Haddad A, Bennett G
Am J Anat. 1987 Mar;178(3):259-68. doi: 10.1002/aja.1001780307.
3H-fucose was injected into the vitreous body of the eye(s) of 250-gm rats, which were then killed by means of an intracardiac perfusion with glutaraldehyde after intervals of 10 min, 1 and 4 hr, and 1 and 7 days. The eyes were removed and further fixed, and pieces of retina were processed for light and electron microscope radioautography. Light microscope radioautography showed that the pigment epithelial cells actively incorporated 3H-fucose label. The intensity of reaction peaked at 4 hr after injection of the label and then slowly declined. Quantitative electron microscope radioautography revealed that, at 10 min after 3H-fucose injection, over 70% of the label was localized to the Golgi apparatus, indicating that fucose residues are added to newly synthesized glycoproteins principally at this site. With time the proportion of label associated with the Golgi apparatus decreased, but that assigned to the infolded basal plasma membrane, the apical microvilli, and various apical lysosomes increased. These results indicate that in retinal pigment epithelial cells newly synthesized glycoproteins continuously migrate from the Golgi apparatus to lysosomes and to various regions of the plasma membrane. In this case, the membrane glycoproteins may play specific roles in receptor functions of the basal plasma membrane or phagocytic activities at the apical surface. Very little label migrated to Bruch's membrane, indicating either a very slow turnover or a paucity of fucose-containing glycoproteins at this site.
将3H-岩藻糖注入250克大鼠的眼玻璃体中,然后在间隔10分钟、1小时、4小时以及1天和7天后,通过用戊二醛进行心内灌注将大鼠处死。取出眼睛并进一步固定,将视网膜切片进行光镜和电镜放射自显影处理。光镜放射自显影显示色素上皮细胞积极摄取3H-岩藻糖标记。注射标记后4小时反应强度达到峰值,然后缓慢下降。定量电镜放射自显影显示,在注射3H-岩藻糖后10分钟,超过70%的标记定位于高尔基体,这表明岩藻糖残基主要在该部位添加到新合成的糖蛋白上。随着时间的推移,与高尔基体相关的标记比例下降,但分配给内褶的基底质膜、顶端微绒毛和各种顶端溶酶体的标记比例增加。这些结果表明,在视网膜色素上皮细胞中,新合成的糖蛋白不断从高尔基体迁移到溶酶体和质膜的各个区域。在这种情况下,膜糖蛋白可能在基底质膜的受体功能或顶端表面的吞噬活动中发挥特定作用。很少有标记迁移到布鲁赫膜,这表明该部位要么周转非常缓慢,要么含岩藻糖的糖蛋白很少。