Carbonic anhydrase II gene transcript in cultured osteoclasts from neonatal rats: effect of calcitonin

Carbonic anhydrase II (CA II), an enzyme catalyzing the interconversion of CO2 and water to HCO3- and protons, has a key role in osteoclastic bone resorption, but little is known of the regulation of CA II gene expression by calcitonin. Analysis of mRNA in osteoclasts has been difficult because of the problems of obtaining sufficient number of purified osteoclasts from bone. In this study, however, we have investigated the regulation of CA II mRNA in rat osteoclasts and their putative mononuclear precursors by using in situ hybridization. We have found that the CA II gene is expressed at high levels in osteoclasts and what are probably their maturing mononuclear precursors. Measurement of CA II mRNA in cultured osteoclasts and their putative mononuclear precursor cells by cytophotometry provided evidence that calcitonin, a direct inhibitor of mammalian osteoclast activity, reduces the levels of CA II mRNA in a dose dependent manner; maximum reduction was observed at a concentration of 100pM of calcitonin. In addition, calcitonin reduced the number of CA II mRNA-positive mononuclear precursor cells. The results also suggest that expression of the CA II gene is a feature of cells committed to the osteoclast lineage.