A NICOTINIC ACETYLCHOLINE-RECEPTOR REGULATING CELL-ADHESION AND MOTILITY IS EXPRESSED IN HUMAN KERATINOCYTES
A NICOTINIC ACETYLCHOLINE-RECEPTOR REGULATING CELL-ADHESION AND MOTILITY IS EXPRESSED IN HUMAN KERATINOCYTES
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DOI：
10.1111/1523-1747.ep12325606
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发表时间：
1995-12-01
影响因子：
6.5
通讯作者：
CONTIFINE, BM
中科院分区：
文献类型：
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作者：
GRANDO, SA;HORTON, RM;CONTIFINE, BM
乙酰胆碱由人表皮角质形成细胞合成和释放，并调节这些细胞的粘附和运动。为了了解乙酰胆碱对角质形成细胞影响的分子基础，我们研究了人表皮角质形成细胞中烟碱乙酰胆碱受体的存在、药理学、结构和功能。膜片钳研究表明，角质形成细胞表达乙酰胆碱受体，其离子门控和药理学特性与迄今为止仅在神经元中观察到的相似，并含有α（3）亚基。受体特异性配体I-125-κ-银环蛇毒素的特异性结合揭示了细胞培养物中未分化的角质形成细胞上的每个细胞约5500个结合位点，以及新鲜分离自人新生儿包皮的成熟角质形成细胞上的每个细胞约35，400个结合位点。抗体结合和聚合酶链反应实验证明存在α（3）、β（2）和β（4）烟碱受体亚单位。亚单位特异性抗体的结合表明，烟碱受体与表皮中的基底上角质形成细胞相关，并定位于细胞培养物中分化的角质形成细胞的细胞膜。乙酰胆碱和烟碱激动剂尼古丁增加细胞基质和细胞细胞粘附培养角质形成细胞，并刺激他们的侧向迁移。特异性拮抗剂K-银环蛇毒素和美加明引起细胞脱离和废除迁移。因此，角质形成细胞中表达的烟碱受体可能介导乙酰胆碱对角质形成细胞粘附和运动的控制。
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.