Basic fibroblast growth factor (bFGF) stimulates proliferation of chondrocytes and their extracellular matrix synthesis but inhibits terminal differentiation to hypertrophic cells (Kato, Y., and Iwamoto, M., (1990) J. Biol. Chem. 265, 5903-5909). In the present study, we examined changes in bFGF binding during chondrocyte cytodifferentiation. In cultures of pelleted growth plate chondrocytes, binding of 125I-bFGF to 140-kDa receptors was observed during the mitotic and matrix-forming stages but decreased to a very low level as chondrocytes became hypertrophic. Scatchard plot analysis showed that the decrease in binding of bFGF was due to a decrease in the number not in the affinity of the receptor. The loss of bFGF receptor was associated with a decrease in biological responses to bFGF. On the other hand, the binding of transforming growth factor-beta and epidermal growth factor was constant throughout all stages of growth plate chondrocytes. A rapid decrease in bFGF binding was not observed with articular chondrocytes or bFGF-exposed growth plate chondrocytes, perhaps because they scarcely underwent terminal differentiation. A decrease in bFGF binding associated with terminal differentiation in situ was also demonstrated by examination of sequential slices of growth plates. These observations suggest that rapid reduction in bFGF receptor is a special event during terminal differentiation.