Góes R M, Laicine E M, Porcionatto M A, Bonciani Nader H, Haddad A
UNESP Departamento de Biologia Brasil São José do Rio Preto, SP,
Curr Eye Res. 1999 Aug;19(2):146-53. doi: 10.1076/ceyr.19.2.146.5324.
To trace the eye components involved in proteoglycan synthesis and to characterize the sulfated glycosaminoglycans which are associated to these macromolecules.
Sodium [(35)S]-sulfate was injected intravitreally and the rabbits were killed at different time intervals after the injection. The glycosaminoglycans of choroid, ciliary body, cornea, iris, lens capsule, retina and sclera were extracted and processed for estimations of their specific activities, and for electrophoresis plus autoradiography with or without previous treatment with specific enzymes. In addition, methacrylate sections of the eyes were analysed by autoradiography.
The peak of specific activities of the glycosaminoglycans of all eye components occurred at 2 days after the intravitreal injection of [( 35)S]-sulfate. The autoradiography of the agarose gels revealed three types of glycosaminoglycans, namely, heparan-, chondroitin- and dermatan sulfate, only in the retina. The other eye components contained heparan sulfate and either chondroitin or dermatan sulfate. Tissue autoradiography together with the biochemical techniques contributed to unravel the origin of the glycosaminoglycans in the eye components.
The results of the present investigation have shown that heparan sulfate, contrasting to chondroitin sulfate and dermatan sulfate, is synthesized in all eye components studied and that the glycosaminoglycan composition differs according to the tissue of origin.
追踪参与蛋白聚糖合成的眼部成分,并对与这些大分子相关的硫酸化糖胺聚糖进行表征。
将[35S] - 硫酸钠玻璃体内注射,注射后在不同时间间隔处死兔子。提取脉络膜、睫状体、角膜、虹膜、晶状体囊、视网膜和巩膜的糖胺聚糖,并进行处理以估计其比活性,以及进行电泳和放射自显影,有无用特定酶预先处理。此外,通过放射自显影分析眼睛的甲基丙烯酸酯切片。
玻璃体内注射[35S] - 硫酸钠后2天,所有眼部成分的糖胺聚糖比活性达到峰值。琼脂糖凝胶的放射自显影仅在视网膜中显示出三种类型的糖胺聚糖,即硫酸乙酰肝素、硫酸软骨素和硫酸皮肤素。其他眼部成分含有硫酸乙酰肝素以及硫酸软骨素或硫酸皮肤素。组织放射自显影与生化技术共同有助于揭示眼部成分中糖胺聚糖的来源。
本研究结果表明,与硫酸软骨素和硫酸皮肤素不同,硫酸乙酰肝素在所研究的所有眼部成分中均有合成,并且糖胺聚糖的组成因组织来源而异。