Expression of fibronectin and laminin by different types of mouse glial cells cultured in a serum-free medium.
The expression of fibronectin and laminin by cultured glial cells was studied. The glial culture from neonatal mouse cerebra maintained in a chemically defined, serum-free medium consisted of type-1 astrocytes, oligodendrocyte-type-2 astrocyte (O-2A) progenitor cells, oligodendrocytes and type-2 astrocytes. Double-labelling immunofluorescent experiments performed using the mixed glial culture indicated that fibronectin and laminin are expressed in different patterns among the glial subtypes. The staining intensities with anti-fibronectin or anti-laminin antibodies decreased in the order: type-1 astrocytes, O-2A progenitor cells and type-2 astrocytes. Both molecules were deposited in a fibrillar matrix underneath type-1 astrocytes, whereas only intracytoplasmic localization of these molecules was observed with O-2A progenitor cells and type-2 astrocytes. Western blot analysis showed that glial fibronectin has a slightly higher molecular weight than mouse plasma fibronectin (230 kDa) and that glial laminin is a variant with a 220 kDa B chain present and the 400 kDa A chain missing. Using enzyme-linked immunosorbent assays (ELISA), these molecules were detected in the glial extracellular matrix at the concentration of 4 ng/10(6) cells. A large amount of fibronectin (82 ng/10(6) cells) was secreted into the culture medium, while secretion of laminin was not detected.
Institute for Fermentation, (IFO), Osaka, Japan.
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T
Yoshida
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Takeuchi
D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D004797:Enzyme-Linked Immunosorbent Assay; D005353:Fibronectins / Q000096:biosynthesis* / Q000235:genetics; D005455:Fluorescent Antibody Technique; D007797:Laminin / Q000096:biosynthesis* / Q000235:genetics; D051379:Mice; D008856:Microscopy, Fluorescence; D009457:Neuroglia / Q000166:cytology / Q000378:metabolism*
D016895:Culture Media, Serum-Free; D005353:Fibronectins; D007797:Laminin