Characterization of Porcine Ocular Surface Epithelial Microenvironment
The porcine ocular surface is used as a model of the human ocular surface; however, a detailed characterization of the porcine ocular surface has not been documented. This is due, in part, to the scarcity of antibodies produced specifically against the porcine ocular surface cell types or structures...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-04-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/8/7543 |
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| author | Naresh Polisetti Gottfried Martin Heidi R. Cristina Schmitz Ursula Schlötzer-Schrehardt Günther Schlunck Thomas Reinhard |
| author_facet | Naresh Polisetti Gottfried Martin Heidi R. Cristina Schmitz Ursula Schlötzer-Schrehardt Günther Schlunck Thomas Reinhard |
| author_sort | Naresh Polisetti |
| collection | DOAJ |
| description | The porcine ocular surface is used as a model of the human ocular surface; however, a detailed characterization of the porcine ocular surface has not been documented. This is due, in part, to the scarcity of antibodies produced specifically against the porcine ocular surface cell types or structures. We performed a histological and immunohistochemical investigation on frozen and formalin-fixed, paraffin-embedded ocular surface tissue from domestic pigs using a panel of 41 different antibodies related to epithelial progenitor/differentiation phenotypes, extracellular matrix and associated molecules, and various niche cell types. Our observations suggested that the Bowman’s layer is not evident in the cornea; the deep invaginations of the limbal epithelium in the limbal zone are analogous to the limbal interpalisade crypts of human limbal tissue; and the presence of goblet cells in the bulbar conjunctiva. Immunohistochemistry analysis revealed that the epithelial progenitor markers cytokeratin (CK)15, CK14, p63α, and P-cadherin were expressed in both the limbal and conjunctival basal epithelium, whereas the basal cells of the limbal and conjunctival epithelium did not stain for CK3, CK12, E-cadherin, and CK13. Antibodies detecting marker proteins related to the extracellular matrix (collagen IV, Tenascin-C), cell–matrix adhesion (β-dystroglycan, integrin α3 and α6), mesenchymal cells (vimentin, CD90, CD44), neurons (neurofilament), immune cells (HLA-ABC; HLA-DR, CD1, CD4, CD14), vasculature (von Willebrand factor), and melanocytes (SRY-homeobox-10, human melanoma black-45, Tyrosinase) on the normal human ocular surface demonstrated similar immunoreactivity on the normal porcine ocular surface. Only a few antibodies (directed against N-cadherin, fibronectin, agrin, laminin α3 and α5, melan-A) appeared unreactive on porcine tissues. Our findings characterize the main immunohistochemical properties of the porcine ocular surface and provide a morphological and immunohistochemical basis useful to research using porcine models. Furthermore, the analyzed porcine ocular structures are similar to those of humans, confirming the potential usefulness of pig eyes to study ocular surface physiology and pathophysiology. |
| first_indexed | 2024-03-11T04:55:07Z |
| format | Article |
| id | doaj.art-6348408b4a7d4cdaa41dec4fd52e87fa |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T04:55:07Z |
| publishDate | 2023-04-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-6348408b4a7d4cdaa41dec4fd52e87fa2023-11-17T19:42:02ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-04-01248754310.3390/ijms24087543Characterization of Porcine Ocular Surface Epithelial MicroenvironmentNaresh Polisetti0Gottfried Martin1Heidi R. Cristina Schmitz2Ursula Schlötzer-Schrehardt3Günther Schlunck4Thomas Reinhard5Eye Center, Medical Center, Faculty of Medicine, University of Freiburg, Killianstrasse 5, 79106 Freiburg, GermanyEye Center, Medical Center, Faculty of Medicine, University of Freiburg, Killianstrasse 5, 79106 Freiburg, GermanyCEMT-Freiburg, Experimental Surgery, Hospital—Medical Center, Faculty of Medicine, University of Freiburg, Breisacher Str. 66, 79106 Freiburg, GermanyDepartment of Ophthalmology, University Hospital Erlangen, Friedrich-Alexander-University of Erlangen-Nürnberg, Schwabachanlage 6, 91054 Erlangen, GermanyEye Center, Medical Center, Faculty of Medicine, University of Freiburg, Killianstrasse 5, 79106 Freiburg, GermanyEye Center, Medical Center, Faculty of Medicine, University of Freiburg, Killianstrasse 5, 79106 Freiburg, GermanyThe porcine ocular surface is used as a model of the human ocular surface; however, a detailed characterization of the porcine ocular surface has not been documented. This is due, in part, to the scarcity of antibodies produced specifically against the porcine ocular surface cell types or structures. We performed a histological and immunohistochemical investigation on frozen and formalin-fixed, paraffin-embedded ocular surface tissue from domestic pigs using a panel of 41 different antibodies related to epithelial progenitor/differentiation phenotypes, extracellular matrix and associated molecules, and various niche cell types. Our observations suggested that the Bowman’s layer is not evident in the cornea; the deep invaginations of the limbal epithelium in the limbal zone are analogous to the limbal interpalisade crypts of human limbal tissue; and the presence of goblet cells in the bulbar conjunctiva. Immunohistochemistry analysis revealed that the epithelial progenitor markers cytokeratin (CK)15, CK14, p63α, and P-cadherin were expressed in both the limbal and conjunctival basal epithelium, whereas the basal cells of the limbal and conjunctival epithelium did not stain for CK3, CK12, E-cadherin, and CK13. Antibodies detecting marker proteins related to the extracellular matrix (collagen IV, Tenascin-C), cell–matrix adhesion (β-dystroglycan, integrin α3 and α6), mesenchymal cells (vimentin, CD90, CD44), neurons (neurofilament), immune cells (HLA-ABC; HLA-DR, CD1, CD4, CD14), vasculature (von Willebrand factor), and melanocytes (SRY-homeobox-10, human melanoma black-45, Tyrosinase) on the normal human ocular surface demonstrated similar immunoreactivity on the normal porcine ocular surface. Only a few antibodies (directed against N-cadherin, fibronectin, agrin, laminin α3 and α5, melan-A) appeared unreactive on porcine tissues. Our findings characterize the main immunohistochemical properties of the porcine ocular surface and provide a morphological and immunohistochemical basis useful to research using porcine models. Furthermore, the analyzed porcine ocular structures are similar to those of humans, confirming the potential usefulness of pig eyes to study ocular surface physiology and pathophysiology.https://www.mdpi.com/1422-0067/24/8/7543limbal stem cellslimbal niche cellsmelanocyteslimbal epithelial progenitor cellsporcine ocular surfaceextracellular matrix |
| spellingShingle | Naresh Polisetti Gottfried Martin Heidi R. Cristina Schmitz Ursula Schlötzer-Schrehardt Günther Schlunck Thomas Reinhard Characterization of Porcine Ocular Surface Epithelial Microenvironment International Journal of Molecular Sciences limbal stem cells limbal niche cells melanocytes limbal epithelial progenitor cells porcine ocular surface extracellular matrix |
| title | Characterization of Porcine Ocular Surface Epithelial Microenvironment |
| title_full | Characterization of Porcine Ocular Surface Epithelial Microenvironment |
| title_fullStr | Characterization of Porcine Ocular Surface Epithelial Microenvironment |
| title_full_unstemmed | Characterization of Porcine Ocular Surface Epithelial Microenvironment |
| title_short | Characterization of Porcine Ocular Surface Epithelial Microenvironment |
| title_sort | characterization of porcine ocular surface epithelial microenvironment |
| topic | limbal stem cells limbal niche cells melanocytes limbal epithelial progenitor cells porcine ocular surface extracellular matrix |
| url | https://www.mdpi.com/1422-0067/24/8/7543 |
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