Valiyaveettil Manojkumar, Mort John S, McDevitt Cahir A
Department of Biomedical Engineering, Lerner Research Institute and Orthopaedic Research Center, Cleveland Clinic Foundation, Cleveland, Ohio 44195, USA.
Connect Tissue Res. 2005;46(2):83-91. doi: 10.1080/03008200590954113.
The concentration, spatial distribution, and gene expression of aggrecan in meniscus, articular cartilage, and the anterior and posterior cruciate ligaments (ACL and PCL) was determined in the knee joints of five mature dogs. An anti-serum against peptide sequences specific to the G1 domain of aggrecan was employed in competitive-inhibition ELISA of guanidine HCl extracts and immunofluorescence microscopy. Gene expression was determined by Taqman real-time PCR. The concentration of aggrecan in articular cartilage (240.1 +/- 32 nMol/g dry weight) was higher than that in meniscus (medial meniscus: 33.4 +/- 4.3 nMol/g) and ligaments (ACL: 6.8 +/- 0.9 nMol/g). Aggrecan was more concentrated in the inner than the outer zone of the meniscus. Aggrecan in meniscus showed an organized, spatial network, in contrast to its diffuse distribution in articular cartilage. Thus, differences in the concentration, gene expression, and spatial distribution of aggrecan constitute another molecular distinction between hyaline cartilage and fibrocartilage of the knee.
在五只成年犬的膝关节中,测定了半月板、关节软骨以及前后交叉韧带(前交叉韧带和后交叉韧带)中聚集蛋白聚糖的浓度、空间分布和基因表达。采用针对聚集蛋白聚糖G1结构域特异性肽序列的抗血清,对盐酸胍提取物进行竞争性抑制酶联免疫吸附测定(ELISA)以及免疫荧光显微镜检查。通过Taqman实时聚合酶链反应(PCR)测定基因表达。关节软骨中聚集蛋白聚糖的浓度(240.1±32纳摩尔/克干重)高于半月板(内侧半月板:33.4±4.3纳摩尔/克)和韧带(前交叉韧带:6.8±0.9纳摩尔/克)。聚集蛋白聚糖在半月板内侧比外侧更为集中。半月板中的聚集蛋白聚糖呈现出有组织的空间网络,与其在关节软骨中的弥漫分布形成对比。因此,聚集蛋白聚糖在浓度、基因表达和空间分布上的差异构成了膝关节透明软骨和纤维软骨之间的另一个分子区别。