title: Choline acetyltransferase, acetylcholinesterase, and nicotinic acetylcholine receptors of human gingival and esophageal epithelia.
abstract: A non-neuronal cholinergic system that includes neuronal-like nicotinic acetylcholine receptors (nAChRs) has recently been described in epithelial cells that line the skin and the upper respiratory tract. Since the use of nicotine-containing products is associated with morbidity in the upper digestive tract, and since nicotine may alter cellular functions directly via nAChRs, we sought to identify and characterize a non-neuronal cholinergic system in the gingival and esophageal epithelia. mRNA transcripts for alpha3, alpha5, alpha7, and beta2 nAChR subunits, choline acetyltransferase, and the asymmetric and globular forms of acetylcholinesterase were amplified from gingival keratinocytes (KC) by means of polymerase chain-reactions. These proteins were visualized in the gingival and esophageal epithelia by means of specific antibodies. Variations in distribution and intensity of immunostaining were found, indicating that the repertoire of cholinergic enzymes and receptors expressed by the cells changes during epithelial maturation, and that an upward concentration gradient of free acetylcholine exists. Blocking of the nAChRs with mecamylamine resulted in reversible loss of cell-to-cell adhesion, and shrinking and rounding of cultured gingival KC. Activation of the receptors with acetylcholine or carbachol caused stretching and peripheral ruffling of the cytoplasmic aprons, and formation of new intercellular contacts. These results demonstrate that both the keratinizing epithelium of attached gingiva and the non-keratinizing epithelium lining the upper two-thirds of the esophageal mucosa possess a non-neuronal cholinergic system. The nAChRs expressed by these epithelia are coupled to regulation of cell adhesion and motility, and may provide a target for the deleterious effects of nicotine.
authors.affiliation: Department of Dermatology, University of California-Davis, UC Davis Medical Center, Sacramento 95817, USA.
authors.forename: V T
authors.initials: VT
authors.lastname: Nguyen
authors.forename: L L
authors.initials: LL
authors.lastname: Hall
authors.forename: G
authors.initials: G
authors.lastname: Gallacher
authors.forename: A
authors.initials: A
authors.lastname: Ndoye
authors.forename: D L
authors.initials: DL
authors.lastname: Jolkovsky
authors.forename: R J
authors.initials: RJ
authors.lastname: Webber
authors.forename: R
authors.initials: R
authors.lastname: Buchli
authors.forename: S A
authors.initials: SA
authors.lastname: Grando
mesh_terms: D000109:Acetylcholine / Q000494:pharmacology; D000110:Acetylcholinesterase / Q000032:analysis* / Q000235:genetics; D000906:Antibodies; D002217:Carbachol / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D002478:Cells, Cultured; D002795:Choline O-Acetyltransferase / Q000032:analysis* / Q000037:antagonists & inhibitors / Q000235:genetics; D018679:Cholinergic Agonists / Q000494:pharmacology; D002800:Cholinesterase Inhibitors / Q000494:pharmacology; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D004947:Esophagus / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D005881:Gingiva / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008464:Mecamylamine / Q000494:pharmacology; D009092:Mucous Membrane / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D009538:Nicotine / Q000009:adverse effects; D018722:Nicotinic Agonists / Q000009:adverse effects; D018733:Nicotinic Antagonists / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics; D011978:Receptors, Nicotinic / Q000032:analysis* / Q000235:genetics
chemical_list: D000906:Antibodies; D018679:Cholinergic Agonists; D002800:Cholinesterase Inhibitors; D018722:Nicotinic Agonists; D018733:Nicotinic Antagonists; D012333:RNA, Messenger; D011978:Receptors, Nicotinic; D008464:Mecamylamine; D009538:Nicotine; D002217:Carbachol; D002795:Choline O-Acetyltransferase; D000110:Acetylcholinesterase; D000109:Acetylcholine