A NICOTINIC ACETYLCHOLINE-RECEPTOR REGULATING CELL-ADHESION AND MOTILITY IS EXPRESSED IN HUMAN KERATINOCYTES

被引:177
作者
GRANDO, SA
HORTON, RM
PEREIRA, EFR
DIETHELMOKITA, BM
GEORGE, PM
ALBUQUERQUE, EX
CONTIFINE, BM
机构
[1] UNIV MINNESOTA,SCH MED,DEPT PHARMACOL,MINNEAPOLIS,MN 55455
[2] UNIV MINNESOTA,COLL BIOL SCI,DEPT BIOCHEM,ST PAUL,MN 55108
[3] UNIV MARYLAND,SCH MED,DEPT PHARMACOL & EXPTL THERAPEUT,BALTIMORE,MD 21201
[4] FED UNIV RIO DE JANEIRO,INST BIOFIS CARLOS CHAGAS FILHO,MOLEC PHARMACOL LAB,BR-21945 RIO JANEIRO,BRAZIL
关键词
PATCH CLAMP; RADIOLIGAND BINDING; IMMUNOFLUORESCENCE; POLYMERASE CHAIN REACTION;
D O I
10.1111/1523-1747.ep12325606
中图分类号
R75 [皮肤病学与性病学];
学科分类号
100206 ;
摘要
Acetylcholine is synthesized and released by human epidermal keratinocytes and modulates the adhesion and motility of these cells. To understand the molecular basis of the effects of acetylcholine on keratinocytes, we investigated the presence, pharmacology, structure, and function of nicotinic acetylcholine receptors in human epidermal keratinocytes. Patch-clamp studies indicated that keratinocytes express acetylcholine receptors with ion gating and pharmacologic properties similar to those observed so far only in neurons, and containing the alpha(3) subunit. Specific binding of the receptor-specific ligand I-125-kappa-bungarotoxin revealed approximately 5500 binding sites per cell on undifferentiated keratinocytes in cell cultures and approximately 35,400 binding sites per cell on mature keratinocytes freshly isolated from human neonatal foreskins. Antibody binding and polymerase chain reaction experiments demonstrated the presence of alpha(3), beta(2), and beta(4) nicotinic receptor subunits. Binding of subunit-specific antibodies indicated that nicotinic receptors were associated with the suprabasal keratinocytes in epidermis and localized to the cell membranes of differentiated keratinocytes in cell cultures. Acetylcholine and the nicotinic agonist nicotine increased cell-substrate and cell-cell adherence of cultured keratinocytes and stimulated their lateral migration. The specific antagonists K-bungarotoxin and mecamylamine caused cell detachment and abolished migration. Thus, a nicotinic receptor expressed in keratinocytes may mediate acetylcholine control of keratinocyte adhesion and motility.
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页码:774 / 781
页数:8
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