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. 1983 Feb 1;96(2):401–408. doi: 10.1083/jcb.96.2.401

Detection of desmin-containing intermediate filaments in cultured muscle and nonmuscle cells by immunoelectron microscopy

PMCID: PMC2112300  PMID: 6339515

Abstract

Antibodies raised against chicken gizzard smooth muscle desmin were shown to be specific by immunofluorescence cytochemistry and immunoautoradiography after two-dimensional polyacrylamide gel electrophoresis. Embryonic chick heart cell cultures (permeabilized with Triton X-100) and enucleated adult chicken erythrocyte ghosts (Granger, B. L., E. A. Rapasky, and E. Lazarides, 1982, J. Cell Biol. 92:299-312) were then used for immunoelectronmicroscopic localization of desmin. As expected, all intermediate filaments (IF) of the cardiac myocytes were labeled heavily and uniformly with the desmin antibodies. No periodicity or helicity was detectable along the labeled IF. Of interest was the intermittent but clear labeling of the IF of the nonmuscle, fibroblastic cells in the identical cultures. These antibodies did not bind vimentin from embryonic chick heart homogenates; furthermore, they did not label IF of avian erythrocytes known to contain vimentin but not desmin. We conclude that IF of cardiac fibroblastic cells contain low, but significant, concentrations of desmin and that this protein probably forms a copolymer with vimentin in these cells.

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Selected References

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  1. Campbell G. R., Chamley-Campbell J., Gröschel-Stewart U., Small J. V., Anderson P. Antibody staining of 10-nm (100-A) filaments in cultured smooth, cardiac and skeletal muscle cells. J Cell Sci. 1979 Jun;37:303–322. doi: 10.1242/jcs.37.1.303. [DOI] [PubMed] [Google Scholar]
  2. Clark W. A., Jr Selective control of fibroblast proliferation and its effect on cardiac muscle differentiation in vitro. Dev Biol. 1976 Sep;52(2):263–282. doi: 10.1016/0012-1606(76)90245-1. [DOI] [PubMed] [Google Scholar]
  3. Dellagi K., Brouet J. C., Perreau J., Paulin D. Human monoclonal IgM with autoantibody activity against intermediate filaments. Proc Natl Acad Sci U S A. 1982 Jan;79(2):446–450. doi: 10.1073/pnas.79.2.446. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fellini S. A., Bennett G. S., Holtzer H. Selective binding of antibody against gizzard 10-nm filaments to different cell types in myogenic cultures. Am J Anat. 1978 Nov;153(3):451–457. doi: 10.1002/aja.1001530308. [DOI] [PubMed] [Google Scholar]
  5. Frank E. D., Warren L. Aortic smooth muscle cells contain vimentin instead of desmin. Proc Natl Acad Sci U S A. 1981 May;78(5):3020–3024. doi: 10.1073/pnas.78.5.3020. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Franke W. W., Schmid E., Weber K., Osborn M. HeLa cells contain intermediate-sized filaments of the prekeratin type. Exp Cell Res. 1979 Jan;118(1):95–109. doi: 10.1016/0014-4827(79)90587-1. [DOI] [PubMed] [Google Scholar]
  7. Friedlander M., Fischman D. A. Immunological studies of the embryonic muscle cell surface. Antiserum to the prefusion myoblast. J Cell Biol. 1979 Apr;81(1):193–214. doi: 10.1083/jcb.81.1.193. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fuseler J. W., Shay J. W. The association of desmin with the developing myofibrils of cultured embryonic rat heart myocytes. Dev Biol. 1982 Jun;91(2):448–457. doi: 10.1016/0012-1606(82)90051-3. [DOI] [PubMed] [Google Scholar]
  9. Gard D. L., Bell P. B., Lazarides E. Coexistence of desmin and the fibroblastic intermediate filament subunit in muscle and nonmuscle cells: identification and comparative peptide analysis. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3894–3898. doi: 10.1073/pnas.76.8.3894. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gard D. L., Lazarides E. The synthesis and distribution of desmin and vimentin during myogenesis in vitro. Cell. 1980 Jan;19(1):263–275. doi: 10.1016/0092-8674(80)90408-0. [DOI] [PubMed] [Google Scholar]
  11. Geisler N., Plessmann U., Weber K. Related amino acid sequences in neurofilaments and non-neural intermediate filaments. Nature. 1982 Apr 1;296(5856):448–450. doi: 10.1038/296448a0. [DOI] [PubMed] [Google Scholar]
  12. Geisler N., Weber K. Comparison of the proteins of two immunologically distinct intermediate-sized filaments by amino acid sequence analysis: desmin and vimentin. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4120–4123. doi: 10.1073/pnas.78.7.4120. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Granger B. L., Lazarides E. Desmin and vimentin coexist at the periphery of the myofibril Z disc. Cell. 1979 Dec;18(4):1053–1063. doi: 10.1016/0092-8674(79)90218-6. [DOI] [PubMed] [Google Scholar]
  14. Granger B. L., Lazarides E. Synemin: a new high molecular weight protein associated with desmin and vimentin filaments in muscle. Cell. 1980 Dec;22(3):727–738. doi: 10.1016/0092-8674(80)90549-8. [DOI] [PubMed] [Google Scholar]
  15. Granger B. L., Repasky E. A., Lazarides E. Synemin and vimentin are components of intermediate filaments in avian erythrocytes. J Cell Biol. 1982 Feb;92(2):299–312. doi: 10.1083/jcb.92.2.299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hubbard B. D., Lazarides E. Copurification of actin and desmin from chicken smooth muscle and their copolymerization in vitro to intermediate filaments. J Cell Biol. 1979 Jan;80(1):166–182. doi: 10.1083/jcb.80.1.166. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Huiatt T. W., Robson R. M., Arakawa N., Stromer M. H. Desmin from avian smooth muscle. Purification and partial characterization. J Biol Chem. 1980 Jul 25;255(14):6981–6989. [PubMed] [Google Scholar]
  18. Hunter W. M., Greenwood F. C. A radio-immunoelectrophoretic assay for human growth hormone. Biochem J. 1964 Apr;91(1):43–56. doi: 10.1042/bj0910043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lazarides E., Hubbard B. D. Immunological characterization of the subunit of the 100 A filaments from muscle cells. Proc Natl Acad Sci U S A. 1976 Dec;73(12):4344–4348. doi: 10.1073/pnas.73.12.4344. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lazarides E. Intermediate filaments as mechanical integrators of cellular space. Nature. 1980 Jan 17;283(5744):249–256. doi: 10.1038/283249a0. [DOI] [PubMed] [Google Scholar]
  21. Lazarides E. Intermediate filaments: a chemically heterogeneous, developmentally regulated class of proteins. Annu Rev Biochem. 1982;51:219–250. doi: 10.1146/annurev.bi.51.070182.001251. [DOI] [PubMed] [Google Scholar]
  22. Lazarides E. The distribution of desmin (100 A) filaments in primary cultures of embryonic chick cardiac cells. Exp Cell Res. 1978 Mar 15;112(2):265–273. doi: 10.1016/0014-4827(78)90209-4. [DOI] [PubMed] [Google Scholar]
  23. Moss M., Asch B., Schwartz R. Differentiation of actin-containing filaments during chick skeletal myogenesis. Exp Cell Res. 1979 Jun;121(1):167–178. doi: 10.1016/0014-4827(79)90457-9. [DOI] [PubMed] [Google Scholar]
  24. Osborn M., Franke W., Weber K. Direct demonstration of the presence of two immunologically distinct intermediate-sized filament systems in the same cell by double immunofluorescence microscopy. Vimentin and cytokeratin fibers in cultured epithelial cells. Exp Cell Res. 1980 Jan;125(1):37–46. doi: 10.1016/0014-4827(80)90186-x. [DOI] [PubMed] [Google Scholar]
  25. Osborn M., Ludwig-Festl M., Weber K., Bignami A., Dahl D., Bayreuther K. Expression of glial and vimentin type intermediate filaments in cultures derived from human glial material. Differentiation. 1981;19(3):161–167. doi: 10.1111/j.1432-0436.1981.tb01143.x. [DOI] [PubMed] [Google Scholar]
  26. Pruss R. M., Mirsky R., Raff M. C., Thorpe R., Dowding A. J., Anderton B. H. All classes of intermediate filaments share a common antigenic determinant defined by a monoclonal antibody. Cell. 1981 Dec;27(3 Pt 2):419–428. doi: 10.1016/0092-8674(81)90383-4. [DOI] [PubMed] [Google Scholar]
  27. Quinlan R. A., Franke W. W. 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 U S A. 1982 Jun;79(11):3452–3456. doi: 10.1073/pnas.79.11.3452. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Renner W., Franke W. W., Schmid E., Geisler N., Weber K., Mandelkow E. Reconstitution of intermediate-sized filaments from denatured monomeric vimentin. J Mol Biol. 1981 Jun 25;149(2):285–306. doi: 10.1016/0022-2836(81)90303-x. [DOI] [PubMed] [Google Scholar]
  29. Schmid E., Osborn M., Rungger-Brändle E., Gabbiani G., Weber K., Franke W. W. Distribution of vimentin and desmin filaments in smooth muscle tissue of mammalian and avian aorta. Exp Cell Res. 1982 Feb;137(2):329–340. doi: 10.1016/0014-4827(82)90034-9. [DOI] [PubMed] [Google Scholar]
  30. Schnitzer J., Franke W. W., Schachner M. Immunocytochemical demonstration of vimentin in astrocytes and ependymal cells of developing and adult mouse nervous system. J Cell Biol. 1981 Aug;90(2):435–447. doi: 10.1083/jcb.90.2.435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Small J. V., Sobieszek A. Studies on the function and composition of the 10-NM(100-A) filaments of vertebrate smooth muscle. J Cell Sci. 1977 Feb;23:243–268. doi: 10.1242/jcs.23.1.243. [DOI] [PubMed] [Google Scholar]
  32. Steinert P. M., Idler W. W., Cabral F., Gottesman M. M., Goldman R. D. In vitro assembly of homopolymer and copolymer filaments from intermediate filament subunits of muscle and fibroblastic cells. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3692–3696. doi: 10.1073/pnas.78.6.3692. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Tuszynski G. P., Frank E. D., Damsky C. H., Buck C. A., Warren L. The detection of smooth muscle desmin-like protein in BHK21/C13 fibroblasts. J Biol Chem. 1979 Jul 10;254(13):6138–6143. [PubMed] [Google Scholar]
  35. Wang E., Cross R. K., Choppin P. W. Involvement of microtubules and 10-nm filaments in the movement and positioning of nuclei in syncytia. J Cell Biol. 1979 Nov;83(2 Pt 1):320–337. doi: 10.1083/jcb.83.2.320. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Webster R. E., Henderson D., Osborn M., Weber K. Three-dimensional electron microscopical visualization of the cytoskeleton of animal cells: immunoferritin identification of actin- and tubulin-containing structures. Proc Natl Acad Sci U S A. 1978 Nov;75(11):5511–5515. doi: 10.1073/pnas.75.11.5511. [DOI] [PMC free article] [PubMed] [Google Scholar]

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