Abstract
The brilliantly fluorescent cytoplasmic particles that accumulate in HeLa cells treated with acridine orange, previously referred to as acridine orange particles, are shown to represent acid phosphatase positive multivesicular bodies (MVB). Dynamic changes in the ultrastructure of these organelles may be induced by varying the concentration of extracellular dye and the length of exposure to the dye. Low concentrations of dye for long intervals of time lead to marked hypertrophy of the MVB and accumulation of myelin figures within them, the acid phosphatase activity being retained. High concentrations of dye for short time intervals lead initially to a diffuse distribution of dye through out the cytoplasm (cytoplasmic reddening) as viewed in the fluorescence microscope. When cells are stained in this way and incubated in a dye-free medium, the diffusely distributed dye is segregated into MVB within 1 hour. Ultrastructurally, these MVB show dilatation but no myelin figures. The process of dye segregation is energy dependent and will not occur in starved cells. This energy dependence and the occurrence of segregation via dilatation of the MVB rather than ultrastructural transformation, i.e. formation of new binding sites, suggests that the process involves an active transport mechanism. Of the various energy sources supplied to starved cells, only glucose, mannose, and pyruvate are fully effective in supporting dye segregation. Blockage of the tricarboxylic acid cycle with malonate inhibits the effects of pyruvate but not of glucose, demonstrating the efficacy of both the tricarboxylic acid and glycolytic cycles in supplying energy for the process.
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Selected References
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- GOMORI G. Histochemical methods for acid phosphatase. J Histochem Cytochem. 1956 Sep;4(5):453–461. doi: 10.1177/4.5.453. [DOI] [PubMed] [Google Scholar]
- NOVIKOFF A. B., BEAUFAY H., DE DUVE C. Electron microscopy of lysosomerich fractions from rat liver. J Biophys Biochem Cytol. 1956 Jul 25;2(4 Suppl):179–184. [PMC free article] [PubMed] [Google Scholar]
- OGAWA K., MIZUNO N., OKAMOTO M. Lysosomes in cultured cells. J Histochem Cytochem. 1961 Mar;9:202–202. doi: 10.1177/9.2.202. [DOI] [PubMed] [Google Scholar]
- REBHUN L. I. Aster-associated particles in the cleavage of marine invertebrate eggs. Ann N Y Acad Sci. 1960 Oct 7;90:357–380. doi: 10.1111/j.1749-6632.1960.tb23257.x. [DOI] [PubMed] [Google Scholar]
- ROBBINS E., MARCUS P. I. DYNAMICS OF ACRIDINE ORANGE-CELL INTERACTION. I. INTERRELATIONSHIPS OF ACRIDINE ORANGE PARTICLES AND CYTOPLASMIC REDDENING. J Cell Biol. 1963 Aug;18:237–250. doi: 10.1083/jcb.18.2.237. [DOI] [PMC free article] [PubMed] [Google Scholar]
- ROBBINS E. The rate of proflavin passage into single living cells with application to permeability studies. J Gen Physiol. 1960 Mar;43:853–866. doi: 10.1085/jgp.43.4.853. [DOI] [PMC free article] [PubMed] [Google Scholar]
- SCHMIDT W. [Photo- and electron microscopic studies on the intracellular processing of vital stains]. Z Zellforsch Mikrosk Anat. 1962;58:573–637. [PubMed] [Google Scholar]
- STOECKENIUS W. An electron microscope study of myelin figures. J Biophys Biochem Cytol. 1959 May 25;5(3):491–500. doi: 10.1083/jcb.5.3.491. [DOI] [PMC free article] [PubMed] [Google Scholar]
- WITTEKIND D. Die Vitalfărbung des Măuseasciteskarzinoms mit Acridinorange. Z Zellforsch Mikrosk Anat. 1958;49(1):58–104. [PubMed] [Google Scholar]
- WITTEKIND D., VOLCKER A. Untersuchungen an Ergüssen seröser Höhlen mit dem kombinierten Phasenkontrast-Fluorescenz-Mikroskop. Z Klin Med. 1958;155(5):403–417. [PubMed] [Google Scholar]
- WOLF M. K., ARONSON S. B. Growth fluorescence and metachromasy of cells cultured in the presence of acridine orange. J Histochem Cytochem. 1961 Jan;9:22–29. doi: 10.1177/9.1.22. [DOI] [PubMed] [Google Scholar]