Expression of μ-opiate receptor in human epidermis and keratinocytes
Expression of μ-opiate receptor in human epidermis and keratinocytes
复制标题
DOI：
10.1046/j.1523-1747.1998.00259.x
复制
发表时间：
1998-08-01
影响因子：
6.5
通讯作者：
Rufli, T
中科院分区：
文献类型：
--
作者：
Bigliardi, PL;Mei, BQ;Rufli, T
越来越多的证据表明神经递质在皮肤生理学和病理学中起着至关重要的作用。人表皮中阿黑皮素原分子如β-内啡肽的表达和产生表明阿片受体存在于角质形成细胞中。在本文中，我们表明，人表皮角质形成细胞表达的μ阿片受体的mRNA水平和蛋白质水平。用人表皮角质形成细胞的cDNA文库进行聚合酶链反应，得到预期长度的聚合酶链反应产物，通过Southern印迹分析证实其为μ-阿片受体。用μ阿片受体特异性探针原位杂交技术检测了人表皮μ阿片受体。结果表明，μ阿片受体在表皮各层均有胞质表达，以基底上层表达最明显。使用μ阿片受体特异性抗体的免疫组织化学表明表皮也表达蛋白质，并且蛋白质水平在基底层中更高。皮肤中mRNA和蛋白质表达的位置之间的相关性表明μ-阿片受体不仅被转录，而且具有特定的功能。为了证明受体的功能，我们使用来自人皮肤移植物的皮肤器官培养物进行了功能测定。与纳洛酮或β-内啡肽孵育48小时后，表皮中μ阿片受体的表达与对照相比显著下调。这些结果表明，在人表皮中确实存在功能性受体。
There is increasing evidence that neurotransmitters play a crucial role in skin physiology and pathology, The expression and production of proopiomelanocortin molecules such as beta-endorphin in human epidermis suggest that an opiate receptor is present in keratinocytes. In this paper we show that human epidermal keratinocytes express a mu-opiate receptor on both the mRNA level and the protein level. Performing polymerase chain reaction with cDNA libraries from human epidermal keratinocytes gave the polymerase chain reaction products of the expected length, which were confirmed as mu-opiate receptors by Southern blot analysis. Using in situ hybridization techniques with a specific probe for mu-opiate receptors we detected the receptor in human epidermis, There was a cytoplasmic expression in all layers of the epidermis, which was more distinct in the suprabasal layers. Immunohistochemistry using the mu-opiate receptor-specific antibody indicates that epidermis expresses protein as well, and that the protein level is more elevated in the basal layer. The correlation between the locations of both mRNA and protein expression in skin indicates that the mu-opiate receptor has not only been transcribed but also has a specific function, To prove a function of the receptor we performed a functional assay using skin organ cultures from human skin transplants. After 48 h incubation with Naloxone or beta-endorphin the expression of the mu-opiate receptor in epidermis was significantly downregulated compared with the control. These results show that a functional receptor indeed exists in human epidermis.