Characterization and immunocytochemical localization of actin and fibronectin in haemocytes of the mussel Mytilus galloprovincialis.
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 vesicles, 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 cytoskeleton in Mytilus galloprovincialis haemocytes are discussed.
Panara
F
F
Istituto di Anatomia Comparata, Università di Perugia, Italy.
Di Rosa
I
I
Fagotti
A
A
Simoncelli
F
F
Mangiabene
C
C
Pipe
R K
RK
Pascolini
R
R
D000199:Actins / Q000032:analysis*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D049872:Bivalvia / Q000737:chemistry*; D015153:Blotting, Western; D005353:Fibronectins / Q000032:analysis*; D019084:Fluorescent Antibody Technique, Indirect; D006434:Hemocytes / Q000737:chemistry* / Q000648:ultrastructure; D016253:Microscopy, Immunoelectron; D010590:Phalloidine
D000199:Actins; D000911:Antibodies, Monoclonal; D005353:Fibronectins; D010590:Phalloidine