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The number of parathyroid hormone (PTH) receptors in rat calvaria (RC) cells increased with culture duration, reached a maximum on day 13 when the cells had not yet differentiated into osteoblasts, and then gradually decreased by day 21. However, short exposure (48h) of RC cells to PTH-(1-34) before day 12, but not after day 15, did not decrease alkaline phosphatase (ALP) activity of the cells. Bone-like nodule (BN) formation was also suppressed only when PTH-(1-34) was present during the late culture period (days 14-21). Pretreating the cells with staurosporine, a protein kinase C inhibitor, on day 7 resulted in augmentation of PTH-(1-34)-induced cyclic adenosine monophosphate (cAMP) accumulation and inhibition of ALP activity. PTH-(7-34), which only activates PKC, however, had no effect on RC cell differentiation under any conditions used.[PUBLICATION ABSTRACT]