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Am J Physiol Cell Physiol (July 3, 2002). doi:10.1152/ajpcell.00221.2002
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Articles in PresS, published online ahead of print July 3, 2002
Am J Physiol Cell Physiol, 10.1152/ajpcell.00221.2002
Submitted on May 16, 2002
Accepted on July 1, 2002

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

Xin Chen1, Jia-Chun Dai1, and Edward M Greenfield2*

1 Orthopaedics, Case Western Reserve University, Cleveland, OH, USA
2 Orthopaedics, Case Western Reserve University, Cleveland, OH, USA; Pathology, Case Western Reserve University, Cleveland, OH, USA; Physiology and Biophysics, Case Western Reserve University, Cleveland, OH, USA

* To whom correspondence should be addressed. E-mail: emg3{at}po.cwru.edu.

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 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 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 GRK2 antisense oligonucleotides or antisense plasmids, (2) sustained activation of adenyl cyclase with forskolin, and (3) inhibition of cAMP degradation with IBMX. 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 following induction by cAMP/PKA.




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