Extracellular matrix of cultured glial cells: Selective expression of chondroitin 4-sulfate by type-2 astrocytes and their progenitors
- Istituto Superiore di Sanita', Rome (Italy)
We have studied the extracellular matrix composition of cultured glial cells by immunocytochemistry with different monoclonal and polyclonal antibodies. Double immunofluorescence experiments and metabolic labeling with (3H)glucosamine performed in different types of cerebellar and cortical cultures showed that bipotential progenitors for type-2 astrocytes and for oligodendrocytes synthesize chondroitin sulfate (CS) and deposit this proteoglycan in their extracellular matrix. The distribution of the various (3H)glucosamine-labeled glycosaminoglycans between the intracellular and the extracellular space was different. CS was present both within the cells and in the culture medium, although in different amounts. Bi-potential progenitors became also O4-positive during their development in vitro. At the stage of O4-positivity they were still stained with antibodies against CS. However, when the progenitor cells were maintained in serum-free medium and differentiated into Gal-C-positive oligodendrocytes, they became CS-negative. In the presence of fetal calf serum in the culture medium, the bipotential progenitors differentiated into GFAP-positive type-2 astrocytes. These cells still expressed CS: their Golgi area and their surface were stained with anti-CS antibodies. Staining with monoclonal antibodies specific for different types of CS (4-sulfate, 6-sulfate, and unsulfated) revealed that both bipotential progenitors and type-2 astrocytes synthesized only chondroitin 4-sulfate. Type-1 astrocytes were negative for both the polyclonal and the monoclonal anti-CS antibodies. Finally, type-2 astrocytes and their progenitors were weakly stained with anti-laminin antibodies and unstained with anti-fibronectin. Type-1 astrocytes were positive for both anti-laminin and anti-fibronectin antibodies and appeared to secrete fibronectin in the extracellular space.
- OSTI ID:
- 6828588
- Journal Information:
- Experimental Cell Research; (USA), Vol. 187:2; ISSN 0014-4827
- Country of Publication:
- United States
- Language:
- English
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EXTRACELLULAR SPACE
CHEMICAL COMPOSITION
NERVE CELLS
CELL DIFFERENTIATION
ANIMAL CELLS
CELL CULTURES
CEREBELLUM
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CHONDROITIN
FLUORESCENCE
GLUCOSAMINE
MONOCLONAL ANTIBODIES
RATS
TRACER TECHNIQUES
TRITIUM COMPOUNDS
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ANIMALS
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CARBOHYDRATES
CENTRAL NERVOUS SYSTEM
CEREBRUM
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LUMINESCENCE
MAMMALS
MONOSACCHARIDES
MUCOPOLYSACCHARIDES
NERVOUS SYSTEM
ORGANIC COMPOUNDS
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SACCHARIDES
SOMATIC CELLS
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550201* - Biochemistry- Tracer Techniques