Uronic acid-containing glycosaminoglycans and keratan sulfate are present in the tectorial membrane of the inner ear: functional implications.
Uronic acid-containing glycosaminoglycans and keratan sulfate are present in the tectorial membrane of the inner ear: functional implications.
复制标题
内耳盖膜中存在含糖醛酸的糖胺聚糖和硫酸角质素：功能意义。
DOI：
10.1006/abbi.1993.1605
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发表时间：
1993
影响因子：
3.9
通讯作者：
Thalmann,R
中科院分区：
文献类型：
--
作者：
Thalmann,I;Machiki,K;Calabro,A;Hascall,VC;Thalmann,R
盖膜是一种凝胶状的无细胞结缔组织，覆盖在显微镜下的Corti器官-听觉感觉结构上。它在毛细胞静纤毛的声音同步偏转中起作用，这是听觉传导的中心事件。已经确定，胶原蛋白，主要是II型，构成了覆膜的主要蛋白质，也存在少量的IX和XI型。然而，在这种结构中的蛋白聚糖的结论性信息是缺乏的。用4 M盐酸胍溶剂提取覆膜，并在乙醇沉淀和胶原酶处理后分离蛋白聚糖。基于阳离子染料番红O与糖胺聚糖结合的比色测定，结合酶促技术，检测到大量的硫酸软骨素和硫酸角质素（分别为0.29和0.17%，以湿重计）。软骨素酶消化的样品的琼脂糖-聚丙烯酰胺电泳揭示了一种核心蛋白，其分子量与大软骨蛋白聚糖聚集蛋白聚糖的分子量相似。该蛋白聚糖与抗体3-B-3（识别修饰的软骨素6-硫酸酯键合区寡糖）反应。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳显示了几种与5-D-4反应的低分子量蛋白质，这些蛋白质对硫酸角质素具有特异性，其中一种表现出纤维调蛋白的特征。比较定量方面的各种结缔组织成分的tectonic膜与其他II型胶原蛋白的结构显示，这种组织类似于高度水合软骨。
The tectorial membrane is a gel-like, acellular connective tissue overlying the microscopic organ of Corti - the auditory sensory structure. It is instrumental in the sound-synchronous deflection of the stereocilia of the hair cells, a central event in auditory transduction. It is well established that collagen, primarily type II, constitutes the major protein of the tectorial membrane, with smaller amounts of types IX and XI also present. However, conclusive information on the proteoglycans in this structure is lacking. Tectorial membranes were extracted with a 4 M guanidine-HCl solvent, and proteoglycans isolated after ethanol precipitation and collagenase treatment. A colorimetric assay based on the binding of the cationic dye safranin O to glycosaminoglycans, in combination with enzymatic techniques, detected significant amounts of chondroitin sulfate and keratan sulfate (0.29 and 0.17% on a wet weight basis, respectively). Agarose-polyacrylamide electrophoresis of chondroitinase-digested samples revealed a core protein with a similar molecular mass to that of the large cartilage proteoglycan aggrecan. This proteoglycan reacted with the antibody 3-B-3 (recognizing modified chondroitin 6-sulfate linkage region oligosaccharides). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed several low molecular mass proteins which reacted with 5-D-4, specific for keratan sulfate, one of which showed characteristics of fibromodulin. Comparison of the quantitative aspects of various connective tissue components of tectonal membrane with other type II collagen-containing structures revealed that this tissue resembles highly hydrated cartilage.