Summary
The three-dimensional ultrastructure of epithelioid cells was studied by the quick-freezing and deepetching (QF-DE), as well as the freeze-substitution (QF-FS) methods. The granulomas were induced in rats by injecting muramyl dipeptide (MDP) into the hind footpads. At 3 weeks after the injection, the footpads were perfused with a fixative, excised, and quickly frozen to prepare the replica membranes. Some unfixed footpads were also quickly frozen and freeze-substituted. Dense networks of intermediate filaments, connected with the nuclei, mitochondria and other vesicular cell organelles, were observed throughout the cytoplasm of epithelioid cells by the QF-DE method. A few actin filaments were located in filopodia and just beneath the cell membranes. Interdigitation of the cell membranes between adjacent cells was clearly demonstrated by the QF-FS method and clathrin-coated pits were identified at the base of interdigitating filopodia. In addition, the exact moment of fusion between endosomes and lysosomes was ascertained by the same method. These results suggest that the cytoskeletal organization of epithelioid cells resembles that of epithelial cells rather than actively motile macrophages.
Similar content being viewed by others
References
Adams DO (1974) The structure of mononuclear phagocytes differentiating in vivo. Am J Pathol 76:17–48
Adams DO (1976) The granulomatous inflammatory response. Am J Pathol 84:164–192
Adams DO (1983) The biology of granuloma. In: Ioachim HL (ed) Pathology of granuloma. Raven Press, New York, pp 1–20
Baba T, Shiozawa N, Hotchi M, Ohno S (1991 a) Three-dimensional study of the cytoskeleton in macrophages and multinucleate giant cells by quick-freezing and deep-etching method. Virchows Arch [B] 61:39–47
Baba T, Shiozawa N, Hotchi M, Ohno S (1991 b) Three-dimensional identification of endosomes in macrophages by a replica scraping method. Eur J Cell Biol 56:147–150
Baum H-P, Thoenes W (1987) Freeze-fractures of epithelioid cells, multinucleated giant cells, and phagocytic macrophages. Virchows Arch[B] 53:13–22
Carr I (1980) Macrophages in connective tissues. The granuloma macrophage. In: Carr I, Daems WT (eds) The reticuloendothelial system A comprehensive treatise, vol 1. Plenum Press, New York, London, pp 671–703
Dannenberg AM Jr, Suga M, Garcia-Gonzales JE (1981) Macrophages in granulomas: Histochemical evidence suggesting local control of heterogeneous functions. In: Schmalz F, Huhn D, Schaefer HE (eds) Disorders of the monocyte macrophage system, Springer, Berlin, Heidelberg, New York, pp 109–119
Ellouz F, Adams A, Ciorbaru R, Lederer E (1974) Minimal structural requirements for adjuvant activity of bacterial peptidoglycan derivatives. Biochem Biophys Res Comm 59:1317–1325
Emori K, Tanaka A (1978) Granuloma formation by synthetic bacterial cell wall fragment: Muramyl dipeptide. Infect Immun 19:613–620
Heuser JE, Kirschner MW (1980) Filament organization revealed in platinum replicas of freeze-dried cytoskeletons. J Cell Biol 86:212–234
Ichikawa A, Ichikawa M, Hirokawa N (1980) The ultrastructure of rapid-frozen, substitution fixed parotid gland acinar cells of the mongolian gerbil. Am J Anat 157:107–110
Langer KH, Thoenes W (1981) Characterization of cells involved in the formation of granuloma. Virchows Arch [B] 36:177–194
Langer KH, Thoenes W (1984) Endocytotic activity in epithelioid and Langhans’ giant cells. Virchows Arch [B] 47:177–182
Lazarides E (1980) Intermediate filaments as mechanical integrators of cellular space. Nature 283:249–255
Leake ES, Myrvik QN (1970) Interaction of lysosome-like structures and phagosomes in normal and granulomatous alveolar macrophages. RES-J Reticuloendothel Soc 8:407–420
Marceau N, Goyette R, Deschenes J, Valet JP (1980) Morphological differences between epithelial and fibroblast cells in rat liver cultures and the role of cell surface fibronectin and cytoskeletal element organization in cell shape. Ann N Y Acad Sci 349:138–152
Naramoto A, Ohno S, Itoh N, Takami H, Nakazawa K, Shigematsu H (1990) Three-dimensional identification of actin filaments in phalloidin-treated rat livers by quick-freezing and deep-etching method. Virchows Arch [A] 417:15–20
Ohno S, Fujii Y (1991) Three-dimensional studies of cytoskeletons in cultured hepatocytes by quick-freezing and deep-etching method. Virchows Arch [A] 418:61–70
Ohno S, Takasu N (1989) Three-dimensional studies of cytoskeletal organizations in cultured thyroid cells by quick-freezing and deep-etching method. J Electron Microsc 38:352–362
Papadimitriou JM, Spector WG (1971) The origin, properties, and fate of epithelioid cells. J Pathol 105:187–203
Rapraeger A, Jalkanen M, Bernfield M (1986) Cell surface proteoglycan associates with the cytoskeleton at the basolateral cell surface of mouse mammary epithelial cells. J Cell Biol 103:2683–2696
Sugre SP, Hay ED (1981) Response of basal epithelial cell surface and cytoskeleton to solubilized extracellular matrix molecules. J Cell Biol 91:45–54
Takasu N, Ohno S, Takasu M, Yamada T (1988) Polarized thyroid cells in monolayers cultured on collagen gel: Their cytoskeleton organization, iodine uptake, and resting membrane potentials. Endocrinology 122:1021–1026
Tanaka A, Emori K (1980) Epithelioid granuloma formation by a synthetic bacterial cell wall component, muramyl dipeptide (MDP). Am J Pathol 98:733–748
Tipperman R, Kasckow J, Herndon RM (1984) The fine structure of macrophages in lysolecithin-induced demyelination: A freeze-fracture study. J Neuropathol Exp Neurol 43:522–530
Turk JL, Narayanan RB (1981) The monocyte-macrophage system in granulomatous inflammation. In: Schmalz F, Huhn D, Schaefer HE (eds) Disorders of the monocyte macrophage system, Springer, Berlin, Heidelberg, New York, pp 101–107
van der Rhee HJ, van der Burgh-de Winter CPM, Daems WT (1979) The differentiation of monocytes into macrophages, epithelioid cells, and multinucleated giant cells in subcutaneous granulomas. I. Fine structure. Cell Tiss Res 197:355–378
van Harreveld A and Crowell J (1964) Electron microscopy after rapid freezing on a metal surface and substitution fixation. Anat Rec 149:381–386
Woods A, Hook M, Kjellen L, Smith CG, Rees DA (1984) Relationship of heparan sulfate proteoglycans to the cytoskeleton and extracellular matrix of cultured fibroblasts. J Cell Biol 99:1743–1753
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Baba, T., Sakaguchi, N., Hotchi, M. et al. Three-dimensional study of epithelioid cells by a quick-freezing and deep-etching method in muramyl dipeptide-induced granulomas. Virchows Archiv B Cell Pathol 63, 63–70 (1993). https://doi.org/10.1007/BF02899245
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02899245