alpha(2)-Adrenergic receptors activate MAPK and Akt through a pathway involving arachidonic acid metabolism by cytochrome P450-dependent epoxygenase, matrix metalloproteinase activation and subtype-specific transactivation of EGFR.
Previous study carried out on PC12 cells expressing each alpha(2)-adrenergic receptor subtype individually (PC12/alpha(2A), /alpha(2B) or /alpha(2C)) have shown that epinephrine causes activation of PI3K and phosphorylation of Erk 1/2. The signal transduction mechanisms whereby each alpha(2)-AR subtype triggers these actions were investigated in the present study. In all three clones, epinephrine-induced phosphorylation of MAPK or Akt was abolished by prior treatment with ketoconazole, but not with indomethacin or nordihydroguaiaretic acid. On the other hand, treatment of the clones with epinephrine caused a rapid increase of AA release, which was fully abolished by the PLC inhibitor U73122, but was unaffected by the PLA(2) inhibitor quinacrine. The effects of epinephrine on MAPK and Akt were mimicked by cell exposure to exogenous AA. Furthermore, whereas U73122 abolished the effects of epinephrine, quinacrine only prevented the effects of epinephrine, suggesting that AA release through PLC and its metabolites are responsible for MAPK and Akt activation by alpha(2)-ARs. Treatment with 1,10-phenanthroline, CRM197, or tyrphostin AG1478 suppressed MAPK and Akt phosphorylation by epinephrine or AA, in a subtype-specific manner. Furthermore, conditioned culture medium from epinephrine-treated PC12/alpha(2) induced MAPK and Akt phosphorylation in wild-type PC12. Inhibition of NGFR tyrosine phosphorylation had no effect but the src inhibitor PP1 abolished MAPK and Akt phosphorylation in all three clones. Our results provide evidence for a putative pathway by which alpha(2)-ARs activate MAPK and Akt in PC12 cells, involving stimulation of PLC, AA release, AA metabolism by cytochrome P450-dependent epoxygenase, stimulation of matrix metalloproteinases and subtype-specific transactivation of EGFR through src activation and heparin-binding EGF-like growth factor release.
Karkoulias
Georgios
G
Department of Pharmacology, School of Medicine, University of Patras, 26504 Rio Patras, Greece.
Mastrogianni
Orthodoxia
O
Lymperopoulos
Anastasios
A
Paris
Herve
H
Flordellis
Christodoulos
C
D000316:Adrenergic alpha-Agonists / Q000378:metabolism; D000818:Animals; D000935:Antifungal Agents / Q000378:metabolism; D016718:Arachidonic Acid / Q000378:metabolism*; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000378:metabolism; D004837:Epinephrine / Q000378:metabolism; D066246:ErbB Receptors / Q000378:metabolism*; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism*; D006801:Humans; D007654:Ketoconazole / Q000378:metabolism; D020935:MAP Kinase Signaling System / Q000502:physiology*; D020782:Matrix Metalloproteinases / Q000378:metabolism*; D010088:Oxidoreductases / Q000378:metabolism*; D016716:PC12 Cells; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics / Q000378:metabolism*; D011796:Quinacrine / Q000378:metabolism; D051381:Rats; D020917:Receptor, trkA / Q000378:metabolism; D018341:Receptors, Adrenergic, alpha-2 / Q000235:genetics / Q000378:metabolism*; D015533:Transcriptional Activation; D019061:src-Family Kinases / Q000378:metabolism
D000316:Adrenergic alpha-Agonists; D000935:Antifungal Agents; D004791:Enzyme Inhibitors; D020033:Protein Isoforms; D018341:Receptors, Adrenergic, alpha-2; D016718:Arachidonic Acid; D003577:Cytochrome P-450 Enzyme System; D010088:Oxidoreductases; D019869:Phosphatidylinositol 3-Kinases; D066246:ErbB Receptors; D020917:Receptor, trkA; D019061:src-Family Kinases; D051057:Proto-Oncogene Proteins c-akt; D048049:Extracellular Signal-Regulated MAP Kinases; D020782:Matrix Metalloproteinases; D011796:Quinacrine; D007654:Ketoconazole; D004837:Epinephrine