Stimulation of alkaline phosphatase activity in cultured neonatal mouse calvarial bone cells by parathyroid hormone

Abstract

The effect of parathyroid hormone (PTH) on alkaline phosphatase activity was examined in confluent, serum-free primary cultures of neonatal mouse calvarial cells. It was found that synthetic bPTH-(1-34) caused an increase in the specific activity of skeletal alkaline phosphatase isoenzyme by 18 hours. Between 10 and 500 ng/ml, the magnitude of the change was directly related to peptide concentration. The change occurred in the absence of any effect on cell number, total cell protein, or DNA and was not the result of an effect on either proliferation or survival of a specific cell population. Results of histochemical studies indicate that bPTH-(1-34) caused an increase in the proportion of cells containing enzyme activity. The response was duplicated by intact bPTH-(1-84) and DBcAMP, but not by oxidized bPTH-(1-34) or insulin and did not require prostaglandin synthesis or hydroxylation of 25-hydroxyvitamin D3. These results demonstrate that bPTH has a direct effect on osteoblast maturation in vitro, that the effect is specific for PTH, and suggest that it is mediated by cAMP.