DETECTION OF TRANSCRIPTS AND BINDING-SITES FOR COLONY-STIMULATING FACTOR-I DURING BONE-DEVELOPMENT

Colony-stimulating factor-1 (CSF-1), originally characterized as the growth factor for the cells of the mononuclear phagocytic system, has been shown to be essential for osteoclast formation. The aim of the present study was twofold: (i) to investigate the expression of transcripts encoding CSF-1; and (ii) to detect binding sites for CSF-1 during bone development. As a model, metatarsal rudiments from embryonic mice of different ages were used, an in vivo system allowing one to follow osteoclast formation. In 16-day-old embryos, proliferating osteoclast precursors are located on the outer surface of the rudiments. They differentiate subsequently to post-mitotic precursors. At 18 days, the precursors fuse and the mature osteoclasts invade the mineralized cartilage of the rudiments to excavate the future bone marrow cavity. Within this study, in situ hybridization on sections of whole paws from 17-day-old embryos revealed CSF-1 transcripts to be present in cells lining the outside of the midregion of the metatarsals. One day later, cells containing CSF-1 mRNA were found within the mineralized cartilage. The levels of transcripts encoding CSF-1 were further increased in the bone rudiments of newborn animals. Binding sites for CSF-1 on cells in close proximity of the metatarsals were detected at embryonic age 17 days, but not before. At this stage, cells binding CSF-1 were located on the periosteum of the midregion of the metatarsal rudiment. At 18 days, cells expressing high levels of CSF-1 binding sites had invaded the mineralized cartilage. Furthermore, the growth factor was shown to be bound by isolated osteoclasts. These binding studies demonstrated that CSF-1 binding sites are present on osteoclast precursors and on the mature osteoclasts. In conclusion, the presented results suggest that CSF-1 might act directly on the cells of the osteoclastic lineage and that it is locally synthesized during the terminal differentiation steps of these cells.