Abstract
The relationship between the Ag-NOR (silverstained Nucleolar Organizer Region) proteins and the functional-structural organization of the nucleolar ribosomal chromatin was studied in regenerating and cortisol-stimulated rat hepatocytes. Statistical analysis of Ag-NOR proteins, carried out with an automated image analyzer, indicated that in regenerating rat hepatocytes the quantity of Ag-NOR proteins mainly increased between the 4th and 12th h of regeneration, reaching a level twice that of resting hepatocytes. Also the synthesis of pre-ribosomal RNA (pre-rRNA) was stimulated after the 4th of regeneration. Cycloheximide administered to rats at a dose of 0.025 mg/100 g body weight (bw) prevented any increase in Ag-NOR proteins but did not hinder the stimulation of pre-rRNA snythesis. In 8 h cortisol-stimulated hepatocytes no significant change in amount of Ag-NOR protein was observed whereas pre-rRNA synthesis was highly increased as in 12 h regenerating hepatocytes. These results indicated that in rat hepatocytes Ag-NOR proteins and stimulation of pre-rRNA synthesis are not related. The relationship between the Ag-NOR proteins and the distribution of the completely extended intranucleolar ribosomal chromatin was also studied in regenerating rat hepatocytes. At 12 h after partial hepatectomy an increased amount of completely extended ribosomal chromatin was observed, contemporaneously with an increased quantity of Ag-NOR proteins. These ribosomal chromatin changes preceded the beginning of DNA synthesis and were prevented by cycloheximide-induced inhibition of protein synthesis.
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References
Burton K (1956) A study of the condition and mechanism of the diphenylamine reaction for the colorimetric estimation of de-oxyribonucleic acid. Biochem J 62: 315–323
Cogliati R, Gautier A (1973) Mise en évidence de l'ADN et des polysaccharides a l'aide d'un nouveau réactif “de type Schiff”. CR Acad Sci Ser D 276: 3041–3044
Dabeva MD, Dudov KP, Hadjiolov HH, Stoyokova AA (1978) Quantitative analysis of rat liver nucleolar and nucleoplasmic ribosomal ribonucleic acid. Biochem J 121: 367–374
Derenzini M, Viron A, Puvion-Dutilleul F (1982a) The Feulgenlike osmium-ammine reaction as a tool to investigate chromatin structure in thin sections. J Ultrastruct Res 80: 133–147
Derenzini M, Hernandez-Verdun D, Bouteille M (1982b) Visualization in situ of DNA filaments in nuclcolar chromatin of rat hepatocytes. Exp Cell Res 141: 463–469
Derenzini M, Hernandez-Verdun D, Bouteille M (1983a) Structure of a repeating subunit organization in rat hepatocyte chromatin fixed in situ. J Cell Sci 61: 137–149
Derenzini M, Pession A, Betts-Eusebi CM, Novello F (1983b) Relationship between the extended intranucleolar chromatin in situ and ribosomal RNA synthesis. Exp Cell Res 145: 127–144
Derenzini M, Viron A, Puvion E (1984) Structural organization of rat hepatocyte chromatin as visualized in thin frozen sections selectively stain for DNA. Eur J Cell Biol 33: 148–156
Derenzini M, Farabegoli F, Pession A, Novello F (1987a) Spatial redistribution of ribosomal chromatin in the fibrillar centres of human criculating lymphocytes after stimulation of transcription. Exp Cell Res 170: 31–41
Derenzini M, Hernandez-Verdun D, Farabegoli F, Pession A, Novello F (1987b) Structure of ribosomal genes of mammalian cells in situ. Chromosoma 95: 67–70
Derenzini M, Pession A, Farabegoli F, Treré D, Badiali M, Dchan P (1989) Relationship between interphasic Nucleolar Organizer Regions and growth rate in two neuroblastoma cell lines. Am J Pathol 134: 925–932
Erard MS, Belenguer P, Caizergues-Ferrer F, Pantaloni A, Amalric F (1988) A major nucleolar protein, nucleolin, induces chromatin decondensation by binding to histone H1. Eur J Biochem 175: 515–530
Goessens G (1984) Nucleolar structure. Int Rev Cytol 84: 107–158
Hadjiolov HH, Dabeva MD, Mackendosky W (1974) The action of α-amanitin in vivo on the synthesis and maturation of mouse liver ribonucleic acid. Biochem J 138: 321–334
Hernandez-Verdun D (1983) The nucleolar organizer regions. Biol Cell 49: 191–202
Hernandez-Verdun D (1986) Structural organization of the nucleolus in mammalian cells. Methods Archiv Exp Pathol 12: 26–62
Hernandez-Verdun D, Derenzini M (1983) Non nucleosomal configuration of chromatin in nucleolar organizer regions of metaphase chromosomes in situ. Eur J Cell Biol 31: 360–365
Hernandez-Verdun D, Derenzini M, Bouteille M (1982) The morphological relationship in electron microscopy between NOR-silver proteins and intra-nucleolar chromatin. Chromosoma 85: 461–473
Hernandez-Verdun D, Derenzini M, Bouteille M (1984) Relationship between the Ag-NOR proteins and ribosomal chromatin in situ during drug-induced RNA synthesis inhibition. J Ultrastruct Res 88: 55–65
Herrera AH, Olson MOJ (1986) Association of protein C23 with rapidly labeled nucleolar RNA. Biochemistry 25: 6258–6264
Higgins GM, Anderson RA (1931) Experimental pathology of liver. 1. Restoration of liver and white rat following partial surgical removal. Arch Pathol 12: 186–202
Howell WM (1982) Selective staining of Nucleolus Organizer Regions (NORs). In: Busch H, Rothblum L (eds) The cell nucleus, vol XI. Academic Press, New York, pp 89–142
Jordan G (1987) At the heart of the nucleolus. Nature 329: 489–490
Lapeyre B, Bourbon H, Amalric F (1987) Nucleolin, the major nucleolar protein of growing eukaryotic cells: an unusual protein structure revealed by the nucleotide sequence. Proc Natl Acad Sci US 84: 1472–1476
Lischwe MA, Richards RL, Busch RK, Busch H (1981) Localization of phosphoprotein C23 to nucleolar structures and to the nucleolus organizer regions. Exp Cell Res 136: 101–109
Munro HN, Fleck A (1966) Recent developments in the measurement of nucleic acids in biological materials. Analyst 91: 78–88
Ploton D, Menager M, Jeannesson P, Himber G, Pigeon F, Adnett JJ (1986) Improvement in the staining and in the visualization of the argyrophilic proteins in the nucleolar organizer region at the optical level. Histochem J 18: 5–14
Raska I, Reimer G, Jarnik M, Kostrouch Z, Raska K (1989) Does the synthesis of ribosomal RNA take place within nucleolar fibrillar centers or dense fibrillar components? Biol Cell 76: 79–82
Reimer G, Raska I, Tan EM, Scheer U (1987) Human autoantibodies: probes for nucleolus structure and function. Virchows Arch B 54: 131–143
Scheer U, Raska I (1987) Immunocytochemical localization of RNA polymerase I in the fibrillar centers of nucleoli. Chromosomes Today 9: 284–294
Scheer U, Rose KM (1984) Localization of RNA polymerase I in interphase cells and mitotic chromosomes by light and electron microscopic immunochemistry. Proc Natl Acad Sci USA 81: 1431–1435
Thiry M, Thiry-Blaise L (1989) In situ hybridization at electron microscope level: an improved method for precise localization of ribosomal DNA and RNA. Eur J Cell Biol 50: 235–243
Verbin RS, Goldblatt PJ, Farber E (1969) The biochemical pathology of inhibition of protein synthesis in vivo. The effect of cycloheximide on hepatic parenchymal cell ultrastructure. Lab Invest 20: 179–182
Wachtler F, Hopman AHN, Wicgant J, Schwarzacher HG (1986) On the position of Nucleolus Organizer Regions (NORs) in interphase nuclei. Exp Cell Res 167: 227–240
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Pession, A., Farabegoli, F., Treré, D. et al. The Ag-NOR proteins and transcription and duplication of ribosomal genes in mammalian cell nucleoli. Chromosoma 100, 242–250 (1991). https://doi.org/10.1007/BF00344158
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DOI: https://doi.org/10.1007/BF00344158