Nishikawa S, Sasaki F
Department of Biology, Tsurumi University, School of Dental Medicine, Yokohama, Japan.
J Histochem Cytochem. 1993 Sep;41(9):1373-81. doi: 10.1177/41.9.8354877.
We examined proteoglycans (PGs) in amphibian epidermal cells by immunofluorescence microscopy. Immunoelectron microscopy with pre- and post-embedding methods, combined with HRP- or gold-conjugated secondary antibody, revealed ultrastructural localization of glycosaminoglycans (GAGs). Embryonic epidermis secretion granules in Xenopus laevis contained chondroitin 6-sulfate and unsulfated chondroitin. Immature secretion granules were also labeled with anti-chondroitin 6-sulfate and anti-chondroitin. A step-wise digestion experiment on chondroitinase ABC revealed that fine filaments in the secretion granules were chondroitin sulfate chains. Inhibition experiments with brefeldin A revealed that the life of a secretion granule was 5-10 hr, suggesting that GAG secretion from embryonic epidermis is routed through a regulated pathway.
我们通过免疫荧光显微镜检查了两栖动物表皮细胞中的蛋白聚糖（PGs）。采用包埋前和包埋后方法的免疫电子显微镜，结合辣根过氧化物酶（HRP）或金标记二抗，揭示了糖胺聚糖（GAGs）的超微结构定位。非洲爪蟾胚胎表皮分泌颗粒含有硫酸软骨素6和未硫酸化的软骨素。未成熟的分泌颗粒也用抗硫酸软骨素6和抗软骨素标记。对软骨素酶ABC进行的逐步消化实验表明，分泌颗粒中的细丝是硫酸软骨素链。用布雷菲德菌素A进行的抑制实验表明，分泌颗粒的寿命为5至10小时，这表明胚胎表皮中GAG的分泌是通过一条受调控的途径进行的。
