Identification and immunolocalization of chondroitin sulfate proteoglycans in tooth cementum
Identification and immunolocalization of chondroitin sulfate proteoglycans in tooth cementum
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DOI：
10.3109/03008209909005276
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发表时间：
1999-01-01
影响因子：
2.9
通讯作者：
Yamauchi, M
中科院分区：
文献类型：
--
作者：
Cheng, H;Caterson, B;Yamauchi, M
蛋白聚糖（PG）在其核心蛋白以及碳水化合物结构中显示出极大的多样性，并且被认为参与许多生物学功能。最近，我们已经在牛牙骨质中鉴定并免疫定位了两种硫酸角质素PG，纤调蛋白和光蛋白聚糖（Cheng Pt al.，Connect.组织研究34：87-96，1996），本研究的目的是鉴定和表征牙骨质中的硫酸软骨素（CS）PG。为了探索它们与矿物质的潜在关联，通过连续提取将牛牙骨质基质分子分级分离成未结合矿物质的基质和结合矿物质的基质。两种组分均经过DEAE阴离子交换柱层析，收集的样品分别用特异性单克隆抗体2-B-6和3-R-3进行斑点免疫分析，以测定C_4S和C_6S异构体。经生化和免疫化学分析，前者含有核心蛋白聚糖和双糖蛋白聚糖，后者含有多功能蛋白聚糖。牙骨质多功能蛋白聚糖的C6 S与C4 S异构体的比例约为7：1。此外，这些PG免疫定位在牙齿牙骨质和周围使用针对各自的核心蛋白产生的抗体。多功能蛋白聚糖的强烈免疫染色几乎只出现在牙骨质中的牙骨质细胞和牙槽骨中的骨细胞的腔隙中。核心蛋白聚糖的免疫染色主要与牙周膜中的胶原纤维有关，牙骨质基质中的胶原纤维也有少量，而双糖蛋白聚糖的免疫染色主要在成牙骨质细胞/前牙骨质中。这些差异的组织分布的CSPGs表明，他们可能在牙骨质形成中发挥独特的作用。
Proteoglycans (PGs) display a great diversity in their core proteins as well as carbohydrate structures and are thought to be involved in many biological functions, Recently we have identified and immunolocalized two keratan sulfate PGs, fibromodulin and lumican, in bovine tooth cementum (Cheng Pt al,, Connect. Tissue Res. 34: 87-96, 1996), The objectives of this study were to identify and characterize chondroitin sulfate (CS) PGs in cementum, In order to explore their potential association with mineral, bovine cementum matrix molecules were fractionated into mineral-unbound and -bound matrices by sequential extraction. Both fractions were subjected to DEAE anion exchange column chromatography and the eluate collected was assayed for C4S and C6S isomers by dot blot immunoassay with specific monoclonal antibodies, 2-B-6 and 3-R-3, respectively, Two families of CSPGs were identified mainly in the mineral-unbound fraction. One contained only C4S glycosaminoglycan and the other both C6S and C4S, By biochemical and immunochemical analyses, decorin and biglycan were identified in the former and versican in the latter. The ratio of C6S to C4S isomers of cementum versican was approximately 7:1. Furthermore, these PGs were immunolocalized in and around tooth cementum using antibodies generated against the respective core proteins. Intensive immunostaining for versican was found almost exclusively in the lacunae housing cementocytes in cementum and osteocytes in alveolar hone, respectively. Immunostaining for decorin was mainly associated with collagen fibers in the periodontal ligament and slightly in cementum matrix, while the one for biglycan was mainly in cementoblasts/precementum. These differential tissue distributions of the CSPGs suggest that they may play distinct roles in cementogenesis.