland ◽  
In Situ Hybridization ◽  
Xenopus Laevis ◽  
Related Species ◽  
Polytene Chromosomes ◽  
Chromosome X ◽  
Heterologous Hybridization ◽  
Ribosomal Cistrons ◽  
Rhynchosciara Americana

DNA preparations obtained from several tissues of Rhynchosciara americana and two related species, R. milleri and R. papaveroi, were hybridized to R. americana rRNA. The percentage of hybridization was found to be higher in tissues with low polyteny than in tissues with high polyteny, suggesting a relationship between the amount of rDNA and the tissue polyteny. This could be explained by under-replication of ribosomal cistrons in polytene cells, such as those from the salivary gland. Only slight tissue-dependent changes in the percentages of hybridization can be observed in heterologous hybridization using Xenopus laevis rRNA. The possibility that these experiments could not detect differences in the amount of ribosomal cistrons among tissues is discussed. The female:male ratio for the percentages of hybridization in the salivary gland of R. americana agrees with the results obtained by in situ hybridization experiments (16, 17) which have shown that the rRNA cistrons are distributed among chromosomes other than chromosome X.

The genome of the Mediterranean fruitflyceratitis capitata: Localization of molecular markers by in situ hybridization to salivary gland polytene chromosomes

Chromosoma ◽  
1992 ◽  
Vol 101 (7) ◽  
pp. 448-455 ◽  
Author(s):  
A. Zacharopoulou ◽  
M. Frisardi ◽  
C. Savakis ◽  
A. S. Robinson ◽  
P. Tolias ◽  
...  
Keyword(s):  
Salivary Gland ◽  
Molecular Markers ◽  
In Situ Hybridization ◽  
Polytene Chromosomes ◽  
The Mediterranean

Electron spectroscopic imaging of the morphology of in situ polyethylene ‘shish kebabs’

1992 ◽  
Vol 166 (3) ◽  
pp. 317-328 ◽  
Author(s):  
D. M. Huong ◽  
M. Drechsler ◽  
M. Möller ◽  
H.-J. Cantow
Keyword(s):  
Spectroscopic Imaging ◽  
Electron Spectroscopic Imaging

The Balbiani ring and the polytene chromosomes of Drosophila bicornuta

Genome ◽  
1992 ◽  
Vol 35 (1) ◽  
pp. 64-67 ◽  
Author(s):  
P. Mavragani-Tsipidou ◽  
Z. G. Scouras ◽  
A. Natsiou-Voziki
Keyword(s):  
Salivary Gland ◽  
Larval Development ◽  
Banding Pattern ◽  
Polytene Chromosomes ◽  
Balbiani Ring

A study of the BR1 and of the most prominent puffs during larval development and after in vitro ecdysterone treatment, as well as of the banding pattern and inverted tandem chromosomal duplications of the salivary gland chromosomes of Drosophila bicornuta, is presented in this report. These data are compared and discussed with those of D. auraria and D. serrata, two other montium species.Key words: Drosophila, Balbiani ring, duplications, ecdysterone.

scholarly journals Balbiani ring hnRNP substructure visualized by selective staining and electron spectroscopic imaging.

1992 ◽  
Vol 117 (3) ◽  
pp. 483-491 ◽  
Author(s):  
A L Olins ◽  
D E Olins ◽  
D P Bazett-Jones
Keyword(s):  
Nucleic Acid ◽  
Polytene Chromosomes ◽  
Particle Diameter ◽  
Balbiani Ring ◽  
Electron Spectroscopic Imaging ◽  
Selective Staining ◽  
Elemental Imaging ◽  
Mature Transcript ◽  
Balbiani Rings ◽  
Relationship Of

The Balbiani Rings (BR) in the polytene chromosomes of Chironomus salivary glands are intense sites of transcription. The nascent RNPs fold during transcription into 40-50-nm granules, containing in the mature transcript approximately 37-kb RNA. Using a new nucleic acid specific stain, osmium ammine B on Lowicryl sections, in combination with electron energy filtered imaging of sections containing BR granules, we demonstrate a RNA-rich particulate substructure (10-nm particle diameter; 10-12 particles per BR granule). Elemental imaging supports that these particles are enriched in phosphorus. The possible relationship of these RNA-rich particles to ribonucleosomes is discussed, as well as models for their arrangement in the mature BR granules.

scholarly journals Microinjection of anti-topoisomerase I immunoglobulin G into nuclei of Chironomus tentans salivary gland cells leads to blockage of transcription elongation.

1987 ◽  
Vol 7 (12) ◽  
pp. 4308-4316 ◽  
Author(s):  
E Egyházi ◽  
E Durban
Keyword(s):  
Salivary Gland ◽  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Immunoglobulin G ◽  
Topoisomerase I ◽  
Chironomus Tentans ◽  
Balbiani Ring ◽  
Gland Cells ◽  
Salivary Gland Cells ◽  
Transcriptional Process

Purified anti-topoisomerase I immunoglobulin G (IgG) was microinjected into nuclei of Chironomus tentans salivary gland cells, and the effect on DNA transcription was investigated. Synthesis of nucleolar preribosomal 38S RNA by RNA polymerase I and of chromosomal Balbiani ring RNA by RNA polymerase II was inhibited by about 80%. The inhibitory action of anti-topoisomerase I IgG could be reversed by the addition of exogenous topoisomerase I. Anti-topoisomerase I IgG had less effect on RNA polymerase II-promoted activity of other less efficiently transcribing heterogeneous nuclear RNA genes. The pattern of inhibition of growing nascent Balbiani ring chains indicated that the transcriptional process was interrupted at the level of chain elongation. The highly decondensed state of active Balbiani ring chromatin, however, remained unaffected after injection of topoisomerase I antibodies. These data are consistent with the interpretation that topoisomerase I is an essential component in the transcriptional process but not in the maintenance of the decondensed state of active chromatin.

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