Yamada Shuhei, Morimoto Hideto, Fujisawa Toshitaka, Sugahara Kazuyuki
Department of Biochemistry, Kobe Pharmaceutical University, Higashinada-ku, Kobe 658-8558, Japan.
Glycobiology. 2007 Aug;17(8):886-94. doi: 10.1093/glycob/cwm051. Epub 2007 May 19.
The hydrozoan is the simplest organism whose movements are governed by the neuromuscular system, and its de novo morphogenesis can be easily induced by the removal of body parts. These features make the hydrozoan an excellent model for studying the regeneration of tissues in vivo, especially in the nervous system. Although glycosaminoglycans (GAGs) and proteoglycans (PGs) have been implicated in the signaling functions of various growth factors and play critical roles in the development of the central nervous system, the isolation and characterization of GAGs from hydrozoans have never been reported. Here, we characterized GAGs of Hydra magnipapillata. Immunostaining using anti-GAG antibodies showed chondroitin or chondroitin sulfate (CS) in the developing nematocyst, which is a sting organelle specific to cnidarians. The CS-PGs might furnish an environment for assembling nematocyst components, and might themselves be components of nematocysts. Therefore, GAGs were isolated from Hydra and their structural features were examined. A considerable amount of CS, three orders of magnitude less heparan sulfate (HS), but no hyaluronan were found, as in Caenorhabditis elegans. Analysis of the disaccharide composition of HS revealed glucosamine 2-N-sulfation, glucosamine 6-O-sulfation, and uronate 2-O-sulfation. CS contains not only nonsulfated and 4-O-sulfated N-acetylgalactosamine (GalNAc) but also 6-O-sulfated GalNAc. The average molecular size of CS and HS was 110 and 10 kDa, respectively. It has also been established here that CS chains are synthesized on the core protein through the ubiquitous linkage region tetrasaccharide, suggesting that indispensable functions of the linkage region in the synthesis of GAGs have been conserved during evolution.
水螅虫是最简单的由神经肌肉系统控制运动的生物体,其从头形态发生可以通过去除身体部分轻易诱导。这些特征使水螅虫成为研究体内组织再生,尤其是神经系统再生的优秀模型。尽管糖胺聚糖(GAGs)和蛋白聚糖(PGs)已被认为参与各种生长因子的信号传导功能,并在中枢神经系统发育中起关键作用,但从未有过从水螅虫中分离和表征GAGs的报道。在这里,我们表征了巨型乳头水螅的GAGs。使用抗GAG抗体的免疫染色显示,在发育中的刺丝囊中有软骨素或硫酸软骨素(CS),刺丝囊是刺胞动物特有的刺器官。CS-PGs可能为组装刺丝囊成分提供一个环境,并且它们自身可能是刺丝囊的成分。因此,从水螅中分离出GAGs并检查了它们的结构特征。与秀丽隐杆线虫一样,发现了大量的CS,硫酸乙酰肝素(HS)的量少三个数量级,但没有透明质酸。对HS二糖组成的分析揭示了2-N-硫酸化葡糖胺、6-O-硫酸化葡糖胺和2-O-硫酸化糖醛酸。CS不仅含有非硫酸化和4-O-硫酸化的N-乙酰半乳糖胺(GalNAc),还含有6-O-硫酸化的GalNAc。CS和HS的平均分子大小分别为110和10 kDa。这里还确定CS链是通过普遍存在的连接区四糖在核心蛋白上合成的,这表明连接区在GAGs合成中的不可或缺的功能在进化过程中得到了保留。