Response of aggregating chick corneal cells to modifiers of N-linked oligosaccharides, endoglycosidase H and deoxymannojirimycin.
Chick corneal epithelium takes on its mature conformation between 11 and 16 days of incubation. Earlier work has shown that desmosome frequency increases during this period, reaching its highest rate at 15 1/2 days. In the present report aggregation rates of cells from embryos of 11 days and those of 15 1/2 days are compared. Younger cells, which form fewer desmosomes, aggregate at a more moderate rate than older cells. In addition, younger cells bind less concanavalin A (ConA) than older cells. To determine if increase in ConA binding could be related to these cellular responses, aggregating cells were exposed to endoglycosidase H (EndoH) and to deoxymannojirimycin. This treatment should permit comparison of the response of cells that have a normal complement of N-linked oligosaccharides with those that have reduced high-mannose or complex type sugars. The effectiveness of EndoH under the conditions used was confirmed by failure of treated glycoprotein after separation by SDS-PAGE and electroblotting to bind ConA. Aggregation rates of both older and younger cells were unaffected, as measured by disapperance of single cells, though older cells formed somewhat smaller aggregates at the highest dosage used. Desmosome formation was markedly reduced in the presence of the enzyme, even in the absence of other changes in the fine structure. At the highest dose of the enzyme the fine structure of older but not younger cells showed indications of blockage of transport. Deoxymannojirimycin appears to cause a build-up of high-mannose groups, since treated cells showed increased incorporation of [3H]mannose.(ABSTRACT TRUNCATED AT 250 WORDS)
Overton
J
J
Department of Molecular Genetics & Cell Biology, University of Chicago, IL 60637.
D017485:1-Deoxynojirimycin; D000118:Acetylglucosaminidase / Q000494:pharmacology*; D000818:Animals; D002449:Cell Aggregation / Q000187:drug effects; D002478:Cells, Cultured; D002642:Chick Embryo; D003208:Concanavalin A / Q000378:metabolism; D003315:Cornea / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D003896:Desmosomes / Q000378:metabolism; D005944:Glucosamine / Q000031:analogs & derivatives / Q000494:pharmacology; D006596:Hexosaminidases / Q000494:pharmacology*; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D008854:Microscopy, Electron
D003208:Concanavalin A; D017485:1-Deoxynojirimycin; D006596:Hexosaminidases; D000118:Acetylglucosaminidase; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D005944:Glucosamine