Triiodothyronine stimulates the release of membrane-bound alkaline phosphatase in osteoblastic cells

Thyroid hormone deficient osteoblastic cells in cell culture released a significantly higher amount of alkaline phosphate (ALP) activity following T3 replacement. T3 increased the release of total and membrane-bound ALP activity in these cells significantly more than T4 or inactive thyroid hormone metabolite, DIT. The effect of T3 on the membrane-bound ALP fraction was dose and time dependent; higher concentrations of T3 and longer incubation time with T3 proportionally increased the enzyme activity. T3 had no effect on the release of soluble fraction of ALP. Our results indicate that in "hypothyroid" osteoblastic cells the total release of ALP is decreased and that the secreted fraction of ALP is predominantly in soluble form, whereas the addition of T3 stimulates ALP release and mainly increases the membrane-bound fraction. T3 also increased formation of actin cytoskeleton in hypothyroid osteoblastic cells. Cytochalasin treatment, through its inhibition of actin polymerization, produced a significant decrease of membrane-bound ALP release induced by T3. These data suggest that the regulatory role of T3 in skeletal development can partly be due to its stimulatory effect on the release of membrane-bound ALP by osteoblastic cells which is thought to be an important factor in the initiation of biological calcification.