Expression of fibronectin and laminin by different types of mouse glial cells cultured in a serum-free medium |
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Authors: | Touho Yoshida Masao Takeuchi |
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Affiliation: | (1) Institute for Fermentation, Osaka (IFO), 17-85, Juso-honmachi 2-chome, 532 Yodogawa-ku, Osaka, Japan |
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Abstract: | 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/106 cells. A large amount of fibronectin (82 ng/106 cells) was secreted into the culture medium, while secretion of laminin was not detected. |
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Keywords: | Extracellular matrix fibronectin glia laminin |
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