EPITHELIAL SHEET MOVEMENT - EFFECTS OF TUNICAMYCIN ON MIGRATION AND GLYCOPROTEIN-SYNTHESIS
EPITHELIAL SHEET MOVEMENT - EFFECTS OF TUNICAMYCIN ON MIGRATION AND GLYCOPROTEIN-SYNTHESIS
复制标题
DOI：
10.1016/0012-1606(84)90131-3
复制
发表时间：
1984-01-01
影响因子：
2.7
通讯作者：
BRENNAN, SJ
中科院分区：
文献类型：
--
作者：
GIPSON, IK;KIORPES, TC;BRENNAN, SJ
在衣霉素（TM）（1 μ g/ml）（一种抑制天冬酰胺连接的糖蛋白的糖基化的抗生素）存在下，对直径为3 mm的具有中央上皮伤口的角膜进行器官培养。与在22小时内愈合的对照角膜相比，在TM存在下培养整个培养时间或仅前6小时的角膜显示出逐渐变慢的上皮愈合速率，其在培养时间的24小时基本上下降到零。在24小时，75%的伤口被覆盖。用无TM培养基反复洗涤（6次，10分钟每），这种效果可以始终被逆转角膜暴露于TM为6 hr. Incorporation的[3 H]葡糖胺到三氯乙酸沉淀蛋白的迁移上皮片减少到14%，对照组培养12小时后与TM，而[14 C]亮氨酸掺入没有显着影响。减少糖基化反映在细胞表面上培养12和20小时后，在TM的存在下：顶端细胞膜的前6个细胞的前缘的迁移片绑定显着较少的铁蛋白-伴刀豆球蛋白A颗粒每微米的膜比对照组。显然，天冬酰胺连接的糖蛋白的合成是角膜上皮层持续迁移所必需的。迁移所需的天冬酰胺连接的糖蛋白可能包括细胞表面糖蛋白。
Corneas with central epithelial wounds, 3 mm in diameter, were organ cultured in the presence of tunicamycin (TM) (1 .mu.g/ml), an antibiotic that inhibits glycosylation of asparagine-linked glycoproteins. Compared with control corneas, which healed in 22 h, corneas cultured in the presence of TM for the entire culture time or for only the first 6 h displayed a progressively slower epithelial healing rate that essentially dropped to zero by 24 h of culture time. At 24 h, .apprx. 75% of the wound was covered. After repeated washings with TM-free culture media (6.times., 10 min each), this effect could consistently be reversed in corneas exposed to TM for 6 hr. Incorporation of [3H]glucosamine into trichloroacetic acid-precipitable proteins of migrating epithelial sheets was reduced to 14% that of controls after 12 h of culture with TM, whereas [14C]leucine incorporation was not significantly affected. The decreased glycosylation was reflected on the cell surface after 12 and 20 h culture in the presence of TM: apical cell membranes of the first 6 cells of the leading edge of the migrating sheet bound significantly fewer ferritin-concanavalin A particles per micrometer of membrane than did controls. Apparently, synthesis of asparagine-linked glycoproteins is required for continued migration of corneal epithelial sheets. The asparagine-linked glycoproteins that are required for migration probably include cell-surface glycoproteins.