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Am J Physiol Renal Physiol (January 29, 2002). doi:10.1152/ajprenal.00318.2001
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Articles in PresS, published online ahead of print January 28, 2002
Am J Physiol Renal Physiol, 10.1152/ajprenal.00318.2001
Submitted on October 23, 2001
Accepted on December 8, 2001

Role of cAMP-PKA-PLC signaling cascade on dopamine-induced PKC-mediated inhibition of renal Na+-K+-ATPase activity

Pedro Gomes1 and Patricio Soares-da-Silva1*

1 Inst. Pharmacol. Ther., Fac. Medicine, Porto, Portugal

The molecular events set into motion from stimulation of D1-like receptors downstream to Na+-K+-ATPase were investigated while measuring apical-to-basal ouabain-sensitive amphotericin B-induced increases in short circuit current in opossum kidney (OK) cells. The D1-like receptor agonist SKF 38393 decreased Na+-K+-ATPase activity with an IC50 value of 130 nM. This effect was prevented either by the D1-like receptor antagonist SKF 83566, overnight treatment with cholera toxin, the PKA antagonist H-89, or the PKC antagonist chelerythrine, but not by the MAPK inhibitor PD 098059 or the phosphatidylinositol 3-kinase inhibitors wortmannin and LY 294002. Both dibutyril cAMP (db-cAMP) and phorbol-12,13-dibutyrate (PDBu) effectively reduced Na+-K+-ATPase activity. Down regulation of PKA abolished the inhibitory effects of SKF 38393 and db-cAMP upon Na+-K+-ATPase activity, but not that of PDBu. Down regulation of PKC abolished the inhibitory effects of PDBu, SKF 38393 and db-cAMP. The phospholipase C (PLC) inhibitor, U-73,122, prevented the inhibitory effects of both SKF 38393 and db-cAMP. On the other hand, db-cAMP was found to increase PLC activity. Though OK cells were found to express both Gs{alpha} and Gq/11{alpha} proteins, D1-like receptors appear to be coupled to Gs{alpha} proteins only, as evidenced in functional studies using cells treated overnight with specific antibodies raised against Gs{alpha} and Gq/11{alpha} proteins. It is concluded that PLC and Na+-K+-ATPase are effector proteins for PKA and PKC, respectively, following stimulation of D1-like receptors coupled to Gsalpha proteins, in a sequence of events that begins with the activation of the adenylyl cyclase-PKA system followed by activation of the PLC-PKC system.




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