Lipopolysaccharide (LPS) is one of the potent activators of macrophages. LPS was shown to induce cell spreading and large vacuoles in the cytoplasm of a macrophage-like cell line, JY3. These vacuoles were negative for acid phosphatase histochemistry and did not take up Lucifer yellow added to the medium. Latex beads were incorporated into cytoplasmic vesicles distinct from the vacuoles. These results indicated that the vacuoles are neither phagosomes nor lysosomes.
DiIC18(3), a specific marker of endoplasmic reticulum (ER), stained the vacuoles intensely, and DiOC6(3) stained the vacuoles at a density similar to nuclear envelope, suggesting ER origin of their membrane. Glucose-6-phosphatase, however, was not detected histochemically.
Vacuoles were also stained with wheat germ agglutinin-horseradish peroxidase (WGA-HRP) or WGA-biotin, suggesting that the vacuoles originated from plasma membrane-endosome-trans Golgi network-secretory granule pathway. Golgi markers, TPPase or BODIPY-ceramide were not localized to the vacuolar membrane.
These results indicate that the vacuoles may have dual origins; ER and plasma membrane-derived organelles.
References
1.
Tsan M., Berlin RDEffect of phagocytosis on membrane transport of nonelectrolytes. J Exp Med1971; 134: 1016—1035.
2.
Nitulescu GB, Bazin H., Spiegelberg HLSpecificity of Fc receptors for IgG2a, IgG1/IgG2b, and IgE on rat macrophages . J Exp Med1981; 154: 374—384.
3.
Knight KR, Vairo G., Hamilton JARegulation of pinocytosis in murine macrophages by colony stimulating factors and other agents. J Leukoc Biol1992; 51: 350—359.
4.
Hoffman RA, Langrehr JM, Dull KE, McCarthy SA, Jordan ML, Simmons RLMacrophage synthesis of nitric oxide in the mouse mixed leucocyte reaction . Transpl Immunol1994; 2: 313—320.
5.
Geng Y., Zhang B., Lotz M.Protein tyrosine kinase activation is required for lipopolysaccharide induction of cytokines in human blood monocytes. J Immunol1993; 151: 6692—6700.
6.
Abu-Lawi KI, Sultzer BMDiffering signal requirements for the activation of macrophages from C3H/HeJ and C3H/OuJ mice. FEMS Immunol Med Microbiol1994; 10: 31—38.
7.
Pawlowski NA , Kaplan G., Hamill AL, Cohn ZA, Scott WAArachidonic acid metabolism by human monocytes . J Exp Med1983; 158: 393—412.
8.
Vodovotz Y., Rossell D., Xie Q-W., Bogdan C., Nathan C.Vesicle membrane association of nitric oxide synthase in primary mouse macrophages . J Immunol1995; 154: 2914—2925.
9.
Kirikae T., Kirikae F., Oghiso Y., Nakano M.Microtubuledisrupting agents inhibit nitric oxide production in murine peritoneal macrophages stimulated with lipopolysaccharide or paclitaxel (taxol). Infect Immun1996; 64: 3379—3384.
10.
Dong Z., Qi X., Xie K., Fidler IJProtein tyrosine kinase inhibitors decrease induction of nitric oxide synthase activity in lipopolysaccharide-responsive and lipopolysaccharide-nonresponsive murine macrophages. J Immunol1993; 151: 2717—2724.
11.
Johnston RB , Kitagawa S.Molecular basis for the enhanced respiratory burst of activated macrophages. Fed Proc1985 ; 44: 2927—2932.
12.
Van Kessel KPM , Visser MR, Van Strijp JAG , VanKats-Renaud JH, Verhoef J. Cytotoxicity by human adherent cells: oxygen-dependent and -independent cytotoxic reactions by different cell populations. Immunology1986; 58: 291—296.
13.
Tracey KJ, Beutler B., Lowry SF et al. Shock and tissue injury induced by recombinant human cachectin . Science1986; 234: 470—474.
14.
Galanos C., Freudenberg MA, Reutter W.Galactosamine-induced sensitization to the lethal effects of endotoxin. Proc Natl Acad Sci USA1979; 76: 5939—5943.
15.
Hishinuma I. , Nagakawa J., Hirota K. et al. Involvement of tumor necrosis factor-α in development of hepatic injury in galactosamine-sensitized mice. Hepatology1990; 12: 1187—1191.
16.
Nagakawa J. , Hishinuma I., Hirota K. et al. Interleukin-1α enhances hepatotoxicity of tumor necrosis factor-α in galactosamine-sensitized mice. Immunopharmacol Immunotoxicol1991; 13: 485—498.
17.
Auger MJ, Ross JABiology of the macrophage. In: Lewis CE, McGee JO'D, eds. The Macrophage. New York: IRL Press1992; 39—41.
18.
Amano F., Akamatsu Y.A lipopolysaccharide (LPS)-resistant mutant isolated from a macrophage like cell line, J774.1, exhibits an altered activated-macrophage phenotype in response to LPS. Infect Immun1991; 59: 2166—2174.
19.
Hatcher GE, Lambrecht RSAugumentation of macrophage phagocytic activity by cell-free extracts of selected lactic acid-producing bacteria. J Dairy Sci1993; 76: 2485—2492.
20.
Sawada H., Esaki M.Use of nanogold followed by silver enhancement and gold toning for postembedding immunolocalization in osmium-fixed, Epon-embedded tissues. J Electron Microsc (Tokyo)1994; 43:361—366.
21.
Murakami T. , Ishikawa H., Peachey LDConfocal microscopic 3D visualization of membranous organelles in living cultured fibroblasts. Bioimage1994; 2: 21—28.
22.
Terasaki M. , Song J., Wong JR, Weiss MJ, Chen LBLocalization of endoplasmic reticulum in living and glutaraldehyde-fixed cells with fluorescent dyes. Cell1984; 38: 101—108.
23.
Baumann O., Kitazawa T., Somlyo APLaser confocal scanning microscopy of the surface membrane/T-tubular system and the sarcoplasmic reticulum in insect striated muscle stained with DiIC18(3) . J Struct Biol1990; 105: 154—161.
24.
Terasaki M. , Jaffe LAOrganization of the sea urchin egg endoplasmic reticulum and its reorganization at fertilization. J Cell Biol1991; 114: 929—940.
25.
Pagano RE, Martin OC, Kang HC, Haugland RPA novel fluorescent ceramide analogue for studying membrane traffic in animal cells: accumulation at the Golgi apparatus results in altered spectral properties of the sphingolipid precursor. J Cell Biol1991; 113: 1267—1279.
26.
Barka T., Anderson PJHistochemical methods for acid phosphatase using hexazonium pararoseanilin as coupler. J Histochem Cytochem1962; 10: 741—753.
27.
Saito T.Ultracytochemistry of enzymes in cell biology of the retina. J Electron Microsc (Tokyo) 1991; 40: 312—318.
28.
Wachstein M. , Meisel E.On the histochemical demonstration of glucose-6-phosphatase . J Histochem Cytochem1956; 4: 592.
29.
Novikoff AB , Goldfischer S.Nucleosidediphosphatase activity in the Golgi apparatus and its usefulness for cytological studies. Proc Natl Acad Sci USA1961; 47: 802—810.
30.
Tartakoff AMPerturbation of vesicular traffic with the carboxylic ionophore monensin. Cell1993; 32: 1026—1028.
31.
Griffiths G. , Quinn P., Warren G.Dissection of the Golgi complex. I. Monensin inhibits the transport of viral membrane proteins from medial to trans Golgi cisternae in baby hamster kidney cells infected with Semliki Forest virus . J Cell Biol1983; 96: 835—850.
32.
Klausner RD , Donaldson JG, Lippincott-Schwarts J.Brefeldin A: insights into the control of membrane traffic and organelle structure. J Cell Biol1992; 116: 1071—1080.
33.
Fujiwara T. , Oda K., Yokota S., Takatsuki A., Ikehara Y.Brefeldin A causes disassembly of the Golgi complex and accumulation of secretory proteins in the endoplasmic reticulum . J Biol Chem1988; 263: 18545—18552.
34.
Hendricks LC , McClanahan SL, Palade GE, Farquhar MGBrefeldin A affects early events but does not affect late events along the exocytic pathway in pancreatic acinar cells . Proc Natl Acad Sci USA1992; 89: 7242—7246.
35.
Hotchkiss RDA microchemical reaction resulting in the staining of polysaccharide structures in fixed tissue preparations. Arch Biochem1948; 16: 131—141.
36.
Daddi L.Nouvelle methode pour colorer la graisse dans les tissus. Arch Ital Biol1896; 26: 143—146.
37.
Smith BM, Sturm RJ, Carchman RACalcium modulation of phorbol ester-induced alteration in murine macrophage morphology. Cancer Res1983; 43: 3385—3391.
38.
Cheung HT, Cantarow WD, Sundharadas G.Colchicine and cytochalasin B (CB) effects on random movement, spreading and adhesion of mouse macrophages. Exp Cell Res1978; 111: 95—103.
39.
Montgomery RR , Webster P., Mellman I.Accumulation of indigestible substances reduces fusion competence of macrophage lysosomes. J Immunol1991; 147: 3087—3095.
40.
Iida H., Shibata Y.Delivery of lectin-labeled membrane to the trans-Golgi network and secretory granules in cultured atrial myocytes. J Histochem Cytochem1989; 37: 1885—1892.
41.
Raub TJ, Koroly Mjo, Roberts RMRapid endocytosis and recycling of wheat germ agglutinin binding sites on CHO cells: Evidence for two compartments in a nondegradative pathway. J Cell Physiol1990; 144: 52—61.
42.
Raub TJ, Koroly Mjo, Roberts RMEndocytosis of wheat germ agglutinin binding sites from the cell surface into a tubular endosomal network. J Cell Physiol1990; 143: 1—12.
43.
Stieber A., Gonatas JO, Gonatas NKDifferences between the endocytosis of horseradish peroxidase and its conjugate with wheat germ agglutinin by cultured fibroblasts. J Cell Physiol1984; 119: 71—76.
Gonatas NK, Stieber A., Hickey WF, Herbert SH, Gonatas JOEndosomes and Golgi vesicles in adsorptive and fluid phase endocytosis. J Cell Biol1984; 99: 1379—1390.
46.
Boocock CA, Jones GEColony-stimulating factor-1 induces rapid behavioural responses in the mouse macrophage cell line, BAC1.2F5. J Cell Sci1989; 93: 447—456.
47.
Racoosin EL , Swanson JAM-CSF-induced macropinocytosis increase solute endocytosis but not receptor-mediated endocytosis in mouse macrophages. J Cell Sci1992; 102: 867—880.
48.
Swanson JAPhorbol esters stimulate macropinocytosis and solute flow through macrophages . J Cell Sci1989; 94: 135—142.
49.
Luchi M., Munford RSBinding, internalization, and deacylation of bacterial lipopolysaccharide by human neutrophils. J Immunol1993; 151: 959—969.
