Regulatory effect of calcium-regulating hormones on the synthesis and/or secretion of bone .GAMMA.-carboxyglutamic acid-containing protein in chick embryonic calvaria in vitro.

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Regulatory effects of 1, 25-dihydroxyvitamin D3 (1, 25-(OH)2-D3), 24, 25-dihydroxyvitamin D3 (24, 25-(OH)2-D3), parathyroid' hormone (PTH) and calcitonin (CT) on BGP synthesis and/or secretion were studied using chick embryonic calvaria in vitro. BGP contents in calvaria and culture medium were determined by radioimmunoassay using antiserum to purified chick BGP. After 72 h culture, 1, 25-(OH)2-D3 (10-10-10-7M) increased the BGP content in culture medium signi-ficantly, and the effect was maximum at 10-8M, while the BGP content in calvaria was not changed by 1, 25-(OH)2-D3.24, 25-(OH)2-D3 increased the BGP content both in medium at 10-6M-10-5M after 24h culture and in calvaria at 5×10-7-10-5M after 48h culture. PTH increased BGP contents both in calvaria after 72 h culture and medium at 1U/ml after 24h culture, while it decreased them at 5-10 U/ml after 24 h culture. CT (0.5-10U/ml) had no effect on BGP contents in calvaria and medium. 1, 25-(OH)2-D3-induced increase of BGP in culture medium was observed from 24 to 72 h of culture and then reached a plateau at 120h of culture. 1, 25-(OH)2-D3-induced increase in the BGP level in medium after 72h culture was 3-4 times the control value. On the other hand, the BGP content in calvaria was not affected by 1, 25-(OH)2-D3 until 72h of culture and then significantly increased at 120h. The addition of 1, 25-(OH)2-D3 (10-8M), 24, 25-(OH)2-D3 (10-6M), and 1U PTH (and 1 U. CT added to those hormones) into culture medium of chick embryonic calvaria signi-ficantly increased BGP contents in calvaria and medium at 72h after culture. The effect of three hormones was not synergistic, but had a tendency to be greater than that of a single addition of 1, 25-(OH)2-D3, 24, 25-(OH)2-D3 or PTH. Further addition of 1 U CT into culture medium did not affect the combined effects of 1, 25-(OH)2-D3, 24, 25-(OH)2-D3 and PTH except for the calcium content in calvaria. The addition of individual calcium-regulating hormones into culture medium of chick embryonic calvaria did not affect the calcium and phosphorus contents in calvaria, but the calcium content in calvaria was significantly increased by the addition of 1, 25-(OH)2-D3, 24, 25-(OH)2-D3 and PTH to the culture medium. Actinomycin D completely eliminated the 1, 25-(OH)2-D3-, 24, 25-(OH)2-D3- or PTH-induced increase in BGP, but had no appreciable effect on basal BGP production. These results support a model of transcriptional control for the action of those hormones on BGP synthesis. These results suggest that the effects of calcium-regulating hormones on bone will be exerted via their actions on BGP synthesis and/or secretion, respectively.
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Regulatory effect of calcium-regulating hormones on the synthesis and/or secretion of bone .GAMMA.-carboxyglutamic acid-containing protein in chick embryonic calvaria in vitro.

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