Parathyroid hormone inhibits collagen synthesis and the activity of rat col1a1 transgenes mainly by a cAMP-mediated pathway in mouse calvariae.
We examined the effect of parathyroid hormone and various signaling molecules on collagen synthesis and chloramphenicol acetyltransferase activity in cultured transgenic mouse calvariae carrying fusion genes of the rat Col1a1 promoter and the chloramphenicol acetyltransferase reporter. After 48 h of culture, parathyroid hormone, forskolin, dibutyryl cAMP, 8-bromo cAMP, and phorbol myristate acetate inhibited transgene activity, while the calcium ionophore ionomycin had no effect. Pretreatment of calvariae with the phosphodiesterase inhibitor isobutylmethylxanthine potentiated the inhibitory effect of 1 nM parathyroid hormone on transgene activity and collagen synthesis. Parathyroid hormone further inhibited transgene activity and collagen synthesis in the presence of phorbol myristate acetate. Parathyroid hormone inhibition of transgene activity and collagen synthesis was not affected by indomethacin or interleukin-6. After 48 h of culture, parathyroid hormone inhibited chloramphenicol acetyltransferase activity by 50-85% in cultured calvariae carrying transgenes having progressive 5' upstream deletions of promoter DNA down to -1683 bp. These data show that the inhibitory effect of parathyroid hormone on Col1a1 expression in mouse calvariae is mediated mainly by the cAMP signaling pathway. Prostaglandins and IL-6 are not local mediators of the parathyroid hormone response in this model. Finally, regions of the Col1a1 promoter downstream of -1683 bp are sufficient for parathyroid hormone inhibition of the Col1a1 promoter.
Bogdanovic
Z
Z
Department of Medicine, School of Medicine, University of Connecticut Health Center, Farmington, Connecticut 06030, USA.
Huang
Y F
YF
Dodig
M
M
Clark
S H
SH
Lichtler
A C
AC
Kream
B E
BE
D000818:Animals; D003094:Collagen / Q000096:biosynthesis* / Q000235:genetics*; D000242:Cyclic AMP / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D010281:Parathyroid Hormone / Q000494:pharmacology*; D011347:Procollagen / Q000235:genetics; D051381:Rats; D015398:Signal Transduction / Q000187:drug effects; D012886:Skull / Q000378:metabolism*
D010281:Parathyroid Hormone; D011347:Procollagen; D003094:Collagen; D000242:Cyclic AMP