GLYCOBIOLOGICAL STUDY OF TREATMENT OF RETINAL DISEASES

视网膜疾病治疗的糖生物学研究

基本信息

批准号：
11671747
负责人：
UEHARA Fumiyuki
金额：
$ 2.3万
依托单位：
KAGOSHIMA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671747/
关键词：
retina retinal degeneration lectin rat galectin-3 sialidase mucin cDNA 細胞

项目摘要

Basic studies for treatments of retinitis pigmentosa and retinoblastoma were performed using rats with constant light-exposure and culture cells of retinoblastoma, respectively. (1) Jacalin (specific for mucin type carbohydrates), galectin-3 (an endogenous animal lectin), and neuraminidase-inhibitor (2,3-dehydro-2-deoxy-N- acetylneuraminic acid) inhibited the process of photoreceptor cell-degeneration. Galectin-3 was considered to play anti-apoptotic role in the retina through photoreceptor cell-degeneration. (2) cDNAs of human and rat core proteins of mucinlike glycoprotein associated with photoreceptor cells (MLGAPC) were isolated using anti-bovine core protein antibody. The MLGAPC was detected not only in the interphotoreceptor matrix but also in the synaptic cleft matrix. By examining the expression of the MLGAPC in developing rat retinas, we determined that the low and high molecular types corresponded to cone and rod types, respectively. Since the mRNA-expression of the MLGAPC decreased after the maturation of the photoreceptor cells, the turnover-rate of MLGAPC was considered to be slow. This feature of the protein is convenient for the MLGAPC to maintain a stable structure of the photoreceptor and outer synaptic layers. (3) The cDNA of human core protein of MLGAPC was introduced into Y79 retinoblastoma cells. Lectin-blot analysis revealed that they produced rod-type MLGAPC with terminal sialic acids. Since no morphological change of the culture cells was observed after the addition of hyaluronan into the cultire medium, the interaction between MLGAPC and hyaluronan was considered to be insufficient for cellular adhesion.

分别使用恒定的光暴露和视网膜细胞瘤的培养细胞进行了色素性视网膜炎和视网膜细胞瘤治疗的基础研究。（1）黑甲林（特异性粘蛋白类型碳水化合物），lectectin-3（内源性动物凝集素）和神经氨酸酶抑制剂（2,3-脱氧-2-脱氧-N-乙酰神经氨酸）抑制了光感受器细胞基因化的过程。 Galectin-3被认为通过感光细胞降级在视网膜中起抗凋亡作用。（2）使用抗低星核蛋白抗体分离了与感光细胞（MLGAPC）相关的粘蛋白样糖蛋白（MLGAPC）的粘蛋白类糖蛋白的CDNA。不仅在photoreceptor基质中，而且在突触裂隙基质中检测到MLGAPC。通过检查MLGAPC在发展大鼠视网膜中的表达，我们确定低分子类型和高分子类型分别对应于锥和杆类型。由于感光细胞成熟后，MLGAPC的mRNA表达降低，因此MLGAPC的周转率被认为是缓慢的。蛋白质的这一特征对于MLGAPC保持了光感受器和外突触层的稳定结构。（3）将MLGAPC的人核蛋白的cDNA引入Y79视网膜细胞中。凝集素印迹分析表明，它们用末端唾液酸产生了杆型MLGAPC。由于在将透明质酸添加到培养基中后未观察到培养细胞的形态变化，因此MLGAPC和透明质酸之间的相互作用被认为不足以用于细胞粘附。