Expression of DSD-1-PG in primary neural and glial-derived cell line cultures, upregulation by TGF-beta, and implications for cell-substrate interactions of the glial cell line Oli-neu.
DSD-1-PG is a chondroitin sulfate proteoglycan with neurite-outgrowth promoting properties expressed during development and upon lesion of neural tissues which has been defined with the specific monoclonal antibody 473HD. Double immunofluorescence studies performed on primary cerebellar cultures document that the proteoglycan is expressed on the surface of immature glial cells and the neural cell line Oli-neu, a model of mouse oligodendrocyte progenitors. Biochemical and immunoprecipitation studies performed with biosynthetically labelled Oli-neu and primary neural cells demonstrated that DSD-1-PG is expressed in vitro as a proteoglycan of 1000 kD apparent Mr with two core glycoproteins of 250 kD and 400 kD. In order to study the regulation of DSD-1-PG expression, an in vitro enzyme-linked immunosorbent assay based on Oli-neu and mAb 473HD was established. TGF-beta1-3 induced up-regulation of the proteoglycan, while various growth factors and cytokines did not significantly affect DSD-1-PG expression in both the supernatant and the extract of the culture monolayer. FACSCAN analysis suggested that the proteoglycan is upregulated on the surface of Oli-neu. Cell substrate adhesion assays revealed that this enhanced expression correlates with a selective reduction of adhesion to laminin, but not fibronectin or merosin, which could specifically be neutralized by antibodies to DSD-1-PG. We conclude that the proteoglycan contributes to the regulation of glial precursor interactions with the extracellular matrix.
Schnädelbach
O
O
Department of Neurobiology, University of Heidelberg, Germany.
Mandl
C
C
Faissner
A
A
D053674:Aggrecans; D000818:Animals; D002452:Cell Count / Q000187:drug effects; D018929:Cell Culture Techniques; D002460:Cell Line; D002531:Cerebellum / Q000166:cytology; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins / Q000096:biosynthesis* / Q000302:isolation & purification; D037181:Lectins, C-Type; D051379:Mice; D009457:Neuroglia / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D009836:Oligodendroglia / Q000378:metabolism; D011509:Proteoglycans; D016212:Transforming Growth Factor beta / Q000494:pharmacology*; D015854:Up-Regulation / Q000187:drug effects*
C508320:Acan protein, mouse; D053674:Aggrecans; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins; D037181:Lectins, C-Type; D011509:Proteoglycans; D016212:Transforming Growth Factor beta