Fluctuations in Corneal Endothelial LAP2 Expression Levels Correlate with Passage Dependent Declines in Their Cell Proliferative Activity
The corneal endothelium is the inner corneal mono-layered epithelium, fundamental for preserving corneal hydration and transparency. However, molecular mechanisms that regulate corneal endothelial cells (CEnCs), in particular regarding their proliferative capacity, have been only partially elucidate...
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MDPI AG
2022-05-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/10/5859 |
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| author | Eleonora Maurizi Alessia Merra Davide Schiroli Benedetta Ghezzi Claudio Macaluso Graziella Pellegrini |
| author_facet | Eleonora Maurizi Alessia Merra Davide Schiroli Benedetta Ghezzi Claudio Macaluso Graziella Pellegrini |
| author_sort | Eleonora Maurizi |
| collection | DOAJ |
| description | The corneal endothelium is the inner corneal mono-layered epithelium, fundamental for preserving corneal hydration and transparency. However, molecular mechanisms that regulate corneal endothelial cells (CEnCs), in particular regarding their proliferative capacity, have been only partially elucidated. CEnCs are quiescent in vivo and they easily undergo endothelial to mesenchymal transition (EnMT) in vitro. This study aims to analyze CEnCs behavior and expression in vitro, either in sub-confluent growing (S) or confluent (C) CEnCs cultures. Primary rabbit and human CEnCs were cultured and used for RT-PCR, immunofluorescence or western blot analysis. These methods allowed identifying a novel molecular marker, LAP2, that is upregulated in S while downregulated in C human or rabbit CEnCs. Those results were observed for several subsequent passages in culture and this, together with the correlation between ki67 and LAP2 expression, suggested LAP2 as a novel possible indicator for culture ageing. Finally, treatment with FGF and TGFβ in rCEnCs highlighted how LAP2 can vary as the cells regulate their proliferative state. In conclusion, we have identified a novel marker for CEnCs, LAP2, that regulates its expression depending on the cells sub/confluent state and that correlates with CEnCs proliferation. |
| first_indexed | 2024-03-10T03:42:56Z |
| format | Article |
| id | doaj.art-ddaf8d72049e4701b6591bbbeae2cb7e |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T03:42:56Z |
| publishDate | 2022-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-ddaf8d72049e4701b6591bbbeae2cb7e2023-11-23T11:29:40ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-05-012310585910.3390/ijms23105859Fluctuations in Corneal Endothelial LAP2 Expression Levels Correlate with Passage Dependent Declines in Their Cell Proliferative ActivityEleonora Maurizi0Alessia Merra1Davide Schiroli2Benedetta Ghezzi3Claudio Macaluso4Graziella Pellegrini5Centre for Regenerative Medicine, University of Modena and Reggio Emilia, 41125 Modena, ItalyHolostem Terapie Avanzate S.r.l., 41125 Modena, ItalyTransfusion Medicine Unit, Azienda USL-IRCCS, 42123 Reggio Emilia, ItalyDentistry Centre Lab, University of Parma, 43126 Parma, ItalyDentistry Centre Lab, University of Parma, 43126 Parma, ItalyCentre for Regenerative Medicine, University of Modena and Reggio Emilia, 41125 Modena, ItalyThe corneal endothelium is the inner corneal mono-layered epithelium, fundamental for preserving corneal hydration and transparency. However, molecular mechanisms that regulate corneal endothelial cells (CEnCs), in particular regarding their proliferative capacity, have been only partially elucidated. CEnCs are quiescent in vivo and they easily undergo endothelial to mesenchymal transition (EnMT) in vitro. This study aims to analyze CEnCs behavior and expression in vitro, either in sub-confluent growing (S) or confluent (C) CEnCs cultures. Primary rabbit and human CEnCs were cultured and used for RT-PCR, immunofluorescence or western blot analysis. These methods allowed identifying a novel molecular marker, LAP2, that is upregulated in S while downregulated in C human or rabbit CEnCs. Those results were observed for several subsequent passages in culture and this, together with the correlation between ki67 and LAP2 expression, suggested LAP2 as a novel possible indicator for culture ageing. Finally, treatment with FGF and TGFβ in rCEnCs highlighted how LAP2 can vary as the cells regulate their proliferative state. In conclusion, we have identified a novel marker for CEnCs, LAP2, that regulates its expression depending on the cells sub/confluent state and that correlates with CEnCs proliferation.https://www.mdpi.com/1422-0067/23/10/5859LAP2corneal endotheliumfluctuatingmarkerconfluentproliferation |
| spellingShingle | Eleonora Maurizi Alessia Merra Davide Schiroli Benedetta Ghezzi Claudio Macaluso Graziella Pellegrini Fluctuations in Corneal Endothelial LAP2 Expression Levels Correlate with Passage Dependent Declines in Their Cell Proliferative Activity International Journal of Molecular Sciences LAP2 corneal endothelium fluctuating marker confluent proliferation |
| title | Fluctuations in Corneal Endothelial LAP2 Expression Levels Correlate with Passage Dependent Declines in Their Cell Proliferative Activity |
| title_full | Fluctuations in Corneal Endothelial LAP2 Expression Levels Correlate with Passage Dependent Declines in Their Cell Proliferative Activity |
| title_fullStr | Fluctuations in Corneal Endothelial LAP2 Expression Levels Correlate with Passage Dependent Declines in Their Cell Proliferative Activity |
| title_full_unstemmed | Fluctuations in Corneal Endothelial LAP2 Expression Levels Correlate with Passage Dependent Declines in Their Cell Proliferative Activity |
| title_short | Fluctuations in Corneal Endothelial LAP2 Expression Levels Correlate with Passage Dependent Declines in Their Cell Proliferative Activity |
| title_sort | fluctuations in corneal endothelial lap2 expression levels correlate with passage dependent declines in their cell proliferative activity |
| topic | LAP2 corneal endothelium fluctuating marker confluent proliferation |
| url | https://www.mdpi.com/1422-0067/23/10/5859 |
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