Franke W W, Schmid E, Grund C, Müller H, Engelbrecht I, Moll R, Stadler J, Jarasch E D
Differentiation. 1981;20(3):217-41. doi: 10.1111/j.1432-0436.1981.tb01178.x.
Desmosome-enriched fractions were isolated from bovine muzzle epidermis either as desmosome-tonofilament complexes using a procedure involving treatment at pH9 or in the form of desmosomal residue fractions using a modification of the citric acid buffer (pH 2.3) method of Skerrow and Matoltsy [1]. Major polypeptides of high molecular weights (mol. wt.) were separated by gel electrophoresis, individual polypeptide bands were excised, and protein was eluted and used for immunization. Guinea pig antibodies raised against two prominent polypeptides of high mol. wt. (250,000 and 215,000) showed, on nitrocellulose paper blots of desmosome-tonofilament polypeptides separated by gel electrophoresis, extensive cross-reaction between a group or large polypeptides characteristic of desmosome-containing fractions, most notably polypeptides of 250 K, 215 K, 200 k, 175 K, and 164 K. These antibodies allowed, when used in immunofluorescence microscopy, the specific localization of desmosomal junctions (i) in sections through epithelia-containing tissue (e.g., epidermis, mucosae of tongue and esophagus, cornea, mammary gland, small intestine, liver, thymus, urothelium of bladder) and myocardium; (ii) on dissociated cells from these tissues; (iii) on various epithelial cells grown in culture; an (iv) in tumor-like proliferations of cultured epithelial cells injected into nude mice. Individual desmosomes could be visualized and resolved at the light microscopic level. No reaction was found in cells devoid of desmosomes and on other classes of intercellular junctions. Electron microscopic localization using immunoperoxidase techniques indicated that these proteins are located in, or close to, the desmosomal plague structure. It is proposed to use such antibodies against desmosomal proteins as markers specific to this so far only morphologically define class of junctions. Use of these markers will (i) improve identification and classification of intercellular junctions; (ii) facilitate determinations of the specific patterns of distributions of desmosomes and desmosomal protein in various cells and tissue; (iii) allow studies of formation and disintegration of desmosomes, and of the biosynthesis and possible recycling of their constituents; and (iv) provide tissue group-specific markers valuable in histology and diagnosis, especially for identification of epithelial and carcinoma cells.
富含桥粒的组分从牛口鼻部表皮中分离得到，分离方法有两种：一种是采用pH9处理的程序，将其作为桥粒-张力丝复合体进行分离；另一种是采用对Skerrow和Matoltsy [1]的柠檬酸缓冲液（pH 2.3）方法进行改良，以桥粒残余物组分的形式进行分离。通过凝胶电泳分离出高分子量（mol. wt.）的主要多肽，切下各个多肽条带，洗脱蛋白质并用于免疫。针对两种高分子量（250,000和215,000）的突出多肽产生的豚鼠抗体，在经凝胶电泳分离的桥粒-张力丝多肽的硝酸纤维素纸印迹上显示，在含桥粒组分特有的一组或大分子量多肽之间存在广泛的交叉反应，最显著的是250K、215K、200k、175K和16aK的多肽。当用于免疫荧光显微镜检查时，这些抗体能够特异性定位桥粒连接：（i）在含有上皮组织（如表皮、舌和食管黏膜、角膜、乳腺、小肠、肝脏、胸腺、膀胱尿路上皮）和心肌的切片中；（ii）在来自这些组织的解离细胞上；（iii）在培养的各种上皮细胞上；以及（iv）在注射到裸鼠体内的培养上皮细胞的肿瘤样增殖物中。在光学显微镜水平可以观察到并分辨出单个桥粒。在没有桥粒的细胞和其他类型的细胞间连接上未发现反应。使用免疫过氧化物酶技术进行电子显微镜定位表明，这些蛋白质位于桥粒斑结构内或附近。建议使用针对桥粒蛋白的此类抗体作为迄今为止仅在形态学上定义的这种连接类型的特异性标记物。使用这些标记物将：（i）改进细胞间连接的鉴定和分类；（ii）便于确定桥粒和桥粒蛋白在各种细胞和组织中的特定分布模式；（iii）允许研究桥粒的形成和分解及其成分的生物合成和可能的再循环；以及（iv）提供在组织学和诊断中具有重要价值的组织组特异性标记物，特别是用于鉴定上皮细胞和癌细胞。
