Tonic inhibitory role for cAMP in α1a-adrenergic receptor coupling to extracellular signal-regulated kinases 1/2

Xiuxiang Jiao, Pedro J. Gonzalez-Cabrera, Lei Xiao, Michael E. Bradley, Peter W. Abel, William B. Jeffries

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Abstract

α1a-Adrenergic receptors (ARs) couple to phosphoinositide hydrolysis, adenylyl cyclase, and mitogen-activated protein kinase (MAPK) pathways. However, the interaction among these signaling pathways in activating extracellular signal-regulated kinase 1/2 (ERK1/2) is not well understood. We investigated the coupling of α1a-ARs to ERK1/2 in Chinese hamster ovary (CHO)-K1 cells stably transfected with mouse α1a-ARs, as well as the interaction between ERK1/2 and norepinephrine-induced cAMP accumulation. α1a-AR activation by norepinephrine increased the cytosolic Ca2+ concentration and phosphorylated ERK1/2 in a time- and concentration-dependent manner. ERK1/2 phosphorylation was blocked by the MAPK kinase 1/2 inhibitor 2′-amino-3′-methoxyflavone (PD 98059) and the α1a-AR antagonist prazosin. A transient elevation in intracellular Ca2+ was required for the phosphorylation of ERK1/2; however, activation of protein kinase C did not seem to be required for ERK1/2 phosphorylation. Norepinephrine also stimulated cAMP accumulation in transfected CHO-K1 cells in a concentration-dependent manner via α1a-ARs, which was blocked by the Ca2+ chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid. Norepinephrine-induced ERK1/2 phosphorylation was inhibited by the adenylyl cyclase activator forskolin and was enhanced by the adenylyl cyclase inhibitor 9-(tetrahydro-2-furanyl)-9H-purine-6-amine (SQ 22536) and the protein kinase A inhibitor 4-cyano-3-methylisoquinoline. In conclusion, in transfected CHO-K1 cells, α1a-AR activation activates both phospholipase C and adenylyl cyclase-mediated signaling pathways. α1a-AR-mediated ERK1/2 phosphorylation was dependent on a rise in intracellular Ca2+, and this pathway was reciprocally regulated by the concomitant activation of adenylyl cyclase, which inhibits ERK1/2 phosphorylation. Thus, α1a-AR stimulation of cAMP production may play an important role in regulating ERK1/2 phosphorylation in cell lines and native tissues.

Original languageEnglish
Pages (from-to)247-256
Number of pages10
JournalJournal of Pharmacology and Experimental Therapeutics
Volume303
Issue number1
DOIs
StatePublished - Oct 2002

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Mitogen-Activated Protein Kinase 3
Mitogen-Activated Protein Kinase 1
Adrenergic Receptors
Phosphorylation
Adenylyl Cyclases
Norepinephrine
Cricetulus
Ovary
MAP Kinase Kinase 2
MAP Kinase Kinase 1
Ethane
Adrenergic Antagonists
Prazosin
Type C Phospholipases
Colforsin
Protein Kinase Inhibitors
Chelating Agents
Phosphatidylinositols
Cyclic AMP-Dependent Protein Kinases
Mitogen-Activated Protein Kinases

All Science Journal Classification (ASJC) codes

  • Pharmacology

Cite this

Tonic inhibitory role for cAMP in α1a-adrenergic receptor coupling to extracellular signal-regulated kinases 1/2. / Jiao, Xiuxiang; Gonzalez-Cabrera, Pedro J.; Xiao, Lei; Bradley, Michael E.; Abel, Peter W.; Jeffries, William B.

In: Journal of Pharmacology and Experimental Therapeutics, Vol. 303, No. 1, 10.2002, p. 247-256.

Research output: Contribution to journalArticle

Jiao, Xiuxiang ; Gonzalez-Cabrera, Pedro J. ; Xiao, Lei ; Bradley, Michael E. ; Abel, Peter W. ; Jeffries, William B. / Tonic inhibitory role for cAMP in α1a-adrenergic receptor coupling to extracellular signal-regulated kinases 1/2. In: Journal of Pharmacology and Experimental Therapeutics. 2002 ; Vol. 303, No. 1. pp. 247-256.
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AU - Jeffries, William B.

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N2 - α1a-Adrenergic receptors (ARs) couple to phosphoinositide hydrolysis, adenylyl cyclase, and mitogen-activated protein kinase (MAPK) pathways. However, the interaction among these signaling pathways in activating extracellular signal-regulated kinase 1/2 (ERK1/2) is not well understood. We investigated the coupling of α1a-ARs to ERK1/2 in Chinese hamster ovary (CHO)-K1 cells stably transfected with mouse α1a-ARs, as well as the interaction between ERK1/2 and norepinephrine-induced cAMP accumulation. α1a-AR activation by norepinephrine increased the cytosolic Ca2+ concentration and phosphorylated ERK1/2 in a time- and concentration-dependent manner. ERK1/2 phosphorylation was blocked by the MAPK kinase 1/2 inhibitor 2′-amino-3′-methoxyflavone (PD 98059) and the α1a-AR antagonist prazosin. A transient elevation in intracellular Ca2+ was required for the phosphorylation of ERK1/2; however, activation of protein kinase C did not seem to be required for ERK1/2 phosphorylation. Norepinephrine also stimulated cAMP accumulation in transfected CHO-K1 cells in a concentration-dependent manner via α1a-ARs, which was blocked by the Ca2+ chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid. Norepinephrine-induced ERK1/2 phosphorylation was inhibited by the adenylyl cyclase activator forskolin and was enhanced by the adenylyl cyclase inhibitor 9-(tetrahydro-2-furanyl)-9H-purine-6-amine (SQ 22536) and the protein kinase A inhibitor 4-cyano-3-methylisoquinoline. In conclusion, in transfected CHO-K1 cells, α1a-AR activation activates both phospholipase C and adenylyl cyclase-mediated signaling pathways. α1a-AR-mediated ERK1/2 phosphorylation was dependent on a rise in intracellular Ca2+, and this pathway was reciprocally regulated by the concomitant activation of adenylyl cyclase, which inhibits ERK1/2 phosphorylation. Thus, α1a-AR stimulation of cAMP production may play an important role in regulating ERK1/2 phosphorylation in cell lines and native tissues.

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