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Articles in PresS, published online ahead of print May 29, 2002
Am J Physiol Cell Physiol, 10.1152/ajpcell.00096.2002
Submitted on March 4, 2002
Accepted on May 23, 2002
1 Physiology, The University of Tennessee Health Science Center, Memphis, TN, USA
* To whom correspondence should be addressed. E-mail: thomason{at}physio1.utmem.edu.
Skeletal muscle Na+-K+-2Cl- cotransporter (NKCC) activity provides a potential mechanism for regulated potassium uptake. {{beta}}-Adrenergic receptor (ß-AR) activation stimulates skeletal muscle NKCC activity in an MAPK pathway-dependent manner. We examined potential G-protein coupled pathways for ß-AR-stimulated NKCC activity. Inhibition of Gs-coupled PKA blocked isoproterenol-stimulated NKCC activity in both the slow-twitch soleus muscle and the fast-twitch plantaris muscle. However, PKA-activating agents cholera toxin, forskolin, and 8-Br-cAMP were not sufficient to activate NKCC in the plantaris, and partially stimulated NKCC activity in the soleus. Isoproterenol-stimulated NKCC activity in the soleus was abolished by pretreatment with pertussis toxin (PTX), indicating a Gi-coupled mechanism. PTX did not affect the 8-Br-cAMP-stimulated NKCC activity. PTX treatment also precluded the isoproterenol-mediated ERK1,2 MAPK phosphorylation in the soleus, consistent with NKCC's MAPK dependency. Inhibition of isoproterenol-stimulated ERK activity by PTX treatment was associated with an increase in Akt activation and phosphorylation of Raf-1 on the inhibitory residue Ser259. These results demonstrate a novel, muscle phenotype-dependent mechanism for ß-AR-mediated NKCC activation that involves both Gs- and Gi-protein coupled mechanisms.
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