Asher R A, Scheibe R J, Keiser H D, Bignami A
Department of Pathology, Harvard Medical School, Boston, Massachusetts, USA.
Glia. 1995 Apr;13(4):294-308. doi: 10.1002/glia.440130406.
Monoclonal antibodies (mAbs) against the major constituents of cartilage extracellular matrix, aggrecan and link protein, were screened by indirect immunofluorescence on frozen sections of bovine spinal cord. Antibodies against aggrecan and link protein gave rise to very similar perineuronal labeling in spinal cord gray matter. Aggrecan and link protein reactivities were seen in other regions of the central nervous system (CNS), although their distributions were not always coincident. Pretreatment of the tissue section with Streptomyces hyaluronidase, which is hyaluronate-specific, led to the loss of both reactivities. On Western blots, anti-aggrecan mAbs reacted with a large chondroitin sulfate proteoglycan. The chondroitinase-treated CNS proteoglycan co-migrated with the chondroitinase- and keratanase-treated cartilage proteoglycan. In CNS tissue homogenates, the addition of Streptomyces hyaluronidase brought about the release of the proteoglycan from the tissue. Anti-link protein mAbs were reactive with two species in the bovine CNS, the mobilities of which were very similar to those of the cartilage link proteins. The release of these species from the tissue required hyaluronidase. A rabbit antiserum against aggrecan was used to identify a similar proteoglycan in the rat CNS. In spinal cord-derived cell cultures, the labeled material was associated with astrocytes. An aggrecan cDNA hybridized to a 9.5 kb mRNA in the rat CNS. We conclude that the perineuronal matrix consists, in part, of a hyaluronate-bound aggrecan-like proteoglycan and link proteins, and that the former is produced by astrocytes.
通过对牛脊髓冰冻切片进行间接免疫荧光法，筛选出了针对软骨细胞外基质主要成分聚集蛋白聚糖和连接蛋白的单克隆抗体（mAb）。针对聚集蛋白聚糖和连接蛋白的抗体在脊髓灰质中产生了非常相似的神经元周围标记。在中枢神经系统（CNS）的其他区域也观察到了聚集蛋白聚糖和连接蛋白的反应性，尽管它们的分布并不总是一致。用特异性作用于透明质酸的透明质酸酶对组织切片进行预处理，导致两种反应性均丧失。在蛋白质印迹法中，抗聚集蛋白聚糖单克隆抗体与一种大的硫酸软骨素蛋白聚糖发生反应。经软骨素酶处理的中枢神经系统蛋白聚糖与经软骨素酶和角蛋白酶处理的软骨蛋白聚糖共同迁移。在中枢神经系统组织匀浆中，添加透明质酸酶会使蛋白聚糖从组织中释放出来。抗连接蛋白单克隆抗体与牛中枢神经系统中的两种蛋白发生反应，它们的迁移率与软骨连接蛋白的迁移率非常相似。这些蛋白从组织中的释放需要透明质酸酶。使用兔抗聚集蛋白聚糖抗血清来鉴定大鼠中枢神经系统中类似的蛋白聚糖。在脊髓来源的细胞培养物中，标记物质与星形胶质细胞相关。一个聚集蛋白聚糖cDNA与大鼠中枢神经系统中的一个9.5 kb mRNA杂交。我们得出结论，神经元周围基质部分由结合透明质酸的聚集蛋白聚糖样蛋白聚糖和连接蛋白组成，并且前者由星形胶质细胞产生。
