Parathyroid hormone-induced ornithine decarboxylase activity in fetal rat osteoblasts.
Induction of ornithine decarboxylase (ODC, E.C. 4.1.1.17) activity by parathyroid hormone (PTH) in cultured fetal rat osteoblasts was studied. PTH induced ODC activity and stimulated cAMP production in a dose-dependent manner, the ED50 for cAMP being five times as high as that for ODC. Induction of ODC activity by PTH was partly inhibited by actinomycin D and cycloheximide, with 40 and 55% inhibition, respectively. PTH increased the intracellular ionized calcium concentration ([Ca2+]i), which was absent in a Ca2+-free medium. Blocking calcium influx, lowering the extracellular calcium concentration, and adding trifluoperazine inhibited both induction of ODC activity and stimulation of cAMP production by PTH. A23187 (100 nM and 1 microM), combined with a low dose of PTH (4 nM), resulted in a synergistic induction of ODC activity and an inhibition of cAMP production. A23187 inhibited induction of ODC activity as well as stimulation of cAMP production by the dose of PTH (20 nM) maximally effective in inducing ODC activity. Forskolin together with this maximal dose of PTH resulted in an additive effect on ODC activity and a synergistic stimulation of cAMP production. The current results show similarities and differences with respect to results obtained with osteoblasts from other species and osteoblast cell lines. The present data indicate that (1) PTH stimulates ODC activity and this is partly due to new enzyme synthesis; (2) calcium is involved in induction of ODC activity and stimulation of cAMP production by PTH; furthermore, it is suggestive that calmodulin and/or protein kinase C are involved; and (3) stimulation of cAMP production by PTH depends on an optimal intracellular calcium concentration range.
Van Leeuwen
J P
JP
Laboratory for Cell Biology and Histology, University of Leiden, The Netherlands.
Bos
M P
MP
Herrmann-Erlee
M P
MP
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002478:Cells, Cultured; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism; D003513:Cycloheximide / Q000494:pharmacology; D003609:Dactinomycin / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004790:Enzyme Induction; D005865:Gestational Age; D007700:Kinetics; D009955:Ornithine Decarboxylase / Q000096:biosynthesis* / Q000378:metabolism; D065108:Ornithine Decarboxylase Inhibitors; D010006:Osteoblasts / Q000201:enzymology*; D010281:Parathyroid Hormone / Q000494:pharmacology*; D051381:Rats; D015290:Second Messenger Systems; D014268:Trifluoperazine / Q000494:pharmacology; D014700:Verapamil / Q000494:pharmacology
D002121:Calcium Channel Blockers; D065108:Ornithine Decarboxylase Inhibitors; D010281:Parathyroid Hormone; D003609:Dactinomycin; D005576:Colforsin; D014268:Trifluoperazine; D003513:Cycloheximide; D014700:Verapamil; D000242:Cyclic AMP; D009955:Ornithine Decarboxylase; D002118:Calcium