Identification of regulatory molecules for osteogenic activity of BMP.

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Bone morphogenetic proteins (BMPs) regulate many aspects of skeletal development, including osteoblast and chondrocyte differentiation, cartilage and bone formation, and cranial and limb development. Among them, BMP-2, one of the most potent osteogenic signaling molecules, stimulates osteoblast differentiation, but inhibits myogenic differentiation in C2C12 cells. To evaluate important genes for BMP-2-induced osteoblast differentiation, we performed cDNA microarray analysis between BMP-2 treated and untreated C2C12 cells, and identified Alx3 (aristaless like homeobox 3) as a gene that was clearly induced during osteoblast differentiation. Alx3 is a homeobox gene related to the Drosophila aristaless gene and a group of vertebrate genes that includes Prx1/2, Cart1 and Alx4. Alx3 is expressed in frontonasal head mesenchyme and the first and second pharyngeal arches. Alx3 has been linked to developmental functions in craniofacial structures and limb development. Little is known about its direct relationship with bone formation. In the present study, we also focused on the mechanisms of Alx3 gene expression and function during osteoblast differentiation induced by BMP-2.