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. 1968 Aug 1;38(2):392–402. doi: 10.1083/jcb.38.2.392

AUTOPHAGIC VACUOLES PRODUCED IN VITRO

II. Studies on the Mechanism of Formation of Autophagic Vacuoles Produced by Chloroquine

Martha E Fedorko 1, James G Hirsch 1, Zanvil A Cohn 1
PMCID: PMC2107481  PMID: 4874492

Abstract

Continuous phase-contrast observations have been made on macrophages following exposure to chloroquine. The initial abnormality is the appearance in the Golgi region of small vacuoles with an intermediate density between that of pinosomes and granules. Over the course of 1–2 hr these vacuoles grow larger and accumulate amorphous material or lipid. Pinosomes or granules frequently fuse with the toxic vacuoles. Chloroquine derivatives can be seen by fluorescence microscopy; the drug is rapidly taken up by macrophages and localized in small foci in the Golgi region. Chloroquine continues to produce vacuoles when pinocytosis is suppressed. Electron microscopic studies of chloroquine effects on macrophages preincubated with colloidal gold to label predominately pinosomes or granules suggest that toxic vacuoles can arise from unlabeled organelles. Later vacuoles regularly acquire gold label, apparently by fusion, from both granules and pinosomes. L cells also develop autophagic vacuoles after exposure to chloroquine. Smooth endoplasmic reticulum apparently is involved early in the autophagic process in these cells. Information now available suggests an initial action of chloroquine on Golgi or smooth endoplasmic reticulum vesicles, and on granules, with alterations in their membranes leading to fusion with one another and with pinosomes.

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Selected References

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  1. ALLISON A. C., YOUNG M. R. UPTAKE OF DYES AND DRUGS BY LIVING CELLS IN CULTURE. Life Sci. 1964 Dec;3:1407–1414. doi: 10.1016/0024-3205(64)90082-7. [DOI] [PubMed] [Google Scholar]
  2. Allison J. L., O'Brien R. L., Hahn F. E. DNA: reaction with chloroquine. Science. 1965 Sep 3;149(3688):1111–1113. doi: 10.1126/science.149.3688.1111. [DOI] [PubMed] [Google Scholar]
  3. BAEUMER A., PAU H., CONRADS H. [Demonstration of resochin or resochin derivatives in tissue]. Z Rheumaforsch. 1959 Dec;18:433–440. [PubMed] [Google Scholar]
  4. BRANDES D., BUETOW D. E., BERTINI F., MALKOFF D. B. ROLE OF LYSOSOMES IN CELLULAR LYTIC PROCESSES. I. EFFECT OF CARBON STARVATION IN EUGLENA GRACILIS. Exp Mol Pathol. 1964 Dec;90:583–609. doi: 10.1016/0014-4800(64)90036-x. [DOI] [PubMed] [Google Scholar]
  5. COHN Z. A., BENSON B. THE DIFFERENTIATION OF MONONUCLEAR PHAGOCYTES. MORPHOLOGY, CYTOCHEMISTRY, AND BIOCHEMISTRY. J Exp Med. 1965 Jan 1;121:153–170. doi: 10.1084/jem.121.1.153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. COHN Z. A., BENSON B. THE IN VITRO DIFFERENTIATION OF MONONUCLEAR PHAGOCYTES. II. THE INFLUENCE OF SERUM ON GRANULE FORMATION, HYDROLASE PRODUCTION, AND PINOCYTOSIS. J Exp Med. 1965 May 1;121:835–848. doi: 10.1084/jem.121.5.835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ciak J., Hahn F. E. Chloroquine: mode of action. Science. 1966 Jan 21;151(3708):347–349. doi: 10.1126/science.151.3708.347. [DOI] [PubMed] [Google Scholar]
  8. Cohen S. N., Yielding K. L. Inhibition of DNA and RNA polymerase reactions by chloroquine. Proc Natl Acad Sci U S A. 1965 Aug;54(2):521–527. doi: 10.1073/pnas.54.2.521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cohn Z. A., Fedorko M. E., Hirsch J. G. The in vitro differentiation of mononuclear phagocytes. V. The formation of macrophage lysosomes. J Exp Med. 1966 Apr 1;123(4):757–766. doi: 10.1084/jem.123.4.757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cohn Z. A., Hirsch J. G., Fedorko M. E. The in vitro differentiation of mononuclear phagocytes. IV. The ultrastructure of macrophage differentiation in the peritoneal cavity and in culture. J Exp Med. 1966 Apr 1;123(4):747–756. doi: 10.1084/jem.123.4.747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Cohn Z. A., Parks E. The regulation of pinocytosis in mouse macrophages. 3. The induction of vesicle formation by nucleosides and nucleotides. J Exp Med. 1967 Mar 1;125(3):457–466. doi: 10.1084/jem.125.3.457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Cohn Z. A., Parks E. The regulation of pinocytosis in mouse macrophages. II. Factors inducing vesicle formation. J Exp Med. 1967 Feb 1;125(2):213–232. doi: 10.1084/jem.125.2.213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Cohn Z. A. The regulation of pinocytosis in mouse macrophages. I. Metabolic requirements as defined by the use of inhibitors. J Exp Med. 1966 Oct 1;124(4):557–571. doi: 10.1084/jem.124.4.557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. De Duve C., Wattiaux R. Functions of lysosomes. Annu Rev Physiol. 1966;28:435–492. doi: 10.1146/annurev.ph.28.030166.002251. [DOI] [PubMed] [Google Scholar]
  15. Fedorko M. E., Hirsch J. G., Cohn Z. A. Autophagic vacuoles produced in vitro. I. Studies on cultured macrophages exposed to chloroquine. J Cell Biol. 1968 Aug;38(2):377–391. doi: 10.1083/jcb.38.2.377. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. HIRSCH J. G. Cinemicrophotographic observations on granule lysis in polymorphonuclear leucocytes during phagocytosis. J Exp Med. 1962 Dec 1;116:827–834. doi: 10.1084/jem.116.6.827. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Holtzman E., Novikoff A. B., Villaverde H. Lysosomes and GERL in normal and chromatolytic neurons of the rat ganglion nodosum. J Cell Biol. 1967 May;33(2):419–435. doi: 10.1083/jcb.33.2.419. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Morgan W. S., Fernando J., Alousi M. A. Studies of the biological activity of neutral red. Exp Mol Pathol. 1966 Oct;5(5):491–503. doi: 10.1016/0014-4800(66)90028-1. [DOI] [PubMed] [Google Scholar]
  19. NOVIKOFF A. B., ESSNER E., QUINTANA N. GOLGI APPARATUS AND LYSOSOMES. Fed Proc. 1964 Sep-Oct;23:1010–1022. [PubMed] [Google Scholar]
  20. 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]

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