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Expression of the 55-kD/64-kD corneal keratins in ocular surface epithelium
M A Kurpakus1, E L Stock, J C Jones
1Department of Cell Biology and Anatomy, Northwestern University Medical School, Chicago, Ill 60611.
Insights
A 55-kD/64-kD keratin pair marks corneal-type differentiation. Antibodies targeting these keratins identify specific ocular epithelial cells potentially involved in corneal wound healing.
Area of Science:
- Cell Biology
- Ophthalmology
- Biochemistry
Background:
- Keratin pairing, defined by tissue coexpression, identifies specific cell differentiation.
- A 55-kD/64-kD keratin pair is recognized as a marker for corneal-type differentiation.
Purpose of the Study:
- To investigate the distribution of 55-kD and 64-kD keratins in ocular epithelia.
- To explore the potential role of identified epithelial cells in corneal reepithelialization.
Main Methods:
- Generation of rabbit antiserum against a 55-kD polypeptide.
- Immunofluorescence microscopy and immunoblotting analysis using specific antisera and monoclonal antibody AE5.
- Analysis of guinea pig and bovine corneal and limbal epithelium.
Main Results:
- The 55-kD antiserum labeled corneal and limbal epithelium, including a subpopulation of peripheral limbal cells.
- Monoclonal antibody AE5 recognized a 64-kD polypeptide in guinea pig cornea and a 66-kD polypeptide in bovine cornea.
- AE5 labeled suprabasal layers of corneal and limbal epithelium and peripheral limbal cells.
Conclusions:
- The 55-kD and 64-kD/66-kD keratins are specific markers for corneal and limbal epithelial cells.
- Peripheral limbal cells identified by these keratins may play a role in corneal reepithelialization after injury.
Abstract:
According to the concept of keratin pairing defined by tissue coexpression, a 55-kD/64-kD keratin pair is a marker of "corneal-type" differentiation. Intermediate filament (IF)-enriched preparations from guinea pig and bovine corneal epithelium were analyzed, and a rabbit antiserum was generated against a 55-kD polypeptide enriched in these preparations. This antiserum generated a typical IF-like pattern in cultured bovine corneal epithelial cells. Immunofluorescence microscopic analysis of frozen sections of guinea pig and bovine tissue revealed that the 55-kD antiserum labeled corneal and limbal epithelium. In addition, the antiserum stained a subpopulation of peripheral limbal cells that were distributed in both basal and suprabasal layers of the epithelium. The monoclonal antibody AE5 was used to investigate the distribution of the 64-kD polypeptide in guinea pig and bovine tissue. Immunoblotting analysis revealed that AE5 antibodies recognized a 64-kD polypeptide in guinea pig cornea, but recognized a 66-kD polypeptide in bovine cornea. Immunofluorescence microscopic analysis of guinea pig tissue revealed that AE5 antibodies labeled suprabasal layers of corneal epithelium, in suprabasal layers of limbal epithelium, and in groups of cells in the peripheral limbal epithelium. We discuss the possibility that the ocular epithelial cells recognized by either the 55-kD or the 64-kD antibodies in the peripheral limbus may play a role in the reepithelialization of the cornea after wounding.
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