Braunewell K H, Martini R, LeBaron R, Kresse H, Faissner A, Schmitz B, Schachner M
Department of Neurobiology, Swiss Federal Institute of Technology, Hönggerberg, Zürich.
Eur J Neurosci. 1995 Apr 1;7(4):792-804. doi: 10.1111/j.1460-9568.1995.tb00682.x.
After transection of adult mouse sciatic nerve, the expression of a chondroitin sulphate epitope recognized by the monoclonal antibody 473-HD (mAb 473-HD) was found to be up-regulated. The epitope was localized immunocytochemically mainly in Schwann cell basal laminae and, more weakly, also in the endoneurium. In cultures of mouse dorsal root ganglion cells, Schwann cells expressed high levels but fibroblasts only low levels of the epitope. To identify the molecule(s) carrying this chondroitin sulphate epitope, human sciatic nerves were extracted with phosphate-buffered saline and shown to contain two chondroitin sulphate proteoglycans of apparent molecular weights of 130 and 900 kDa. The 900 kDa and, more weakly, the 130 kDa proteoglycan were reactive with mAb 473-HD, which was found to recognize chondroitin-6-sulphate as epitope. Following chondroitinase ABC treatment of the 130 kDa proteoglycan, a core protein of approximately 45 kDa was seen and shown to react with polyclonal antibodies against the chondroitin-dermatan sulphate proteoglycan decorin from human fibroblasts. Chondroitinase ABC treatment of the 900 kDa proteoglycan yielded a core protein with a molecular weight of approximately 400 kDa that was recognized by polyclonal antibodies against recombinantly expressed fusion proteins from human versican. After transection of adult mouse sciatic nerves, the distal nerve stumps showed up-regulation of the chondroitin-6-sulphate epitope of the 900 kDa proteoglycan, whereas the core protein of this proteoglycan did not show any detectable change in the level of expression. In contrast, the core protein of the 130 kDa proteoglycan was up-regulated in expression. These observations suggest that versican- and decorin-like molecules may contribute to successful regeneration in the peripheral nervous system of mammals.
在成年小鼠坐骨神经横断后，发现单克隆抗体473-HD（mAb 473-HD）识别的硫酸软骨素表位的表达上调。该表位经免疫细胞化学定位主要在施万细胞基膜中，在内皮中表达较弱。在小鼠背根神经节细胞培养物中，施万细胞高水平表达该表位，而成纤维细胞仅低水平表达。为鉴定携带该硫酸软骨素表位的分子，用人坐骨神经用磷酸盐缓冲盐水提取，结果显示含有两种表观分子量分别为130 kDa和900 kDa的硫酸软骨素蛋白聚糖。900 kDa的蛋白聚糖以及较弱的130 kDa的蛋白聚糖与mAb 473-HD反应，发现该抗体识别硫酸软骨素-6-硫酸酯作为表位。用软骨素酶ABC处理130 kDa的蛋白聚糖后，可见一条约45 kDa的核心蛋白，并显示其与抗人成纤维细胞硫酸软骨素-硫酸皮肤素蛋白聚糖核心蛋白聚糖的多克隆抗体反应。用软骨素酶ABC处理900 kDa的蛋白聚糖产生一条分子量约为400 kDa的核心蛋白，该核心蛋白被抗人多功能蛋白聚糖重组表达融合蛋白的多克隆抗体识别。成年小鼠坐骨神经横断后，远端神经残端显示900 kDa蛋白聚糖的硫酸软骨素-6-硫酸酯表位上调，而该蛋白聚糖的核心蛋白表达水平未显示任何可检测到的变化。相反，130 kDa蛋白聚糖的核心蛋白表达上调。这些观察结果表明，多功能蛋白聚糖样分子和核心蛋白聚糖样分子可能有助于哺乳动物外周神经系统的成功再生。
