Summary
The aim of this study was to correlate morphological changes of nucleoli of non-proliferating monocytes to their functional activity, since nucleolar morphology is currently considered as a diagnostic marker for cell proliferation. Monocytes from healthy donors were fractionated by current counterflow centrifugation and kept in culture for 6 days. Cells were stimulated by the addition of 200 units/ml interferon γ (IFNγ). Under this stimulus the monocytes show no proliferation but a strongly augmented expression of type I Fc IgG receptor, human leucocyte antigen DR, human leucocyte antigen DP and human leucocyte antigen DQ. Morphological changes after stimulation included the appearance of multinucleated cells, typical signs of the activation of rRNA synthesis indicated by an increase in nucleolar size, and changes in nucleolar structure such as the appearance of reticulate and compact nucleoli. The number of nucleolus organiser regions (NORs) visualised by in situ hybridisation was compared with the position and number of nucleoli visualised by silverstaining in interphase cells. In comparison with control cultures, activated monocytes show a distinct increase in the number of those NORs that take part in the formation of nucleoli. Our results show that, in non-proliferating activated monocytes, the morphology of nucleoli and the increase of NOR activity are similar to those in proliferating cells. NOR activation is therefore an indicator for cellular activity, but is not necessarily correlated with proliferation.
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References
Basham TY, Merigan TC (1983) Recombinant IFNγ increases HLA-DR synthesis and expression. J Immunol 130:1492–1494
Boltz-Nitulescu G, Bazin H, Spielberg HL (1981) Specificity of Fc receptors for IgG2a, IgG1/IgG2b, and IgE on rat macrophages J Exp Med 154:374–384
Bourgeois CA, Hernandez-Verdun D, Hubert J, Bouteille M (1979) Silver staining of NORs in electron microscopy. Exp Cell Res 123:449–452
Busch H, Smetana K (1970) The nucleolus. Academic Press, New York San Francisco London
Cairns P, Suarez V, Newmann J, Crocker J (1989) Nucleolar organizer regions in transitional cell tumors of the bladder. Arch Pathol Lab Med 113:1250–1252
Derenzini M, Ploton D (1991) Interphase nucleolar organizer regions in cancer cells. Int Rev Exp Pathol 52:149–192
Derenzini M, Nardi F, Farabegoli F, Ottinetti A, Ronxaroli F, Bussolati G (1989) Distribution of silver-stained interphase nucleolar organizer regions as a parameter to distinguish neoplastic from nonneoplastic reactive cells in human effusions. Acta Cytol 33:491–498
Derenzini M, Pession A, Trere D, (1990) Quantity of nucleolar silver-stained proteins is related to proliferating activity in cancer cells. Lab Invest 63:137–140
Dougherty GJ, Dougherty ST, McBride WH (1989) Monocyte heterogeneity. In: Zembala M, Asherson GL (eds) Human monocytes. Academic Press, San Diego, pp 71–78
Dransfield I, Corcoran D, Partridge LJ, Hogg N, Burton DR (1988) Comparison of human monocytes isolated by elutriation and adherence suggests that heterogeneity may reflect a continuum of maturation/activation states. J Immunol 63:491–498
Fakan S, Puvion S (1980) Ultrastructural visulization of nucleolar and extranucleolar RNA-synthesis and distribution. Int Rew Cytol 65:255–299
Goessens G (1984) Nucleolar structure. Int Rev Cytol 87:107–158
Goodpasture C, Bloom S (1975) Visualization of nucleolar organizer regions in mammalian chromosomes using silver staining. Chromosoma 53:37–59
Hadjiolov AA (1985) The nucleolus and the ribosome biogenesis. Springer, Wien New York
Hernandez-Verdun D (1991) The nucleolus today. J Cell Sci 99:465–471
Hoover D, Meltzer M (1989) Lymphokines as monocyte activators. In: Zembala M, Asherson GL (eds) Human monocytes. Academic Press, San Diego, pp 151–160
Kay JE (1968) Early effects of phytohaemagglutinin on lymphocyte RNA-synthesis. Eur J Biochem 4:225–232
Miller DA, Dev VG, Tantravahi R, Müller OJ (1976) Suppression of human nucleolus organizer activity in mouse-human somatic hybrid cells. Exp Cell Res 101:235–243
Pinkel D, Straume T, Gray JW (1986) Cytogenetic analysis using quantitative, high-sensitivity, fluorescence hybridisation. Proc Natl Acad Sci USA 83:2934–2938
Puvion-Dutilleul F, Bachellerie JP, Puvion E (1991) Nucleolar organization of HeLa cells as studied by in situ hybridization. Chromosoma 100:395–409
Raska I, Reimer G, Jarnik M, Kostrouch Z, Raska K Jr (1989) Does the synthesis of ribosomal RNA take place within nucleolar fibrillar centers or dense fibrillar components? Biol Cell 65:79–82
Sanderson RJ, Shepperdson FT, Vatter AE, Talmage DW (1977) Isolation and enumeration of peripheral blood monocytes. J Immunol 118:1409–1414
Scheer U, Benavente R (1990) Functional and dynamic aspects of the mammalian nucleolus. Bioessays 12:14–21
Schwarzacher HG, Wachtler F (1983) Nucleolus organizer regions and nucleoli. Hum Genet 63:89–99
Schwarzacher HG, Wachtler F (1991) The functional significance of nucleolar structures. Ann Génét (Paris) 34:151–160
Schwarzacher HG, Wolf U (1974) Methods in human cytogenetics. Springer, Berlin Heidelberg New York
Schwarzacher HG, Milkesaar AV, Schnedl W (1978) The nature of Ag-staining of nucleolus organizer regions. Cytogenet Cell Genet 20:24–39
Stahl A, Wachtler F, Hartung M, Devictor M, Schöfer C, Mosgäller W, Lanversin ADE, Schwarzacher HG (1991) Nucleoli, nucleolar chromosomes and ribosomal genes in the human spermatocyte. Chromosoma 101:231–244
Wachtler F, Ellinger A, Schwarzacher HG (1980) Nucleolar changes in human phytohaemagglutinin-stimulated lymphocytes. Cell Tissue Res 213:351–360
wachtler F, Hopman HN, Wiegant J, Schwarzacher HG (1986) On the position of nucleolus organizer (NORs) in interphase nuclei. Exp Cell Res 167:227–240
Wachtler F, Hartung M, Devictor M, Wiegant J, Stahl A, Schwarzacher HG (1989) Ribosomal DNA is located and transcribed in the dense fibrillar component of human Sertoli cell nucleoli. Exp Cell Res 184:61–71
Wachtler F, Mosgöller W, Schwarzacher HG (1990a) Electron microscopic in situ hybridisation and autoradiography: localisation and transcription of rDNA in hyman lymphocyte nucleoli. Exp Cell Res 187:346–348
Wachtler F, Roubicek C, Schedle A, Mosgöller W, Bretis G, Schwarzacher HG (1990b) Nucleolus organizer regions in human lymphocytes as studied with premature chromosome condensation. Hum Genet 84:244–248
Wachtler F, Schöfer C, Mosgöller W, Weipoltshammer K, Schwarzacher HG, Guichaoua M, Hartung M, Stahl A, Berge-Lefranc JL, Gonzales I, Sylvester J (1992) Human ribosomal RNA gene repeats are localized in the dense fibrillar component of nucleoli. Light- and electron microscopic in-situ-hybridisation in human Sertoli cells. Exp Cell Res 198:135–143
Walker WS (1981) Separation of mononuclear phagocytes for enrichment and for depletion: on overview. In: Adams DO, Edelson PJ, Koren H (eds) Methods for studying mononuclear phagocytes. Academic Press, London, pp 175–178
Willheim M, Gessl A, Berger R, Schedle A, Luger T, Förster O, Boltz-Nitulescu G (1991) IL-6 augments fc IgE receptor (FcεRII/CD23) expression on human monoblastic/monocytic cell lines U 937, THP-1 and Mono-mac-6 but not on blood monocytes: regulatory effects of IL-4 and IFNγ. J Immunol 147:1837–1842
Zembala M, Asherson GL (1989) Human monocytes. Academic Press, San Diego
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Schedle, A., Willheim, M., Zeitelberger, A. et al. Nucleolar morphology and rDNA in situ hybridisation in monocytes. Cell Tissue Res 269, 473–480 (1992). https://doi.org/10.1007/BF00353902
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DOI: https://doi.org/10.1007/BF00353902