In mammals and birds, most of the skeletal bones develop via endochondral ossification. Chondrocytes in the cartilaginous anlagen undergo processes of maturation such as hypertrophy, calcification and apoptosis. Concomitantly, osteoblasts are recruited to replace the cartilage scaffold gradually with bone matrix and become osteocytes in the trabecular bones. Throughout the successive development of bones, several gene products have been identified as being the components of the molecular mechanism regulating bone development. Transcription factor SOX9 plays essential roles during developmental steps from undifferentiated mesenchymal cells to proliferating chondrocytes, meanwhile, it inhibits transition of proliferating chondrocytes to hypertrophy. Other transcription factors RUNX2 and OSTERIX are critical in osteoblast differentiation, and RUNX2 is also essential for chondrocyte maturation such as hypertrophy and matrix mineralization. GDF5, a protein belonging to the transforming growth factor beta superfamily, is involved in joint formation and chondrogenesis. The limb skeleton of one of the ancestral tetrapod, anuran amphibians also develops through cartilaginous anlagen to bones, but their skeletogenesis has some unique characteristics compared with that of mammals and birds. Anuran amphibians develop and grow with less bone trabeculae and poor epiphyseal growth plates, and its endochondral ossification was found to be a delayed process. In order to address the characteristic skeletal development of anuran amphibians, we cloned Xenopus tropicalis RUNX2 (Xt-runx2), OSTERIX (Xt-osterix) and GDF5 (Xt-gdf5) homologue, and observed expression patterns together with Xt-sox9. In X. tropicalis limbs, histological observation and section in situ hybridization analysis suggest that Xt-SOX9 is involved in chondrogenesis, Xt-RUNX2 and Xt-OSTERIX are involved in osteogenesis, and Xt-GDF5 is involved in joint formation. In the cartilaginous anlagen, Xt-runx2 expression was found in perichondrium and immature chondrocytes as seen in other vertebrates. However, Xt-runx2 expression in enlarged chondrocytes was weak and dissimilar to common hypertrophic chondrocytes. These observations suggest that weak Xt-runx2 expression in maturing chondrocytes affects characteristic bone development in X. tropicalis long bones.