Wheat germ agglutinin and concanavalin a binding during epithelial wound healing in the cornea
作者:
McLaughlinBarbara J.,
BarlarEmily K.,
DonaldsonDonald J.,
期刊:
Current Eye Research
(Taylor Available online 1986)
卷期:
Volume 5,
issue 8
页码: 601-609
ISSN:0271-3683
年代: 1986
DOI:10.3109/02713688609015125
出版商: Taylor&Francis
数据来源: Taylor
摘要:
It has been hypothesized that there are differences between membrane-associated gly-coconjugates of wounded (migrating) epithelium and those of nonwounded (stationary) epithelium. To test this hypothesis, wheat germ agglutinin-ferritin (WGA-Fe) and concanavalin A-ferritin (Con A-Fe) binding to apical membranes of wounded and nonwounded rabbit corneal epithelia were compared. Epithelial abrasions of the superior half of the cornea were allowed to heal in vivo/ for six hours. Fixed corneas were then Incubated with lectin-ferritin and prepared for electron microscopy. Measurements (ferritin particles per linear um of membrane) of WGA-Fe Indicated that binding to leading cells (40.7 particles/um), to areas 20 to 35 cells behind the leading edge (46.5 particles/um) and to nonwounded epithelium (45.1 particles/um) from contra lateral eyes were not significantly different. A competitive inhibitor of WGA, 0.2M N-acetylglucosamine, however, blocked 94 percent of WGA binding on leading cells (2.3 particles/um), while binding persisted In areas behind the leading edge (39.5 particles/um) and on nonwounded epithelium (43.6 particles/um). This indicates that leading cell surfaces have a weak affinity for WGA. Unlike WGA, Con A showed a distinct preference for leading-edge cells (33.9 particles/um) compared to nonwounded epithelium (9 particles/um). In areas 20-35 cells behind the leading edge. Con A binding was Intermediate to these two extremes. This differential in Con A binding to the cells In wounded epithelium compared to nonwounded epithelium suggests that wounding Induces a change In the composition of membrane-associated gly-coconjugates. Evidence that Con A was binding primarily to simple of igosaccharides In each test area was obtained when endoglycosidase H, an enzyme which hydrolyzes high mannose glycoconjugates at their di-N-acetylchitobiose core, reduced binding by at least 60% In each test area. Taken together these data support the contention that there are differences In lectin receptor sites on wounded and nonwounded corneal epithelia.
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