Phorbol myristate acetate stimulates phagosome-lysosome fusion in mouse macrophages

Margaret Kielian, Z. A. Cohn

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

The effect of the tumor promoter phorbol myristate acetate (PMA) on phagosome-lysosome (P-L) fusion in mouse macrophages has been studied using a previously described (10) r286w7 =cycloheximide fluorescence assay. Treatment with 01-1.0 μg PMA/ml caused a striking increase in the rate and extent of P-L fusion. Exposure of cells to phorbol, free myristate, or the monoesters of PMA did not reproduce this effect. Macrophages required from 2 to 3 h of pretreatment to express maximal P-L fusion, and this was maintained for at least 20 h when cells were returned to PMA-free medium. Catalase, superoxide dismutase, indomethacin, and hydrocortisone, agents that are known to block the effect of PMA on H2O2, O-2, prostaglandins, or plasminogen activator, did not affect the stimulation of P-L fusion by PMA. The protein-synthesis inhibitors puromycin and chcloheximide did block the PMA effect under conditions in which the high fusion rate of 4-d cells was not affected. Labeled PMA was rapidly taken up by macrophages, with a plateau of uptake at ~3 h. When cells were returned to PMA-free medium, cell-associated label was rapidly released, returning to background levels within 1 h. The released label was found to be a metabolite of PMA by thin-layer chromatography. This product migrated between the monoester phorbol-12-myristate and free phorbol. Rapid metabolism of PMA was also observed by a macrophage cell line, J774, and, to a lesser extent, by primary rat embryo fibroblasts.

Original languageEnglish (US)
Pages (from-to)101-111
Number of pages11
JournalJournal of Experimental Medicine
Volume154
Issue number1
StatePublished - 1981
Externally publishedYes

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Phagosomes
Tetradecanoylphorbol Acetate
Lysosomes
Macrophages
Puromycin
Protein Synthesis Inhibitors
Plasminogen Activators
Myristic Acid
Cycloheximide
Thin Layer Chromatography
Indomethacin
Carcinogens
Catalase
Superoxide Dismutase
Prostaglandins
Hydrocortisone
Embryonic Structures
Fibroblasts
Fluorescence

ASJC Scopus subject areas

  • Immunology

Cite this

Phorbol myristate acetate stimulates phagosome-lysosome fusion in mouse macrophages. / Kielian, Margaret; Cohn, Z. A.

In: Journal of Experimental Medicine, Vol. 154, No. 1, 1981, p. 101-111.

Research output: Contribution to journalArticle

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