Abstract
The distributions of desmin and vimentin intermediate filaments in cultured hamster heart cells were examined by immunofluorescent microscopy and an immunogold deep-etching replica technique in combination with electron microscopy. Fluorescent studies showed the overall staining patterns of the myocytes as well as the fibroblasts. Monoclonal antibodies (Da, D3) to desmin showed punctate staining for the myocytes, while polyclonal desmin (pD) stained in a filamentous pattern. Fibroblasts stained strongly with monoclonal anti-vimentin (Va), but did not stain with the desmin probes. Deep-etched immunogold studies confirmed at the ultrastructural level that monoclonal anti-desmin antibodies stain individual intermediate filaments in an intermittent pattern. Monoclonal (D3) antibody stained the intermediate filaments heavily and continuously at the cell peripheries, while it stained intermittently in the cell body, similar to the Da monoclonal. Monoclonal anti-vimentin stained only intermediate filaments in fibroblasts. Our studies show a heterogeneity of staining within the cultured heart cells when various anti-desmin and anti-vimentin antibodies are used.
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References
Bard F, Franzini-Armstrong C (1991) Extra actin filaments at the periphery of skeletal muscle myofibrils. Tissue and Cell 23:191–197
Bennet GS, Fellini S, Toyama Y, Holtzer H (1979) Redistribution of intermediate filament subunits during skeletal myogenesis and maturation in vitro. J Cell Biol 82:577–584
Danto SI, Fischman DA (1984) Immunocytochemical analysis of intermediate filaments in embryonic heart cells with monoclonal antibodies to desmin. J Cell Biol 98:2179–2191
Dlugosz AA, Antin PB, Nachmias VT, Holtzer H (1984) The relationship between stress fiber-like structures and nascent myofibrils in cultured cardiac myocytes. J Cell Biol 99:2268–2278
Fujimoto T, Tokuyasu KT, Singer SJ (1987) Direct morphological demonstration of the coexistence of vimentin and desmin in the same intermediate filaments of vascular smooth muscle cells. J Submicrosc Cytol 19:1–9
Fuseler JW, Shay JW (1982) The association of desmin with the developing myofibrils of cultured embryonic rat heart myocytes. Dev Biol 91:448–457
Fuseler JW, Shay JW, Feit H (1981) The role of intermediate (10 nm) filaments in the development and integration of the myofibrillar contractile apparatus in the embryonic mammalian heart. In: Dowben RM, Shay JW (eds) Cell and muscle motility, vol 1. Plenum Press, New York, pp 205–259
Gard DL, Lazarides E (1980) The synthesis and distribution of desmin and vimentin during myogenesis in vitro. Cell 19:263–275
Gard DL, Bell PB, Lazarides E (1979) Co-existence of desmin and the fibroblastic intermediate filament subunit in muscle and nonmuscle cells: identification and comparative peptide analysis. Proc Natl Acad Sci USA 76:3894–3898
Hou GR, Isobe Y, Lemanski LF (1991) Immunofluorescent and immunogold replica studies of desmin distribution in cultured normal and cardiomyopathic hamster heart cells. Acta Anat 142:215–226
Ip W (1988) Modulation of desmin intermediate filament assembly by a monoclonal antibody. J Cell Biol 106:735–749
Ip W, Danto SI, Fischman DA (1983) Detection of desmin-containing intermediate filaments in cultured muscle and nonmuscle cells by immunoelectron microscopy. J Cell Biol 96:401–408
Ip W, Heuser JE, Pang Y-YS, Hartzer MK, Robson RM (1985a) Subunit structure of desmin and vimentin protofilaments and how they assemble into intermediate filaments. Ann NY Acad Sci 455:185–199
Ip W, Hartzer MK, Pang Y-YS, Robson RM (1985b) Assembly of vimentin in vitro and its implications concerning the structure of intermediate filaments. J Mol Biol 183:365–375
Ishikawa H, Bischoff R, Holtzer H (1968) Mitosis and intermediate-sized filaments in developing skeletal muscle. J Cell Biol 38:538–555
Isobe Y, Shimada Y (1986) Organization of filaments underneath the plasma membrane of developing chicken skeletal muscle cells in vitro revealed by the freeze-dry and rotary replica method. Cell Tissue Res 244:47–56
Isobe Y, Warner FD, Lemanski LF (1988) Three-dimensional immunogold localization of α-actinin within the cytoskeletal networks of cultured cardiac muscle and nonmuscle cells. Proc Natl Acad Sci USA 85:6758–6762
Isobe Y, Hou GR, Lemanski LF (1991) Deep-etching immunogold replica electron microscopy of cytoskeletal elements in cultured hamster heart cells. Anat Rec 229:415–426
Johnson DA, Gautsch JW, Sportsman JR, Elder JH (1984) Improved technique utilized nonfat dry milk for analysis of proteins and nucleic acid transferred to nitrocellulose. Gene Anal Tech 1:3–8
Kuruc N, Franke WN (1988) Transient coexpression of desmin and cytokeratins 8 and 18 in developing myocardial cells of some vertebrate species. Differentiation 38:177–193
Lazarides E (1978) The distribution of desmin (100 Å) filaments in primary cultures of embryonic chicken cardiac cells. Exp Cell Res 112:265–273
Lazarides E (1980) Intermediate filaments as mechanical integrators of cellular space. Nature 283:249–256
Lazarides E (1981) Intermediate filaments — chemical heterogeneity in differentiation. Cell 23:649–650
Lazarides E, Granger BL, Gard DL, O'Conner CM, Breckler J, Price M, Danto SI (1982) Desmin- and vimentin-containing filaments and their role in the assembly of the Z disk in muscle cells. Cold Spring Harbor Symp Quant Biol 46:351–378
Lemanski LF, Tu Z-H (1983) Immunofluorescent studies for myosin, actin, tropomyosin, and α-actinin in cultured cardiomyopathic hamster heart cells. Dev Biol 97:338–348
Mazia D, Schatten G, Sale W (1975) Adhesion of cells to surfaces coated with polylysine. Application of electron microscopy. J Cell Biol 66:198–200
Nag AC, Krehel W, Cheng M (1986) Distributions of vimentin and desmin filaments in embryonic cardiac muscle cells in culture. Cytobios 45:195–209
Osborn M, Geisler N, Shaw G, Sharp G, Weber K (1982) Intermediate filaments. Cold Spring Harbor Symp Quant Biol 46:413–429
Osinska HE, Lemanski LF (1989) Immunofluorescent localization of desmin and vimekin in developing cardiac muscle of Syrian hamster. Anat Rec 223:406–413
Parke J, Miller C, Anderton BH (1986) Higher plant myosin heavy-chain identified using a monoclonal antibody. Eur J Cell Biol 41:9–13
Quinlan RA, Franke NW (1982) Heteropolymer filaments of vimentin and desmin in vascular smooth muscle tissue and cultured baby hamster kidney cells demonstrated by chemical crosslinking. Proc Natl Acad Sci USA 79:3452–3456
Samuel JL, Jockusch B, Bertier-Savalle B, Escoubet B, Morotte F, Swynghedauw B, Rappaport L (1985) Myofibrillar organization and desmin in rat heart myocytes. Basic Res Cardiol 80:119–122
Schiller A, Taugner R (1980) Freeze-fracturing and deep-etching with the volatile cryoprotectant ethanol reveals true membrane surfaces of kidney structures. Cell Tissue Res 210:57–69
Steinert PM, Steven AC, Roop DR (1985) The molecular biology of intermediate filaments. Cell 42:411–419
Stewart M (1993) Intermediate filament structure and assembly. Curr Opin Cell Biol 5:3–11
Tokuyasu KT (1983) Visualization of longitudinally-oriented intermediate filaments in frozen sections of chicken cardiac muscle by a new staining method. J Cell Biol 97:562–565
Tokuyasu KT, Maher PA, Singer SJ (1984) Distributions of vimentin and desmin in developing chick myotubes in vivo. I. Immunofluorescence study. J Cell Biol 100:1157–1166
Tokuyasu KT, Maher PM, Singer SJ (1985) Distributions of vimentin and desmin in developing chick myotubes in vivo. II. Immunoelectron microscopic study. J Cell Biol 100:1157–1166
Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets; procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354
Tuszynski GP, Frank ED, Damsky CH, Buck CA, Warren L (1979) The detection of smooth muscle desmin-like protein in BHK-21/C-13 fibroblasts. J Biol Chem 254:6138–6143
Weber K, Geisler N (1985) Intermediate filaments: structural conservation and divergence. Ann NY Acad Sci 455:126–143
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Isobe, Y., Nakatsugawa, M., Hou, G.R. et al. Three-dimensional distributions of desmin and vimentin in cultured hamster cardiomyocytes using the immunogold deep-etching replica technique. Histochemistry 101, 155–168 (1994). https://doi.org/10.1007/BF00269541
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DOI: https://doi.org/10.1007/BF00269541