title: Coupling of ionic events to protein kinase signaling cascades upon activation of alpha7 nicotinic receptor: cooperative regulation of alpha2-integrin expression and Rho kinase activity.
abstract: Defining the signaling mechanisms and effector proteins mediating phenotypic and mechanical plasticity of keratinocytes (KCs) during wound epithelialization is one of the major goals in epithelial cell biology. The acetylcholine (ACh)-gated ion channels, or nicotinic ACh receptors (nAChRs), mediate the nicotinergic signaling that controls crawling locomotion of KCs. To elucidate relative contributions of the ionic and protein kinase-mediated events elicited due to activation of alpha7 nAChRs, we quantitated expression of alpha2-integrin gene at the mRNA and protein levels and also measured Rho kinase activity in KCs stimulated with the alpha7 agonist AR-R17779 while blocking the Na+ or Ca2+ entry and/or inhibiting signaling kinases. The results demonstrated the existence of the two-component signaling systems coupling the ionic events and protein kinase signaling cascades downstream of alpha7 nAChR to simultaneous up-regulation of alpha2-integrin expression and activation of Rho kinase. The Raf/MEK1/ERK1/2 cascade up-regulating alpha2-integrin was activated due to both Ca2+-dependent recruitment of Ca2+/calmodulin-dependent protein kinase II and protein kinase C and Ca2+-independent activation of Ras. Likewise the phosphatidylinositol 3-kinase-mediated activation of Rho kinase was elicited due to both Ca2+ entry-dependent involvement of Ca2+/calmodulin-dependent protein kinase II and Ca2+-independent activation of Jak2. Thus, although the initial signals emanating from activated alpha7 nAChR are different in nature the pathways intersect at common effector molecules providing for a common end point effect. This novel paradigm of nAChR-mediated coordination of the ionic and metabolic signaling events can allow an auto/paracrine ACh to simultaneously alter gene expression and induce reciprocal changes in the cytoskeleton and contractile system of KCs required to compete a particular step of wound epithelialization.
authors: Departments of Dermatology and Biological Chemistry, University of California, Irvine, California 92697.
authors: Alexander I
authors: AI
authors: Chernyavsky
authors: Departments of Dermatology and Biological Chemistry, University of California, Irvine, California 92697.
authors: Juan
authors: J
authors: Arredondo
authors: Departments of Dermatology and Biological Chemistry, University of California, Irvine, California 92697.
authors: Jing
authors: J
authors: Qian
authors: Departments of Dermatology and Biological Chemistry, University of California, Irvine, California 92697.
authors: Valentin
authors: V
authors: Galitovskiy
authors: Departments of Dermatology and Biological Chemistry, University of California, Irvine, California 92697.
authors: Sergei A
authors: SA
authors: Grando
mesh_terms: D002118:Calcium / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D039421:Integrin alpha2 / Q000096:biosynthesis*; D007477:Ions; D053614:Janus Kinase 2 / Q000378:metabolism; D015603:Keratinocytes / Q000378:metabolism; D008954:Models, Biological; D010641:Phenotype; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D011978:Receptors, Nicotinic / Q000378:metabolism*; D015398:Signal Transduction; D064569:alpha7 Nicotinic Acetylcholine Receptor; D054460:rho-Associated Kinases / Q000378:metabolism*
chemical_list: C579167:Chrna7 protein, human; D039421:Integrin alpha2; D007477:Ions; D011978:Receptors, Nicotinic; D064569:alpha7 Nicotinic Acetylcholine Receptor; D011494:Protein Kinases; C507924:JAK2 protein, human; D053614:Janus Kinase 2; D054460:rho-Associated Kinases; D002118:Calcium