Parathyroid hormone (PTH) and PTH-related protein interact with a G protein-coupled receptor linked to the activation of adenylyl cyclase and phospholipase C signaling pathways. Regulation by PTH of the expression of three distinct, stably transfected luciferase reporter genes responsive to cAMP (CRE-luc), serum (SRE-luc) and phorbol ester (TRE-luc) has been studied in rat osteoblast-like UMR-106 cells. Maximal 43-fold induction of CRE-luc expression occurred in response to 100 nM rat (r)PTH(1-34) (EC50=0.44 nM), but SRE-luc and TRE-luc remained unaffected. Maximal 2.8- and 3.4-fold inductions of SRE-luc by 10 ng/ml EGF and 100 nM phorbol ester (PMA) were suppressed with 100 nM rPTH(1-34) (IC50=0.04 and 0.15 nM, respectively). Similarly, 7.3-fold induction of TRE-luc by 100 nM PMA was inhibited to 50% with 100 nM rPTH(1-34) (IC50=0.5 nM). Activation of mitogen-activated protein kinase by EGF and PMA was also suppressed by rPTH(1-34). 1 mM 8-Br-cAMP and 0.1 mM forskolin mimicked all the effects of rPTH(1-34). In conclusion, the regulation of target genes by PTH in osteoblast-like UMR-106 cells is mediated by the activation of the cAMP/protein kinase A signaling pathway.