mu-Opioids activate tyrosine kinase focal adhesion kinase and regulate cortical cytoskeleton proteins cortactin and vinculin in chick embryonic neurons.
We have investigated the signal transduction pathway of the G-protein mu-opioid receptor upstream of phospholipase D (PLD) and protein kinase C-epsilon (PKC-epsilon) activation in postmitotic E6CH chick embryo cortical neurons. The mu-opioid receptor and PLD-PKC-epsilon functional coupling depends on upstream tyrosine kinase activation. We now report that the mu-opioid agonists specifically stimulated tyrosine phosphorylation and activation of the focal adhesion kinase (FAK) in a time-dependent manner. We also demonstrate that met-enkephalin, a mu-opioid agonist in E6CH cultures, significantly increases tyrosine phosphorylation of another Src kinase substrate, the cytoskeletal protein cortactin. Tyrosine phosphorylation of cortactin led to drastic changes in subcellular localization, an estimated 2-fold enrichment in the cytosol. Similarly, opioids stimulated a sustained tyrosine phosphorylation of vinculin, a protein enriched in focal adhesion sites. These data provide novel evidence that opioid receptor intracellular signaling engages the specific activation of tyrosine kinase FAK and regulates the neuronal cytoskeleton during central nervous system morphogenesis.
Mangoura
D
D
Department of Pediatrics, University of Chicago, IL 60637, USA. dm36@midway.uchicago.edu
D000818:Animals; D015815:Cell Adhesion Molecules / Q000096:biosynthesis / Q000378:metabolism*; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology / Q000196:embryology; D002642:Chick Embryo; D051356:Cortactin; D003598:Cytoskeletal Proteins / Q000096:biosynthesis / Q000378:metabolism*; D004744:Enkephalin, Methionine / Q000494:pharmacology*; D004789:Enzyme Activation; D051416:Focal Adhesion Protein-Tyrosine Kinases; D007700:Kinetics; D008840:Microfilament Proteins / Q000096:biosynthesis / Q000378:metabolism*; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000096:biosynthesis / Q000378:metabolism*; D017450:Receptors, Opioid, mu / Q000187:drug effects / Q000502:physiology*; D016596:Vinculin / Q000096:biosynthesis / Q000378:metabolism*
D015815:Cell Adhesion Molecules; D051356:Cortactin; D003598:Cytoskeletal Proteins; D008840:Microfilament Proteins; D017450:Receptors, Opioid, mu; D016596:Vinculin; D019000:Phosphotyrosine; D004744:Enkephalin, Methionine; D011505:Protein-Tyrosine Kinases; D051416:Focal Adhesion Protein-Tyrosine Kinases