Regulation of Keratinocyte Migration by Acetylcholine

Summary

Principal Investigator: SERGEI GRANDO
Abstract: Purpose: To integrate structural and functional information about epidermal keratinocytes (KCs) during epithelialization with the knowledge on cholinergic mechanisms guiding KCs through epithelialization steps. Rationale: Epithelialization is a self-regulated process in which cell activities mediating adhesion and migration are controlled, in part, by a single "pace-maker" system featuring autocrine, juxtacrine and paracrine acetylcholine (ACh) as a chemokine for cell motility. Background: KCs are simultaneously stimulated through two distinct types of cholinergic signaling pathways: the metabolic events, elicited by ACh binding to G protein-coupled muscarinic receptors (mAChRs), and the ionic events, mediated by ACh-gated ion channels (or nAChRs). This diversity allows the single cytotransmitter ACh to exert diverse effects on KCs at various stages of skin epithelialization. Working Hypotheses: The MI mAChR and the a3|34 and a7 nAChRs act synergistically in mediating chemotaxis via Ca2+-dependent alterations in the phosphorylation status of cadherins and sedentary integrins. M4 mAChR and a3 nAChR facilitate migration by upregulating migratory integrins through protein kinase (PK) CS-dependent pathway, whereas a9 nAChR prompts assembly/disassemblyof cell-cell and cell- substrate bonds. The aS-containing 3 nAChR increases the strength of cell-cell adhesion. M3 elicits stable cell-substrate attachment and terminates migration due to inactivation of the Rho-dependent events. Specific Aims: 1) Identify ACh receptor subtypes regulating distinct cellular activities of KCs required for normal epithelialization. 2) Identify the ACh receptor-coupled signal transduction pathways mediating selective regulation of cellular activities comprising the keratinocyte migratory function. Methodology: A combination of molecular biological (real-time PCR;small interfering RNA;ACh receptor knockout mice), immunological (immunohistochemistry, Western blot, ELISA, flow cytometry), and pharmacological approaches will be employed to elucidate cholinergic regulation of epithelialization, using the assays of keratinocyte chemotaxis, chemokinesis, and cell-cell and cell-substrate attachments. Significance: Results of this study will lay a groundwork for designing therapeutic regimens able to treat the skin disease caused by abnormal epithelialization. DESCRIPTION: Recent research shows that the locally produced and released hormone-like molecule, ACh, may initiate, guide, and sustain skin cell motility in the wound bed. This makes ACh and drugs acting at the ACh receptors potentially powerful tools for wound healing. This study will identify potential therapeutic agents among cholinergic drugs acting upon specific types of ACh receptors to activate signaling pathways mediating keratinocyte functions during wound epithelialization.
Funding Period: ----------------2000 - ---------------2011-
more information: NIH RePORT

Top Publications

  1. ncbi Cholinergic control of epidermal cohesion
    Sergei A Grando
    Department of Dermatology, University of California at Davis, Sacramento, CA 95817, USA
    Exp Dermatol 15:265-82. 2006
  2. ncbi SLURP-2: A novel cholinergic signaling peptide in human mucocutaneous epithelium
    Juan Arredondo
    Department of Dermatology, University of California, Davis, California, USA
    J Cell Physiol 208:238-45. 2006
  3. ncbi Adrenergic and cholinergic control in the biology of epidermis: physiological and clinical significance
    Sergei A Grando
    Department of Dermatology, University of California, Davis, 95817, USA
    J Invest Dermatol 126:1948-65. 2006
  4. pmc The M4 muscarinic acetylcholine receptor plays a key role in the control of murine hair follicle cycling and pigmentation
    Sybille Hasse
    Department of Dermatology, University Hospital Schleswig Holstein, Campus Luebeck, University of Luebeck, D 23538 Luebeck, Germany
    Life Sci 80:2248-52. 2007
  5. pmc Central role of alpha9 acetylcholine receptor in coordinating keratinocyte adhesion and motility at the initiation of epithelialization
    Alex I Chernyavsky
    Department of Dermatology, University of California Irvine, C340 Medical Sciences I, Irvine, CA 92697, USA
    Exp Cell Res 313:3542-55. 2007
  6. ncbi Differential coupling of M1 muscarinic and alpha7 nicotinic receptors to inhibition of pemphigus acantholysis
    Alex I Chernyavsky
    Department of Dermatology, University of California, Irvine, California 92697, USA
    J Biol Chem 283:3401-8. 2008
  7. pmc Novel cholinergic peptides SLURP-1 and -2 regulate epithelialization of cutaneous and oral wounds
    Alexander I Chernyavsky
    Department of Dermatology, University of California, Irvine, California 92697, USA
    Wound Repair Regen 20:103-13. 2012
  8. pmc Upregulation of nuclear factor-kappaB expression by SLURP-1 is mediated by alpha7-nicotinic acetylcholine receptor and involves both ionic events and activation of protein kinases
    Alexander I Chernyavsky
    Department of Dermatology, University of California, Irvine, California 92697, USA
    Am J Physiol Cell Physiol 299:C903-11. 2010
  9. pmc Antimitochondrial autoantibodies in pemphigus vulgaris: a missing link in disease pathophysiology
    Steve Marchenko
    Department of Dermatology and Biological Chemistry, Institute for Immunology, University of California, Irvine, California 92697, USA
    J Biol Chem 285:3695-704. 2010
  10. pmc Auto/paracrine control of inflammatory cytokines by acetylcholine in macrophage-like U937 cells through nicotinic receptors
    Alexander I Chernyavsky
    Institute for Immunology, University of California, Irvine, CA 92697, USA
    Int Immunopharmacol 10:308-15. 2010

Scientific Experts

  • Daniel K Hsu
  • SERGEI GRANDO
  • Alexander I Chernyavsky
  • Juan Arredondo
  • Alex I Chernyavsky
  • Steve Marchenko
  • Valentin Galitovskiy
  • Jing Qian
  • Sybille Hasse
  • Mina Kalantari-Dehaghi
  • Courtney Phillips
  • Maryna Skok
  • Vivian Gindi
  • Evert Karlsson
  • Timothy Piser
  • Ralf Paus
  • Douglas E Vetter
  • David L Jolkovsky
  • Robert J Webber

Detail Information

Publications13

  1. ncbi Cholinergic control of epidermal cohesion
    Sergei A Grando
    Department of Dermatology, University of California at Davis, Sacramento, CA 95817, USA
    Exp Dermatol 15:265-82. 2006
    ..Recent progress with the development of antiacantholytic therapies of patients with pemphigus using cholinomimetics indicates that cholinergic drugs may be a promising approach for other cutaneous blistering disorders...
  2. ncbi SLURP-2: A novel cholinergic signaling peptide in human mucocutaneous epithelium
    Juan Arredondo
    Department of Dermatology, University of California, Davis, California, USA
    J Cell Physiol 208:238-45. 2006
    ....
  3. ncbi Adrenergic and cholinergic control in the biology of epidermis: physiological and clinical significance
    Sergei A Grando
    Department of Dermatology, University of California, Davis, 95817, USA
    J Invest Dermatol 126:1948-65. 2006
    ..A detailed understanding of the physiology and pathophysiology of the adrenergic/cholinergic network in the skin could offer the development of specific drugs for novel treatment modalities...
  4. pmc The M4 muscarinic acetylcholine receptor plays a key role in the control of murine hair follicle cycling and pigmentation
    Sybille Hasse
    Department of Dermatology, University Hospital Schleswig Holstein, Campus Luebeck, University of Luebeck, D 23538 Luebeck, Germany
    Life Sci 80:2248-52. 2007
    ..The current pilot study suggests that the M4R plays a fundamental role in the control of the murine hair follicle cycling and is an essential signaling element in the control of hair follicle pigmentation...
  5. pmc Central role of alpha9 acetylcholine receptor in coordinating keratinocyte adhesion and motility at the initiation of epithelialization
    Alex I Chernyavsky
    Department of Dermatology, University of California Irvine, C340 Medical Sciences I, Irvine, CA 92697, USA
    Exp Cell Res 313:3542-55. 2007
    ..By activating alpha9 AChR, ACh can control the dynamics and strength of cell-cell cohesion, disabling of a trailing uropod and disassembly and reassembly of focal adhesions, thus facilitating crawling locomotion...
  6. ncbi Differential coupling of M1 muscarinic and alpha7 nicotinic receptors to inhibition of pemphigus acantholysis
    Alex I Chernyavsky
    Department of Dermatology, University of California, Irvine, California 92697, USA
    J Biol Chem 283:3401-8. 2008
    ..These findings identified novel paradigm of regulation of signaling kinases associated with cholinergic receptors and provided mechanistic explanation of therapeutic activity of cholinomimetics in PV patients...
  7. pmc Novel cholinergic peptides SLURP-1 and -2 regulate epithelialization of cutaneous and oral wounds
    Alexander I Chernyavsky
    Department of Dermatology, University of California, Irvine, California 92697, USA
    Wound Repair Regen 20:103-13. 2012
    ..The cholinergic peptides, therefore, may become prototype drugs for the treatment of wounds that fail to heal...
  8. pmc Upregulation of nuclear factor-kappaB expression by SLURP-1 is mediated by alpha7-nicotinic acetylcholine receptor and involves both ionic events and activation of protein kinases
    Alexander I Chernyavsky
    Department of Dermatology, University of California, Irvine, California 92697, USA
    Am J Physiol Cell Physiol 299:C903-11. 2010
    ..Thus, there exists a previously not appreciated network of noncanonical auto/paracrine ligands of nAChR of the Ly-6 protein family, which merits further investigations...
  9. pmc Antimitochondrial autoantibodies in pemphigus vulgaris: a missing link in disease pathophysiology
    Steve Marchenko
    Department of Dermatology and Biological Chemistry, Institute for Immunology, University of California, Irvine, California 92697, USA
    J Biol Chem 285:3695-704. 2010
    ..Further elucidation of the primary signaling events downstream of PV autoantigens will be crucial for the development of a more successful therapy for PV patients...
  10. pmc Auto/paracrine control of inflammatory cytokines by acetylcholine in macrophage-like U937 cells through nicotinic receptors
    Alexander I Chernyavsky
    Institute for Immunology, University of California, Irvine, CA 92697, USA
    Int Immunopharmacol 10:308-15. 2010
    ..These findings offer a new insight on how nicotinic agonists control inflammation, thus laying a groundwork for the development of novel immunomodulatory therapies based on the nAChR subtype selectivity of nicotinic agonists...
  11. pmc 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
    Alexander I Chernyavsky
    Department of Dermatology, University of California, Irvine, California 92697, USA
    J Biol Chem 284:22140-8. 2009
    ....
  12. pmc Endogenous galectin-3 is localized in membrane lipid rafts and regulates migration of dendritic cells
    Daniel K Hsu
    Department of Dermatology, University of California Davis School of Medicine, Sacramento, California 95817, USA
    J Invest Dermatol 129:573-83. 2009
    ..In view of the participation of membrane ruffles in signal transduction and cell motility, we conclude that galectin-3 regulates cell migration by functioning at these structures...
  13. ncbi Muscarinic receptor agonists and antagonists: effects on keratinocyte functions
    Sergei A Grando
    Departments of Dermatology and Biological Chemistry, University of California, 134 Sprague Hall, Irvine, CA, 92697, USA
    Handb Exp Pharmacol 208:429-50. 2012
    ..Successful therapy of pemphigus lesions with topical pilocarpine and disappearance of psoriatic lesions due to systemic atropine therapy illustrate that such therapeutic approach is feasible...