March 1996
Volume 37, Issue 4
Free
Articles  |   March 1996
Gangliosides of migrating and nonmigrating corneal epithelium in organ and cell culture.
Author Affiliations
  • Z Yang
    New England Eye Center, Boston, MA 02111, USA.
  • Z Zhao
    New England Eye Center, Boston, MA 02111, USA.
  • N Panjwani
    New England Eye Center, Boston, MA 02111, USA.
Investigative Ophthalmology & Visual Science March 1996, Vol.37, 501-510. doi:
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      Z Yang, Z Zhao, N Panjwani; Gangliosides of migrating and nonmigrating corneal epithelium in organ and cell culture.. Invest. Ophthalmol. Vis. Sci. 1996;37(4):501-510.

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      © ARVO (1962-2015); The Authors (2016-present)

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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.

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