The dedifferentiation of chondrocytes in culture is classically associated with a transition from a rounded to a spread morphology. However, the loss of chondroitin sulfate proteoglycan (CSPG) and type II collagen gene expression (markers of the differentiated chondrocyte) does not occur for all polygonal or fibroblast-like cells at the same stage of culture. Furthermore, it has been demonstrated that retinoic acid-dedifferentiated chondrocytes can reexpress type II collagen if treated by the microfilament disruptive drug dihydrocytochalasin B, without a return to the spherical shape. In the present study, we have investigated by fluorescent double-staining whether the synthesis of both CSPG and type II collagen by dedifferentiating chick chondrocytes in low density cultures is dependent on a type of actin organization. We report that the synthesis of CSPG and type II collagen synthesis is coincident with the presence of a faint microfibrillar actin architecture but is absent in chondrocytes showing well defined actin cables. This correlation was observed independently of the shapes exhibited by the cells. Moreover, type I collagen (marker of the dedifferentiated chondrocyte) is synthesized mainly in cells showing large actin cables. This study, performed in the absence of drugs, suggests that actin organization, rather than changes in cell shape, is involved in modulating the chondrogenic phenotype in vitro.The dedifferentiation of chondrocytes in culture is classically associated with a transition from a rounded to a spread morphology. However, the loss of chondroitin sulfate proteoglycan (CSPG) and type II collagen gene expression (markers of the differentiated chondrocyte) does not occur for all polygonal or fibroblast-like cells at the same stage of culture. Furthermore, it has been demonstrated that retinoic acid-dedifferentiated chondrocytes can reexpress type II collagen if treated by the microfilament disruptive drug dihydrocytochalasin B, without a return to the spherical shape. In the present study, we have investigated by fluorescent double-staining whether the synthesis of both CSPG and type II collagen by dedifferentiating chick chondrocytes in low density cultures is dependent on a type of actin organization. We report that the synthesis of CSPG and type II collagen synthesis is coincident with the presence of a faint microfibrillar actin architecture but is absent in chondrocytes showing well defined actin cables. This correlation was observed independently of the shapes exhibited by the cells. Moreover, type I collagen (marker of the dedifferentiated chondrocyte) is synthesized mainly in cells showing large actin cables. This study, performed in the absence of drugs, suggests that actin organization, rather than changes in cell shape, is involved in modulating the chondrogenic phenotype in vitro.