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)