Spatial organization of the extracellular matrix modulates the expression of PDGF-receptor subunits in mesangial cells.
The aim of this study was to test the hypothesis that changes in the extracellular matrix environment regulate rat mesangial cell growth by modulation of the expression of both PDGF-receptor alpha- and beta-subunits. We investigated the mitogenic effects of the PDGF isoforms AA, AB and BB in conventional two-dimensional (2D) culture on laminin, fibronectin, type I, IV and V collagen and in the different spatial organization of matrix in type I collagen gels in three-dimensional culture (3D). In 2D culture PDGF BB was a potent mitogen, AB elicited an intermediate response while AA had no effect on cell proliferation. Extracellular matrix did not modify the PDGF responsiveness in 2D-culture. The different effects of the three PDGF isoforms were due to differential expression and isoform specific association of the PDGF-receptor subunits. Specifically, the beta-receptor was strongly expressed, whereas the alpha-receptor was only barely detectable on the cell surface. Metabolic labeling revealed synthesis and intracellular accumulation of the complete alpha-receptor protein, and treatment with suramin increased its surface expression, suggesting continuous receptor down-regulation by endogenous PDGF. Morphological and ultrastructural analysis in 3D culture revealed a change in mesangial cell phenotype, forming a branching network of multicellular structures. Assessment of proliferation in 3D culture showed quiescent cells and PDGF unresponsiveness. Investigation of the PDGF beta-receptors revealed a rapid down-regulation in 3D culture; both receptor subunits were not detectable on the cell surface. We conclude that 3D culture promotes the induction of a different mesangial cell phenotype.(ABSTRACT TRUNCATED AT 250 WORDS)
Department of Pathology, Yale University School of Medicine, New Haven, Connecticut.
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M
Marx
T O
TO
Daniel
M
M
Kashgarian
J A
JA
Madri
D000818:Animals; D000077214:Becaplermin; D002478:Cells, Cultured; D003584:Cytological Techniques; D015536:Down-Regulation; D005109:Extracellular Matrix / Q000648:ultrastructure*; D005920:Glomerular Mesangium / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D007536:Isomerism; D008934:Mitogens / Q000494:pharmacology; D010641:Phenotype; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D020574:Proto-Oncogene Proteins c-sis; D017479:Receptors, Platelet-Derived Growth Factor / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000494:pharmacology
D008934:Mitogens; D010982:Platelet-Derived Growth Factor; D020574:Proto-Oncogene Proteins c-sis; D011994:Recombinant Proteins; C075278:platelet-derived growth factor A; D000077214:Becaplermin; D017479:Receptors, Platelet-Derived Growth Factor