Synopsis
Fixation under ‘improper’ conditions ofin vitro cultivated cells results in an extensive diffusion of the lysosomal enzyme acid phosphatase because of the influence of a low effective osmotic pressure. In the present investigation, advantage was taken of this predictable diffusion in order to establish whether or not leakage of acid phosphatase could take place through ultrastructurally ‘intact’ lysosomal membranes.
In order to reveal small holes in the lysosomal membranes, secondary lysosomes were labelled with thorium dioxide particles, which were presumed to appear free in the cell sap if ruptures in the membranes larger than about Ioo Å were created.
The experiments revealed that following the fixation ofin vitro cultivated human glia cells under ‘improper’ conditions, mitochondria and ground cytoplasm show considerable swelling artifacts, while secondary lysosomes appear to be essentially unaffected. The lysosomes, nevertheless, apparently lost most of their content of acid phosphatase, as judged from enzyme cytochemical studies. These findings indicate that leakage of acid phosphatase from ultrastructurally ‘intact’ lysosomes is possible.
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References
Abercrombie, M. &Heaysman, J. E. M. (1953). Observations on the social behaviour of cells in tissue culture. I. Speed of movement of chick heart fibroblasts in relation to their mutual contacts.Exptl. Cell Res. 5, 111–31.
Allison, A. C. (1968). Lysosomes. In:The Biological Basis of Medicine (eds. E. E. Bittar and N. Bittar), Vol. 1, p. 209. Academic Press: London and New York.
Anderson, P. J. (1967). Purification and quantitation of glutaraldehyde and its effect on several enzyme activities in skeletal muscle.J. Histochem. Cytochem. 15, 652–61.
Arborgh, B., Dricsson, J. L. E. &Helminen, H. (1971). Inhibition of renal acid phosphatase and aryl sulfatase activity by glutaraldehyde fixation.J. Histochem. Cytochem. 19, 449–51.
Barka, T. &Anderson, P. J. (1962). Histochemical methods for acid phosphatase using hexazonium pararosanilin as a coupler.J. Histochem. Cytochem. 10, 741–53.
Berthet, J., Berthet, L., Appelmans, F. &de Duve, C. (1951). Tissue fractionation studies. 2. The nature of the linkage between acid phosphatase and mitochondria in rat-liver tissue.Biochem. J. 50, 182–9.
Biberfeld, P. (1968). A method for the study of monolayer cultures with preserved cell orientation and interrelationship.J. Ultrastr. Res. 25, 158–9.
Brunk, U., Dricsson, J. L. E., Pontén, J., &Westermark, B. (1971). Specialization of cell surfaces in contact-inhibited human glia-like cellsin vitro.Exptl. Cell Res. 67, 497–15.
Brunk, U. &Dricsson, J. I. E. (1972). Demonstration of acid phosphatase inin vitro cultured cells. Significance of fixation, tonicity and permeability factors.Histochem. J. 4, 349–63.
de Duve, C. (1959). Lysosomes, a new group of cytoplasmic particles. In:Subcellular Particles, p. 128. (ed. T. Hayashi) New York: The Ronald Press Co.
de Duve, C. &Wattiaux, R. (1966). Functions of lysosomes.Ann. Rev. Physiol. 28, 435–92.
Dricsson, J. L. E. &Brunk, U. (1972). Alterations in lysosomal membranes as related to diseases processes. In:Membrane Pathology (eds. B. F. Trump and A. Arstila). New York: Academic Press. In Press.
Hayflick, L. (1965). The limitedin vitro lifetime of human diploid cell strains.Exptl. Cell Res. 37, 614–36.
Mollenhauer, H. H. (1964). Plastic embedding mixtures for use in electron microscopy.Stain Technol. 39, 111–14.
Pontén, J., Westermark, B. &Hugosson, R. (1969). Regulation of proliferation and movement of human glia-like cells in culture.Exptl. Cell Res. 58, 393–400.
Reynolds, E. S. (1963). The use of lead citrate at high pH as an electron-opaque stain in electron microscopy.J. Cell Biol. 17, 208–12.
Smith, R. E. &Farquhar, M. G. (1966). Lysosome function in the regulation of the secretory process in the cells of the anterior pituitary gland.J. Cell Biol. 31, 319–47.
Weissmann, G. (1969). The effects of steroids and drugs on lysosomes. In:Lysosomes in Biology and Pathology. (eds. J. T. Dingle and H. S. Fell), Vol. 1, p. 276. Amsterdam, London: North-Holland.
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Brunk, U.T., Dricsson, J.L.E. Cytochemical evidence for the leakage of acid phosphatase through ultrastructurally intact lysosomal membranes. Histochem J 4, 479–491 (1972). https://doi.org/10.1007/BF01011128
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DOI: https://doi.org/10.1007/BF01011128