Parathyroid hormone and other agents that stimulate bone resorption function, at least in part, by inducing osteoblasts to secrete cytokines that stimulate osteoclast differentiation and activity. We previously demonstrated that parathyroid hormone induces expression by osteoblasts of interleukin-6 and leukemia inhibitory factor without affecting the 16 other cytokines that were examined. We also showed that stimulation of osteoclast activity by parathyroid hormone is dependent on activation of the cAMP signal transduction pathway and secretion of interleukin-6 by osteoblasts. In the current study, we demonstrate that the rapid and transient stimulation of interleukin-6 and leukemia inhibitory factor is inhibited by actinomycin D and superinduced by protein synthesis inhibitors, the classical characteristics of an immediate-early gene response. Moreover, activation of cAMP signal transduction by parathyroid hormone and parathyroid hormone-related protein is necessary and sufficient to induce both interleukin-6 and leukemia inhibitory factor. In addition, cAMP analogues as well as vasoactive intestinal peptide and isoproterenol, two neuropeptides that stimulate bone resorption by activating cAMP signal transduction in osteoblasts, also induce interleukin-6 and leukemia inhibitory factor in these cells. Taken together with our previous results, this study suggests that interleukin-6 is crucial for stimulation of bone resorption not only by parathyroid hormone, but also by parathyroid hormonerelated protein, vasoactive intestinal peptide, and -adrenergic agonists, like isoproterenol.Excessive skeletal loss is caused by a disturbance in the normal balance between bone resorption by osteoclasts, and bone formation by osteoblasts. Knowledge of the mechanisms that regulate these processes is fundamental to the understanding of the pathogenesis of bone loss that occurs in conditions such as osteoporosis. In addition to forming bone, osteoblasts appear to mediate the effect of parathyroid hormone (PTH) 1 and other resorptive agents (1, 2). This concept is best supported by evidence that many agents, including PTH, stimulate bone resorption by osteoclasts that are cultured in the presence of osteoblasts but not by osteoclasts that are cultured alone. In most cases, resorption is also stimulated if the osteoblasts are replaced by conditioned media from osteoblasts activated by the resorptive agents. It is therefore thought that the primary effect of the resorptive agents is to stimulate osteoblasts to produce soluble cytokines that activate the osteoclasts.We previously examined the effect of PTH on expression by osteoblasts of mRNAs encoding 18 cytokines, and found that of these only interleukin-6 (IL-6) and leukemia inhibitory factor (LIF) were stimulated by the hormone (3, 4). Maximal stimulation was approximately 50-fold for IL-6 and approximately 10-fold for LIF. PTH also stimulates secretion by osteoblasts of both IL-6 (3-9) and LIF (10) proteins. Lack of stimulation of IL-6 and LIF by PTH in particular cell prepara...