The distribution of aggregating proteoglycans in articular cartilage: comparison of quantitative immunoelectron microscopy with radioimmunoassay and biochemical analysis.
The distribution of aggregating proteoglycans in articular cartilage: comparison of quantitative immunoelectron microscopy with radioimmunoassay and biochemical analysis.
复制标题
关节软骨中聚集蛋白聚糖的分布：定量免疫电子显微镜与放射免疫分析和生化分析的比较。
DOI：
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发表时间：
1984
影响因子：
3.2
通讯作者：
T. Hardingham
中科院分区：
文献类型：
--
作者：
A. Ratcliffe;P. Fryer;T. Hardingham
电子显微镜免疫定位和放射免疫测定已被用来确定关节软骨中的蛋白多糖的透明质酸结合区的深度的变化。将软骨从关节面到骨缘连续切片，从每个切片中提取蛋白多糖，并使用针对蛋白多糖结合区的抗体通过放射免疫法测定。蛋白多糖被发现是最丰富的中间区和关节面附近的丰富程度最低。在相同的提取物中的己糖醛酸的生化分析表明，与这些和其他已发表的结果一致的蛋白聚糖的分布。结合区抗血清用于Lowicryl K4M树脂包埋的软骨切片的蛋白多糖的电镜免疫定位。用软骨素酶ABC消化切片后，用抗血清和蛋白A包被的金颗粒作为免疫标记定位蛋白聚糖。使用Magiscan图像分析系统定量标记的密度。在整个深度的软骨基质标记是更高的细胞间区域相比，在细胞周围区域，并与深度的免疫标记的量的变化被发现表现出良好的相关性与放射免疫测定的结果。细胞内标记的蛋白多糖主要发现在高尔基体区域和膜结合（分泌）囊泡。
Electron microscopic immunolocalization and radioimmunoassay have been used to determine the variation with depth of the hyaluronate-binding region of proteoglycan in articular cartilage. The cartilage was cut into serial sections from the articular surface to the bony margin, the proteoglycans were extracted from each section and determined by radioimmunoassay using antibodies raised against proteoglycan binding region. Proteoglycans were found to be most abundant in the middle zone and least abundant near the articular surface. Biochemical analysis for hexuronate in the same extracts showed a distribution of proteoglycan in agreement with these and other published results. The binding region antiserum was used for electron microscopic immunolocalization of proteoglycan with ultrathin sections of cartilage embedded in Lowicryl K4M resin. After digestion of the sections with chondroitinase ABC, the proteoglycans were localized using the antiserum and protein A-coated gold particles as immunolabel. The density of labeling was quantified using a Magiscan image analysis system. Throughout the depth of the cartilage matrix labeling was higher in the pericellular regions compared to the intercellular regions, and variation of the amount of immunolabel with depth was found to show a good correlation with the results from radioimmunoassay. Intracellular labeling of proteoglycans was mainly found over the Golgi region and in membrane-bound (secretory) vesicles.
DOI：
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发表时间：
1980
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Poole,AR;Reiner,A;Tang,LH;Rosenberg,L
通讯作者：
Rosenberg,L