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健康犬皮肤中表皮角化蛋白丝聚合蛋白和桥粒芯糖蛋白的精细化免疫化学特性研究。

Refined Immunochemical Characterization in Healthy Dog Skin of the Epidermal Cornification Proteins, Filaggrin, and Corneodesmosin.

机构信息

University of Lyon, VetAgro Sup, UP Interaction Cellules Environnement, Marcy l'Etoile, France.

Synelvia, Labège, France.

出版信息

J Histochem Cytochem. 2019 Feb;67(2):85-97. doi: 10.1369/0022155418798807. Epub 2018 Sep 10.

Abstract

Filaggrin (FLG) and corneodesmosin (CDSN) are two key proteins of the human epidermis. FLG loss-of-function mutations are the strongest genetic risk factors for human atopic dermatitis. Studies of the epidermal distribution of canine FLG and CDSN are limited. Our aim was to better characterize the distribution of FLG and CDSN in canine skin. Using immunohistochemistry on beagle skin, we screened a series of monoclonal antibodies (mAbs) specific for human FLG and CDSN. The cross-reactive mAbs were further used using immunoelectron microscopy and Western blotting. The structure of canine CDSN and FLG was determined using publicly available databases. In the epidermis, four anti-FLG mAbs stained keratohyalin granules in the granular keratinocytes and corneocyte matrix of the lower cornified layer. In urea-extracts of dog epidermis, several bands corresponding to proFLG and FLG monomers were detected. One anti-CDSN mAb stained the cytoplasm of granular keratinocytes and cells of both the inner root sheath and medulla of hair follicles. Dog CDSN was located in lamellar bodies, in the extracellular parts of desmosomes and in corneodesmosomes. A protein of 52 kDa was immunodetected. Genomic DNA analysis revealed that the amino acid sequence and structure of canine and human CDSN were highly similar.

摘要

丝聚蛋白(FLG)和角蛋白中间丝相关糖蛋白(CDSN)是人类表皮的两种关键蛋白。FLG 功能丧失突变是人类特应性皮炎最强的遗传风险因素。关于犬类 FLG 和 CDSN 的表皮分布研究有限。我们的目的是更好地描述犬类皮肤中 FLG 和 CDSN 的分布。我们使用针对犬 FLG 和 CDSN 的一系列单克隆抗体(mAb),通过免疫组化对比格犬皮肤进行筛选。这些交叉反应的 mAb 进一步用于免疫电子显微镜和 Western blot 分析。使用公开可用的数据库确定了犬 CDSN 和 FLG 的结构。在表皮中,四种抗 FLG mAb 可在颗粒状角质形成细胞的角质透明颗粒和下部角质化层的角蛋白细胞基质中染色。在犬类表皮的尿素提取物中,检测到几个与 proFLG 和 FLG 单体相对应的条带。一种抗 CDSN mAb 可染色颗粒状角质形成细胞的细胞质以及毛囊内根鞘和髓质的细胞。犬 CDSN 定位于板层小体、桥粒的细胞外部分以及角蛋白桥粒中。检测到 52 kDa 的蛋白质。基因组 DNA 分析表明,犬和人类 CDSN 的氨基酸序列和结构高度相似。

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