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Gangliosides of migrating and nonmigrating corneal epithelium in organ and cell culture.

Abstract：

PURPOSE:To identify major gangliosides - the sialated glycolipids - of corneal epithelium; to determine which specific gangliosides, if any, are synthesized in a higher amount or are downregulated during corneal epithelial cell migration; and to determine what role, if any, they play in the modulation of corneal epithelial cell proliferation. METHODS:[3H]-galactose-labeled and unlabeled glycolipids of migrating and nonmigrating rabbit corneal epithelium in cell and/or in organ culture were chromatographed on DEAE Sephadex to isolate gangliosides. The gangliosides eluted from the ion-exchange column were further characterized by thin-layer chromatography (TLC), glycosidase digestions, and TLC-immunostain analysis. A [3H]-thymidine incorporation assay was used to determine the effect of exogenous gangliosides on corneal epithelium cell proliferation. RESULTS:Upon TLC of the acidic fraction eluted from the DEAE column, only two radiolabeled glycolipids (GL1 and GL2), migrating as a doublet, were detected. Regardless of whether the epithelia were prepared by cell culture or organ culture, both GL1 and GL2 were present in a significantly higher amount in migrating compared to nonmigrating epithelia. Further characterization of GL1 and GL2 identified them as gangliosides known as GM3. TLC-immunostain analysis, as well as orcinol staining of thin-layer chromatograms of gangliosides of unlabeled cells, revealed that GM3 also accumulates in a higher amount in migrating compared to nonmigrating epithelial cell cultures. Exogenous addition of GM3, but not various other gangliosides, inhibited corneal epithelial cell proliferation in a dose-dependent manner. CONCLUSIONS:GM3 is the major ganglioside present in corneal epithelium, and its levels are elevated during corneal epithelial cell migration. It is suggested that the ganglioside plays a role in events that modulate corneal epithelial cell proliferation.

journal_name

Invest Ophthalmol Vis Sci

journal_title

Investigative ophthalmology & visual science

authors

Yang Z,Zhao Z,Panjwani N

subject

Has Abstract

pub_date

1996-03-01 00:00:00

pages

501-10

issue

eissn

0146-0404

issn

1552-5783

journal_volume

pub_type

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Gangliosides of migrating and nonmigrating corneal epithelium in organ and cell culture.