Characterization and immunocytochemical localization of actin and fibronectin in haemocytes of the musselMytilus galloprovincialis
Tóm tắt
Cell-extracellular matrix interactions are recognized to be important for human leucocyte functions, including chemotaxis and phagocytosis. These activities depend on a reorganization of the microfilament actin (F-actin) promoted by fibronectin, one of the major components of extracellular matrices. Although invertebrate haemocytes are, in many aspects, similar to the human granulocyte-monocyte-macrophage cell lineage, actin and fibronectin have not been well studied in these cells. Consequently, the characterization and structural organization of actin and fibronectin in mussel (Mytilus galloprovincialis) haemocytes was investigated using Western blotting analysis, indirect immunofluorescence and immunoelectron microscopy. Actin was immunocharacterized by an anti-total actin monoclonal antibody. Fibronectin was immunocharacterized by an autologous polyclonal antiserum directed against the protein of mussel haemolymph. Actin was mainly localized along the peripheral cytoplasm of the haemocyte. The distribution of the F-actin microfilaments was assayed with Rhodamine-labelled phalloidin. F-actin was associated mainly with stress-fibres of spreading haemocytes and with microspikes at the adhesion sites. The labelling by the anti-fibronectin antiserum of the haemocyte rough endoplasmic reticulum vesiles, revealed by immunoelectron microscopy, suggests that these cells are involved in fibronectin biosynthesis. Gold particles were also present along the outer surfaces of the cell plasma membrane and its protrusions. Mussel fibronectin was localized immunohistochemically at the adhesion sites and in the extracellular matrix fibrils. The relationships between fibronectin and the actin cystoskeleton inMytilus galloprovincialis haemocytes are discussed.
Tài liệu tham khảo
Alberts, B. A., Bray, D., Lewis, J., Raff, M., Roberts, K. &Watson, J. D. (1994)Molecular Biology of the Cell, New York, London: Garland Publishing.
Alvarez, M. R., Friedl, F. E., Johnson, J. S. &Hinsch, G. W. (1989) Factors affectingin vitro phagocytosis in oyster hemocytes.J. Invert. Pathol. 54, 233–41.
Ball, E. E., de Couet, H. G., Horn, P. L. &Quinn, J. M. A. (1987) Haemocytes secrete basement membrane components in embryonic locusts.Development 99, 255–9.
Bayne, C. J. (1983) Molluscan immunobiology. InThe mollusca (edited by A. S. M. Salleuddin & K. M., Wilbur), Vol 5:Physiology, Part 2, pp. 407–86. London: Academic Press.
Bengtsson, T., Jaconi, M. E. E., Gustafson, M., Magnusson, K.-E. Theler, J.-M., Lew, D. P. &Stendahl, O. (1993) Actin dynamics in human neutrophils during adhesion and phagocytosis is controlled by changes in intracellular free calcium.Eur. J. Cell Biol. 62, 49–59.
Blystone, S. D., Graham, I. L., Lindberg, F. P. &Brown, E. (1994) Integrin αvβ3 differentially regulates adhesive and phagocytic functions of the fibronectin receptor α5β1.J. Cell Biol. 127, 1129–37.
Bradford, M. M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.Anal. Biochem. 72, 248–54.
Cheng, C. T. &Combes, C. (1990) Influence of environmental factors on the invasion of molluscs by parasites: with special reference to Europe. InBiological Invasions in Europe and the Mediterranean Basin (edited by F. Di Castri, A. J. Hanse, and M. Debussche) pp. 307–32. Dordrecht: Kluwer Academic.
Cheng, T. C. &Howland, K. H. (1982) Effect of colchicine and cytochalasin B on chemotaxis of oyster (Crassostrea virginica) hemocytes.J. Invert. Pathol. 40, 150–2.
Cooper, E. L. (1992) Overview of immunoevolution.Boll. Zool. 59, 119–28.
Di Rosa, I., Contenti, S., Fagotti, A., Simoncelli, F., Principato, G. B., Panara, F. &Pascolini, R. (1994). Fibronectin from the hemolymph of the marine bivalveMytilus galloprovincialis. Purification, immunological characterization and immunocytochemical localization.Comp. Biochem. Physiol. 107B, 625–32.
Doherty, D. E., Henson, P. M. &Clark, R. A. F. (1990) Fibronectin fragments containing the RGDS cell-binding domain mediate monocyte migration into the rabbit lung: a potential mechanism for C5 fragment-induced monocyte lung accumulation.J. Clin. Invest. 86, 1065–75.
Fernandez-Segura, E., Garcia, J. M., Santos, J. L. &Campos, A. (1995) Shape, F-actin, and surface morphology changes during chemotactic peptide-induced polarity in human neutrophils.Anat. Rec. 241, 519–28.
Genedani, S., Bernardi, M., Ottaviani, E., Franceschi, C., Leung, M. K. &Stefano, G. (1994) Differential modulation of invertebrate hemocyte motility by CRF, ACTH, and its fragments.Peptides 15, 203–6.
Hansen, P. D. (1992) Phagocytosis inMytilus edulis, a system for understanding the sublethal effects of anthropogenic pollutants and the use of AOX as an integrating parameter for the study of equilibria between chlorinated organics inDreissena polymorpha following long term exposures. InLimnologie Aktuell, Vol. 4. (Edited by R. Neumann and P. Jenner), pp. 171–83. New York: Gustav Fischer Verlag.
Howard, T. H. &Oresajo, C. (1985) The kinetics of chemmotactic peptide induced change in F-actin content, F-actin distribution and shape of human neutrophils.J. Cell Biol. 101, 1078–85.
Hughes, T. K., Smith, F. M., Chin, R., Cadet, P., Sinisterra, J., Leung, M. K., Shipp, M. A., Scharrer, B. &Stefano, G. B. (1990) Interaction of immunoreactive monokines (interleukin 1 and tumor necrosis factor) in the bivalve molluscMytilus edulis.Proc. Natl Acad. Sci. USA 87, 4426–9.
Hughes, I. K., Smith, E. M., Barnett, J. A., Charles, R. &Stefano, G. B. (1991a) LPS stimulated invertebrate hemocytes: a role for immunoreactive TNF and IL-1.Dev. Comp. Immunol. 15, 117–22.
Hughes, T. K., Smith, E. M., Barnett, J. A., Charles, R. &Stefano, G. B. (1991b) Lipopolysaccharide and opioids activate distinct populations ofMytilus edulis immunocytes.Cell Tissue Res. 264, 317–20.
Hynes, R. O. (1990)Fibronectins. New York: Springer Verlag.
Keller, H. U. &Niggli, V. (1995) Cochicine-induced stimulation of PMN motility related to cytoskeletal changes in actin, α-actinin, and myosin.Cell Motil. Cytoskel. 25, 10–18.
Laemmli, U. K. (1990) Cleavage of structural proteins during the assembly of the head of bacteriophage T4.Nature 227, 680–5.
Lerat, H., Lissitzky, J. C., Singer, J. W., Keating, A., Herve, P. &Charbord, P. (1993) Role of stromal cells and macrophages in fibronectin biosynthesis and matrix assembly in human long-term marrow cultures.Blood 82, 1480–92.
Millar, D. A. &Ratcliffe, N. A. (1989) The evolution of blood cells: facts and enigmas.Endeavor 13, 72–7.
Moore, C. A., Beckmann, N. &Morse, M. P. (1992) Cytoskeletal structure of diseased and normal hemocytes ofMya arenaria.J. Invert. Pathol. 60, 141–7.
Noel, D., Bachere, E., &Mialhe, E. (1993) Phagocytosis associated chemiluminescence of hemocytes inMytilus edulis (Bivalvia),Dev. Comp. Immunol. 17, 483–93.
Pakianathan, D. R. (1995) Extracellular matrix proteins and leukocyte function.J. Leukocyte Biol. 57, 699–702.
Pascolini, R., Panara, F., Di Rosa I., Fagotti, A. &Lorvik, S. (1992) Characterization and fine-structural localization of actin-and fibronectin-like proteins in planaria (Dugesia luguris s.l.).Cell Tissue Res. 267, 499–506.
Pipe, R. K. (1990a) Differential binding of lectins to haemocytes of the musselMytilus edulis.Cell Tissue Res. 261, 261–8.
Pipe, R. K. (1990b) Hydrolytic enzymes associated with the granular haemocytes of the marine musselMytilus edulis.Histochem. J. 22, 595–603.
Pommier, C. G., Inada, S., Fries, L. F., Takahashi, T., Frank, M. M. &Brown, E. J. (1983) Plasma fibronectin enhances phagocytosis of opsonized particles by human peripheral blood monocytes.J. Exp. Med. 157, 1844–54.
Rewrantz, L., Daniels, J. &Hansen, P. D. (1985) Lectin-binding to hemocytes ofMytilus edulis.Dev. Comp. Immunol. 9, 203–10.
Ruoslathi, E. (1988) Fibronectin and its receptors.Ann. Rev. Biochem. 57, 375–413.
Scapigliati, G., Fausto, A. M. &Mazzini, M. (1993) Morphological and cytoskeletal characterization of haemocytes in stick insects (Phasmatodea).Boll. Zool. 60, 25–32.
Schneeweiß, H. &Rewrantz, L. (1993) Analysis of the attraction of haemocytes fromMytilus edulis by molcules of bacterial origin.Dev. Comp. Immunol. 17, 377–87.
Stefano, G. B., Cadet, P. &Scharrer, B. (1989a) Stimulatory effects of opioid neuropeptides on locomotory activity and conformational changes in invertebrate and human immunocytes: evidence for a new subtype of delta receptor.Proc. Natl Acad. Sci. USA 86, 6307–11.
Stefano, G. B., Leung, M. K., Zhao, X. &Scharrer, B. (1989b) Evidence for the involvement of opioid neuropeptides in the adherence and migration of immunocompetent invertebrate hemocytes.Proc. Natl Acad. Sci. USA 86, 626–30.
Stossel, T. P. (1989) From signal to pseudopod. How cells control cytoplasmic actin assembly.J. Biol. Chem. 264, 18261–4.
Stossel, T. P. (1992) The mechanical responses of white blood cells. InInflammation: Basic Principles and Clinical Correlates (edited by J. I. Gallin, I. M. Goldstein and R. Snyderman, 2nd edn, pp. 459–75. New York: Raven Press.
Suzuki, T. &Funakoshi, S. (1992) Isolation of a fibronectin-like molecule from a marine bivalve,Pinctada fucata, and its secretion by amebocytes.Zool. Sci. 9, 541–50.
Towbin, M., Staehelin, T. &Gordon, J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.Proc. Natl Acad. Sci. USA 76, 4350–4.
Van der Knaap, W. P. W., Adema, C. M. &Sminia, T. (1993) Invetebrate blood cells: morphological and functional aspects of the haemocytes in the pond snailLymnaea stagnalis.Comp. Haematol. Int. 3, 20–6.
Watts, R. G., Crispens, M. A. &Howard, T. H. (1991) A quantitative study of the role of F-actin in producing neutrophil shape.Cell Motil. Cytoskel. 19, 159–68.
Watts, R. G., Deaton, J. D. &Howard, T. H. (1995) Dynamics of Triton-insoluble and Triton-soluble F-actin pools in calcium-activated human polymorphonuclear leukocytes: evidence for regulation by gelsolin.Cell Motil. Cytoskel. 30, 136–45.
Yang, K. D., Augustine, N. H., Gonzales, L. A., Bohnsack, J. F. &Hill, H. R. (1988) Effects of fibronectin on the interaction of polymorphonuclear leukocytes with unopsonized and antibody-opsonized bacteria.J. Infect. Dis. 158, 823–30.
Yang, K. D., Bohnsack, J. F., Hawley, M. M., Egan, M. L., Pritchard, D. G. &Hill, H. R. (1990) Effect of fibronectin on IgA-mediated uptake of type III group B streptococci by phagocytes.J. Infect. Dis. 161, 236–41.
Yang, K. D., Augustine, N. H., Shaio, M.-F., Bohnsack, J. F. &Hill, H. R. (1994) Effects of fibronectin on actin organization and respiratory burst activity in neutrophils, monocytes, and macrophages.J. Cell Physiol. 158, 347–53.