Cartilage matrix protein (CMP) is a non-collagenous component of cartilage with a yet unknown function. In this study we used in situ hybridization to investigate the temporal and spatial distribution of CMP transcripts during human embryonic and early fetal development, and compared it to the pattern of expression observed for collagen types I, II, X, and decorin. The distribution of CMP and collagen type II transcripts followed a similar pattern in the embryonic bone anlage, the fetal growth plate, and the developing vertebral column. Expression was highest in the upper hypertrophic and lower proliferative zone, whereas calcified cartilage was negative throughout the different stages of bone development. Chondrocytes of calcified cartilage, however, were not quiescent but expressed collagen type X. The onset of collagen type X expression was linked to hypertrophy and occurred before calcification became apparent. In contrast, decorin and collagen type I were highly expressed in bone and perichondrium but not in growth plate cartilage. During the development of the synovial joints a different pattern of expression emerged. After formation of the joint cavity, there was a halt in expression of CMP but not of collagen type II in chondrocytes close to the articular surface. A band of CMP negative chondrocytes covering the joint surface was observed in all joints investigated. Decorin mRNA was demonstrated in the reserve zone adjacent to the joints, but not in articular cartilage. Extraskeletal expression of CMP was observed in the embryonic retina. The results demonstrate the differential expression of CMP during human skeletal development and chondrocyte differentiation. The distribution of CMP transcripts is unique and distinct from other known matrix genes.