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J Exp Med. 1966 October 1; 124(4): 557–571.
PMCID: PMC2138244

THE REGULATION OF PINOCYTOSIS IN MOUSE MACROPHAGES

I. METABOLIC REQUIREMENTS AS DEFINED BY THE USE OF INHIBITORS

Abstract

A method is described to study the formation of pinocytic vesicles in cultivated mouse macrophages. Vesicles which arise in the peripheral cytoplasm and are in transit to the centrosphere region are enumerated by the phase-contrast microscopy of glutaraldehyde-fixed cells. Under these conditions there is a prompt reversible response of vesicle formation to calf serum factors in the external environment. The reduction of pinosome formation by a variety of metabolic inhibitors was then studied in a medium containing 50% newborn calf serum. Inhibitors of both glycolysis and respiration reduced vesicle formation to low levels. This influence was most striking with cyanide, antimycin A, and anaerobiosis. This indicates an important contribution of an intact respiratory pathway in pinocytosis. Both 2,4-dinitrophenol and oligomycin inhibited vesicle formation at low concentrations. These data suggest a central role of ATP as an energy source for vesicle formation. Inhibitors of protein synthesis, such as puromycin and p-fluorophenylalanine, produced a prompt reduction in vesicle formation. The action of p-fluorophenylalanine was effectively reversed with L-phenylalanine. Actinomycin D depressed pinocytosis to low levels at concentrations of 0.1 to 0.003 µg/ml. This effect was observed only after a 120 min lag phase. A 10°C reduction in ambient temperature lowered vesicle counts to 30% of control preparations at 37°C.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • HOLTER H. Pinocytosis. Int Rev Cytol. 1959;8:481–504. [PubMed]
  • HOLTZER H, HOLTZER S. The in vitro uptake of fluorescein labelled plasma proteins. I. Mature cells. C R Trav Lab Carlsberg. 1960;31:373–408. [PubMed]
  • RYSER HJ. THE MEASUREMENT OF I131-SERUM ALBUMIN UPTAKE BY TUMOR CELLS IN TISSUE CULTURE. Lab Invest. 1963 Oct;12:1009–1017. [PubMed]
  • ROSE GG. Microkinetospheres and VP satellites of pinocytic cells observed in tissue cultures of Gey's strain HeLa with phase contrast cinematographic techniques. J Biophys Biochem Cytol. 1957 Sep 25;3(5):697–704. [PMC free article] [PubMed]
  • BARRNETT RJ, BALL EG. Metabolic and ultrastructural changes induced in adipose tissue by insulin. J Biophys Biochem Cytol. 1960 Sep;8:83–101. [PMC free article] [PubMed]
  • Cohn ZA, Hirsch JG, Fedorko ME. 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. [PMC free article] [PubMed]
  • Cohn ZA, Fedorko ME, Hirsch JG. The in vitro differentiation of mononuclear phagocytes. V. The formation of macrophage lysosomes. J Exp Med. 1966 Apr 1;123(4):757–766. [PMC free article] [PubMed]
  • COHN ZA, 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. [PMC free article] [PubMed]
  • SBARRA AJ, KARNOVSKY ML. The biochemical basis of phagocytosis. I. Metabolic changes during the ingestion of particles by polymorphonuclear leukocytes. J Biol Chem. 1959 Jun;234(6):1355–1362. [PubMed]
  • COHN ZA, MORSE SI. Functional and metabolic properties of polymorphonuclear leucocytes. I. Observations on the requirements and consequences of particle ingestion. J Exp Med. 1960 May 1;111:667–687. [PMC free article] [PubMed]
  • HUIJING F, SLATER EC. The use of oligomycin as an inhibitor of oxidative phosphorylation. J Biochem. 1961 Jun;49:493–501. [PubMed]
  • COHN ZA, BENSON B. THE IN VITRO DIFFERENTIATION OF MONONUCLEAR PHAGOCYTES. I. THE INFLUENCE OF INHIBITORS AND THE RESULTS OF AUTORADIOGRAPHY. J Exp Med. 1965 Feb 1;121:279–288. [PMC free article] [PubMed]

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