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Am J Physiol Cell Physiol 283: C1432-C1440, 2002. First published July 3, 2002; doi:10.1152/ajpcell.00221.2002
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Vol. 283, Issue 5, C1432-C1440, November 2002

Termination of immediate-early gene expression after stimulation by parathyroid hormone or isoproterenol

Xin Chen1, Jia-Chun Dai1, and Edward M. Greenfield1,2,3

Departments of 1 Orthopaedics, 2 Pathology, and 3 Physiology and Biophysics, Case Western Reserve University and University Hospitals of Cleveland, Cleveland, Ohio 44106-5000

cAMP/PKA signaling transiently stimulates mRNA expression of immediate-early genes, including IL-6 and c-fos. We confirmed that these mRNAs are transiently stimulated by parathyroid hormone (PTH) in ROS 17/2.8 osteoblastic cells. Consistent with the role for cAMP/PKA signaling in this response, PTH induces transient cAMP elevation, PKA activation, and cAMP-responsive element-binding protein (CREB) phosphorylation. Our goal was to determine whether termination of immediate-early gene expression is due to receptor desensitization or cAMP degradation. The approaches used were 1) inhibition of PTH receptor desensitization with G protein-coupled receptor kinase 2 (GRK2) antisense oligonucleotides or antisense plasmids, 2) sustained activation of adenyl cyclase with forskolin, and 3) inhibition of cAMP degradation with 3-isobutyl-1-methylxanthine. These experiments show that mechanisms downstream of receptor desensitization and cAMP degradation are primarily responsible for termination of PKA activity, CREB phosphorylation, and immediate-early gene expression. Similar conclusions were also obtained in response to PTH in a second osteoblastic cell line (MC3T3-E1) and in response to isoproterenol in NIH3T3 fibroblasts. This conclusion may therefore reflect a general mechanism for termination of immediate-early gene expression after induction by cAMP/PKA.

receptor desensitization; cAMP degradation; IL-6; c-fos; osteoblast


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