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Electron microscopic and enzyme cytochemical studies on the guinea pig metaphysis with special reference to the lysosomal system of different cell types

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Summary

A transmission electron microscopic study of demineralized, methaphyseal bone of the young guinea pig is presented. Special attention is paid to the lysosomal system of the different cell types. Visualization of acid phosphatase and aryl sulfatase activity was used to identify tissue components as belonging thereto. The distribution of alkaline phosphatase activity, a plasma membrane marker, was also examined.

Osteoblasts were distinguished by a marked development of the granular endoplasmic reticulum and the Golgi complex. Perivascular cells type A, morphologically resembled the osteoblasts, and are believed to represent an early stage in the specialization of the latter. A few lysosomes were normally found in the osteoblasts; they were less common in the type A cells. In contrast to their regular occurrence in guinea pig epiphyseal cartilage, dense bodies of lysosomal nature (“type I vesicles”) were only rarely seen in the bone matrix. Structures analogous to the type II vesicles in cartilage were, however, normally present. Their membrane showed activity of alkaline phosphatase. Possible functions of lysosomes and matrix vesicles in osteogenesis are discussed.

Perivascular cells type B and chondroclasts both contained a prominent Golgi complex and large numbers of free ribosomes, mitochondria and lysosomes. In the type B cells, inclusion material of varying appearance often occurred in the lysosomes and in endocytic vesicles. The chondroclasts sometimes presented a ruffled border, with associated vacuoles and lysosomes in the subjacent cytoplasm. It is suggested that both cell types participate in the resorption of the epiphyseal cartilage. Chondroclasts presumably arise by fusion of type B cells and/or monocytic precursors from the peripheral blood.

The skilled technical assistance of Mrs. Eva Lundberg and the secretarial assistance of Mrs. Inger Åhrén are gratefully acknowledged.

The authors are indebted to Dr. Stanislaw Moskalewski for constructive criticism of the manuscript.

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References

  • Anderson, H. C., Reynolds, J. J.: Pyrophosphate stimulation of calcium uptake into cultured embryonic bones. Fine structure of matrix vesicles and their role in calcification. Develop. Biol. 34, 211–227 (1973)

    Google Scholar 

  • Barrett, A. J.: Lysosomal enzymes. In: Lysosomes, a laboratory handbook (J. T. Dingle, ed.), p. 46–135. Amsterdam-London: North-Holland Publ. 1972

    Google Scholar 

  • Baylink, D., Wergedal, J., Thompson, E.: Loss of proteinpolysaccharides at sites where bone mineralization is initiated. J. Histochem. Cytochem. 20, 279–292 (1972)

    Google Scholar 

  • Beck, F., Lloyd, J. B., Squier, C. A.: Histochemistry. In: Lysosomes, a laboratory handbook (J. T. Dingle, ed.), p. 200–239. Amsterdam-London: North-Holland Publ. 1972

    Google Scholar 

  • Bohman, S.-O., Maunsbach, A. B.: Effects on tissue fine structure of variations in colloid osmotic pressure of glutaraldehyde fixatives. J. Ultrastruct. Res. 30, 195–208 (1970)

    Google Scholar 

  • Cameron, D. A.: The ultrastructure of bone. In: The biochemistry and physiology of bone, vol. 1: Structure (G. H. Bourne, ed.), p. 191–236. New York and London: Academic Press 1972

    Google Scholar 

  • Cliff, W. J.: Observations on healing tissue: A combined light and electron microscopic investigation. Phil. Trans. B 246, 305–325 (1963)

    Google Scholar 

  • Daems, W. Th., Wisse, E., Brederoo, P.: Electron microscopy of the vacuolar apparatus. In: Lysosomes, a laboratory handbook (J. T. Dingle, ed.), p. 150–199. Amsterdam-London: North-Holland Publ. 1972

    Google Scholar 

  • Dingle, J. T.: The extracellular secretion of lysosomal enzymes. In: Lysosomes in biology and pathology, vol. 2 (J. T. Dingle, H. B. Fell, eds.), p. 421–436. Amsterdam-London: NorthHolland Publ. 1969

    Google Scholar 

  • Dingle, J. T., Barrett, A. J., Weston, P. D.: Cathepsin D. Characteristics of immunoinhibition and the confirmation of a role in cartilage breakdown. Biochem. J. 123, 1–13 (1971)

    Google Scholar 

  • Doty, S. B., Schofield, B. H., Robinson, R. A.: The electron microscopic identification of acid phosphatase and adenosinetriphosphatase in bone cells following parathyroid extract or thyrocalcitonin administration. Excerpta ed. (Amst.) 14, 169–181 (1968)

    Google Scholar 

  • Ericsson, J. L. E.: Mechanism of cellular autophagy. In: Lysosomes in biology and pathology, vol.2 (J. T. Dingle, H. B. Pell, eds.), p. 345–394. Amsterdam-London: North-Holland Publ. 1969

    Google Scholar 

  • Evanson, J. M.: Mammalian collagenases and their role in connective tissue breakdown. In: Tissue proteinases (A. J. Barrett, J. T. Dingle, eds.), p. 327–342. Amsterdam-London: North-Holland Publ. 1971

    Google Scholar 

  • Farquhar, M. G., Palade, G. E.: Junctional complexes in various epithelia. J. Cell Biol. 17, 375–412 (1963)

    Google Scholar 

  • Furth, R. van, Cohn, Z. A., Hirsch, J. G., Humphrey, J. H., Spector, W. G., Langevoort, H. L.: The mononuclear phagocyte system: a new classification of macrophages, monoLucht, U., Maunsbach, A. B.: Effects of parathyroid hormone on osteoclasts in vivo. An ultrastructural and histochemical study. Z. Zellforsch. 141, 529–544 (1973)

    Google Scholar 

  • Göthlin, G.: Electron microscopic observations on fracture repair in the rat. Acta path. microbiol. scand. A. 81, 507–522 (1973)

    Google Scholar 

  • Göthlin, G., Ericsson, J. L. E.: Observations on the mode of uptake of thorium dioxide particles by osteoclasts in fracture callus. Calcif. Tiss. Bes. 10, 216–222 (1972)

    Google Scholar 

  • Göthlin, G., Ericsson, J. L. E.: On the histogenesis of the cells in fracture callus. Electron microscopic autoradiographic observations in parabiotic rats and studies on labeled monocytes. Virchows Arch. Abt. B Zellpath. 12, 318–329 (1973a)

    Google Scholar 

  • Göthlin, G., Ericsson, J. L. E.: Fine structural localization of acid phosphomonoesterase in the osteoblasts and osteocytes of fracture callus. Histochemie 35, 81–91 (1973b)

    Google Scholar 

  • Göthlin, G., Ericsson, J. L. E.: Electron microscopic studies on the uptake and storage of thorium dioxide molecules in different cell types of fracture callus. Acta path. microbiol. scand. A 81, 523–542 (1973c)

    Google Scholar 

  • Goldfischer, S.: The cytochemical demonstration of lysosomal aryl sulfatase activity by light and electron microscopy. J. Histochem. CytoChem. 13, 520–523 (1965)

    Google Scholar 

  • Hancox, N. M.: The osteoclast. In: The biochemistry and physiology of bone, vol. 1: Structure (G. H. Bourne, ed.), p. 45–67. New York and London: Academic Press 1972

    Google Scholar 

  • Herring, G. M.: The organic matrix of bone. In: The biochemistry and physiology of bone, vol. 1: Structure (G. H. Bourne, ed.), p. 127–189. New York and London: Academic Press 1972

    Google Scholar 

  • Leduc, E. H., Bernhard, W., Holt, S. J., Tranzer, J. P.: Ultrathin frozen sections. II. Demonstration of enzymic activity. J. Cell Biol. 34, 773–786 (1967)

    Google Scholar 

  • Lucht, U.: Absorption of peroxidase by osteoclasts as studied by electron microscope histochemistry. Histochemie 29, 274–286 (1972a)

    Google Scholar 

  • Lucht, U.: Osteoclasts and their relationship to bone as studied by electron microscopy. Z. Zellforsch. 135, 211–228 (1972b)

    Google Scholar 

  • Lucht, U.: Cytoplasmic vacuoles and bodies of the osteoclast. An electron microscope study. Z. Zellforsch. 135, 229–244 (1972c)

    Google Scholar 

  • Lucht, U., Maunsbach, A. B.: Effects of parathyroid hormone on osteoclasts in vivo. An ultrastructural and histochemical study. Z. Zellforsch. 141, 529–544 (1973)

    Google Scholar 

  • Luft, J. H.: Improvements in epoxy resin embedding methods. J. biophys. biochem. Cytol. 9, 409–414 (1961)

    Google Scholar 

  • Mayahara, H., Hirano, H., Saito, T., Ogawa, K.: The new lead citrate method for the ultracytochemical demonstration of activity of nonspecific alkaline phosphatase (orthophosphoric monoester phosphohydrolase). Histochemie 11, 88–96 (1967)

    Google Scholar 

  • Miller, E. J.: A review of biochemical studies on the genetically distinct collagens of the skeletal system. Clin. Orthop. Related Res. 92, 260–280 (1973)

    Google Scholar 

  • Morrison, R. I. G., Barrett, A. J., Dingle, J. T., Prior, D.: Cathepsins BI and D. Action on human cartilage proteoglycans. Biochim. biophys. Acta (Amst.) 302, 411–419 (1973)

    Google Scholar 

  • Mueller, W. J., Brubaker, R. L., Gay, C. V., Boelkins, J. N.: Mechanisms of bone resorption in laying hens. Fed. Proc. 32, 1951–1954 (1973)

    Google Scholar 

  • Nichols, G.: In vitro studies of bone resorptive mechanisms. In: Mechanisms of hard tissue destruction (R. F. Sognnaes, ed.), p. 557–575. Washington, D. C.: American association for the advancement of science 1963.

  • Nordwig, A.: Collagenolytic enzymes. Advanc. Enzymol. 34, 155–205 (1971)

    Google Scholar 

  • Pugliarello, M. C., Vittur, F., Bernard, B. de, Bonucci, E., Ascenzi, A.: Chemical modifications in osteones during calcification. Calcif. Tiss. Bes. 5, 108–114 (1970)

    Google Scholar 

  • Revel, J. P.: A stain for the ultrastructural localization of acid mucopolysaccharides. J. Microsc. (Paris) 3, 535–544 (1964)

    Google Scholar 

  • Reynolds, E. S.: The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J. Cell Biol. 17, 208–212 (1963)

    Google Scholar 

  • Schenk, R. K., Spiro, D., Wiener, J.: Cartilage resorption in the tibial epiphyseal plate of growing rats. J. Cell Biol. 34, 275–291 (1967)

    Google Scholar 

  • Schenk, R. K., Wiener, J., Spiro, D.: Fine structural aspects of vascular invasion of the tibial epiphyseal plate of growing rats. Acta anat. (Basel) 69, 1–17 (1968)

    Google Scholar 

  • Scherft, J. P.: The ultrastructure of the organic matrix of calcified cartilage and bone in embryonic mouse radii. J. Ultrastruct. Res. 23, 333–343 (1968)

    Google Scholar 

  • Scherft, J. P.: The lamina limitans of the organic matrix of calcified cartilage and bone. J. Ultrastruct. Res. 38, 318–331 (1972)

    Google Scholar 

  • Schoefl, G. I.: Studies on inflammation. III. Growing capillaries: Their structure and permeability. Virchows Arch. path. Anat. 337, 97–141 (1963)

    Google Scholar 

  • Scott, B. L.: Thymidine-3H electron microscope radioautography of osteogenic cells in the fetal rat. J. Cell Biol. 35, 115–126 (1967)

    Google Scholar 

  • Scott, B. L., Glimcher, M. J.: Distribution of glycogen in osteoblasts of the fetal rat. J. Ultrastruct. Res. 36, 565–586 (1971)

    Google Scholar 

  • Sjöstrand, F. S.: Electron microscopy of cells and tissues, vol.1: Instrumentation and techniques, p. 145. New York-London: Academic Press 1967

    Google Scholar 

  • Spurr, A. R.: A low-viscosity epoxy resin embedding medium for electron microscopy. J. Ultrastruct. Res. 26, 31–43 (1969)

    Google Scholar 

  • Thyberg, J.: Ultrastructural localization of aryl sulfatase activity in the epiphyseal plate. J. Ultrastruct. Res. 38, 332–342 (1972)

    Google Scholar 

  • Thyberg, J: Electron microscopic studies on the initial phases of calcification in guinea pig epiphyseal cartilage. J. Ultrastruct. Res. 46, 206–218 (1974)

    Google Scholar 

  • Thyberg, J.: Electron microscopic studies on the uptake of exogenous marker particles by different cell types in the guinea pig metaphysis. Cell Tissue Res. 156, 301–315 (1975)

    Google Scholar 

  • Thyberg, J., Friberg, U.: Ultrastructure and acid phosphatase activity of matrix vesicles and cytoplasmic dense bodies in the epiphyseal plate. J. Ultrastruct. Res. 33, 554–573 (1970)

    Google Scholar 

  • Thyberg, J., Friberg, U.: Electron microscopic enzyme histochemical studies on the cellular genesis of matrix vesicles in the epiphyseal plate. J. Ultrastruct. Res. 41, 43–59 (1972)

    Google Scholar 

  • Thyberg, J., Lohmander, S., Friberg, U.: Electron microscopic demonstration of proteoglycans in guinea pig epiphyseal cartilage. J. Ultrastruct. Res. 45, 407–427 (1973)

    Google Scholar 

  • Vaes, G.: On the mechanisms of bone resorption. The action of parathyroid hormone on the excretion and synthesis of lysosomal enzymes and on the extracellular release of acid by bone cells. J. Cell Biol. 39, 676–697 (1968)

    Google Scholar 

  • Vaes, G.: Lysosomes and the cellular physiology of bone resorption. In: Lysosomes in biology and pathology, vol. 1 (J. T. Dingle, H. B. Fell, eds.), p. 217–253. Amsterdam-London: North-Holland Publ. 1969

    Google Scholar 

  • Watson, M. L.: Staining of tissue sections for electron microscopy with heavy metals. J. biophys. biochem. Cytol. 4, 475–478 (1958)

    Google Scholar 

  • Woessner, J. F., Jr.: Purification of cathepsin D from cartilage and uterus and its action on the protein-polysaccharide complex of cartilage. J. biol. Chem. 248, 1634–1642 (1973a)

    Google Scholar 

  • Woessner, J. F., Jr.: Mammalian collagenases. Clin. Orthop. Related Res. 96, 310–326 (1973b)

    Google Scholar 

  • Yoshiki, S., Umeda, T., Kurahashi, Y.: An effective reactivation of alkaline phosphatase in hard tissues completely decalcified for light and electron microscopy. Histochemie 29, 296–304 (1972)

    Google Scholar 

  • Young, R. W.: Cell proliferation and specialization during endochondral osteogenesis in young rats. J. Cell Biol. 14, 357–370 (1962)

    Google Scholar 

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Authors and Affiliations

  1. Department of Histology, Karolinska Institutet, Stockholm, Sweden

    Johan Thyberg, Stefan Nilsson & Ulf Friberg

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  1. Johan Thyberg
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  2. Stefan Nilsson
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  3. Ulf Friberg
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Financial support was obtained from the Swedish Medical Research Council (proj. no. 12X-3355), the Swedish Cancer Society (proj. no. 100-K71-05XK), the King Gustaf V 80th Birthday Fund, the Harald and Greta Jeansson Foundations, the C. B. Nathhorst Foundation, and from the funds of Karolinska Institutet.

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Thyberg, J., Nilsson, S. & Friberg, U. Electron microscopic and enzyme cytochemical studies on the guinea pig metaphysis with special reference to the lysosomal system of different cell types. Cell Tissue Res. 156, 273–299 (1975). https://doi.org/10.1007/BF00225359

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