Transcription and immunolocalization of Runx2/Cbfa1/Pebp2αA in developing rodent and human craniofacial tissues:: Further evidence suggesting osteoclasts phagocytose osteocytes

Runx2/Cbfa1 is a transcription factor, essential for the osteogenic/chondrogenic and odontogenic lineage. Three isoforms of Cbfa1 have been identified, type I (Pebp2alphaA isoform), type II (til-1 isoform), and type III (Osf2 isoform). Here we examined the expression of the Runx2/Cbfa1 during intramembranous and enchondral bone formation in the craniofacial tissues of neonatal rodents (hamster, rat, mouse) and the human fetus. We used a monoclonal antibody raised against the Pebp2alphaA portion and thus potentially recognizing all three isoforms of Runx2/Cbaf1. We report Cbfa1 at the mRNA and protein level in periosteum, preosteoblasts, osteoblasts, young osteocytes, perichondrium, resting and hypertrophic chondrocytes. During active bone remodeling, almost one third of tartrate resistant acid phosphatase (TRAP) positive multinuclear cells identified as osteoclasts were also stained with anti-Pebp2alphaA antibodies. Osteoclasts, however, did not express mRNA transcripts of the Pebp2alphaA gene. Some of the immunopositive structures within these osteoclasts resembled (ingested) cells. TRAP-positive mononuclear cells not attached to bone surfaces did not stain with anti-Pebp2alphaA antibodies. We concluded that the tissue distribution of Runx2/Cbaf1/Pebp2alphaA in ossifying bones of the human fetus is similar to that in neonatal rodent tissues. Osteoclasts do not transcribe the Runx2/Cbfa1 gene but become immunostained by phagocytosing and digesting osteocytes/hypertrophic chondrocytes. The substantial number of osteoclasts involved in phagocytosis of Runx2/Cbfa1 immunopositive cells suggests that phagocytosis is a major way of removing osteocytes/hypertrophic chondrocytes during resorption of bone and cartilage. Finally, the data indicate that positive immunostaining of osteoclasts for typical osteogenic/chondrogenic markers has to be interpreted with caution due to the phagocytosing capacity of these cells. (C) 2003 Wiley-Liss, Inc.