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LIM Homeobox Domain 2 Is Required for Corneal Epithelial Homeostasis.

Sartaj R, Chee RI, Yang J, Wan P, Liu A, Guaiquil V, Fuchs E, Rosenblatt MI - Stem Cells (2016)

Bottom Line: Immunodetection on corneal sections were used to visualize conjunctivalization, a sign of limbal barrier failure.Cell based assays showed that Lhx2cKO derived corneal epithelial cells have a significantly lower capacity to form colonies over time and delayed wound-healing recovery when compared to wildtype cells.We conclude that Lhx2 is required for maintenance of the corneal epithelial cell compartment and the limbal barrier.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, University of Illinois at Chicago, Chicago, Illinois, USA.

No MeSH data available.


Related in: MedlinePlus

Lhx2cKO mice exhibiting spontaneous corneal phenotypes P10 littermates from K14Cre+/− and Lhx2fl/fl gave rise to K14Cre−/− Lhx2fl/fl or WT (with hair) and K14Cre+/− Lhx2fl/fl or Lhx2cKO (reduced hair) (A). Slit lamp images of adult Lhx2cKO corneas from 6 to 8 weeks old mice (B) exhibiting the phenotype of Lhx2cKO cornea as clear and devoid of vessels (a), vascularized (b), perforated (c), and opaque infiltrated with vessels (d). Black arrows point to vessels in the cornea, white arrow to the perforation and red arrow to neovessels in the opaque cornea. Quantitative reverse transcriptase polymerase chain reaction from freshly isolated WT and Lhx2‐derived corneal epithelial cells showed a significant low level of expression of genes involved in cell stemness, including Lhx2, P63, and ABCG2(C). Data are means ± SE (n = 10). *, p ≤ 0.05. Abbreviation: WT, wild type.
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stem2257-fig-0002: Lhx2cKO mice exhibiting spontaneous corneal phenotypes P10 littermates from K14Cre+/− and Lhx2fl/fl gave rise to K14Cre−/− Lhx2fl/fl or WT (with hair) and K14Cre+/− Lhx2fl/fl or Lhx2cKO (reduced hair) (A). Slit lamp images of adult Lhx2cKO corneas from 6 to 8 weeks old mice (B) exhibiting the phenotype of Lhx2cKO cornea as clear and devoid of vessels (a), vascularized (b), perforated (c), and opaque infiltrated with vessels (d). Black arrows point to vessels in the cornea, white arrow to the perforation and red arrow to neovessels in the opaque cornea. Quantitative reverse transcriptase polymerase chain reaction from freshly isolated WT and Lhx2‐derived corneal epithelial cells showed a significant low level of expression of genes involved in cell stemness, including Lhx2, P63, and ABCG2(C). Data are means ± SE (n = 10). *, p ≤ 0.05. Abbreviation: WT, wild type.

Mentions: We next generated Lhx2cKO mice to study how the deletion of Lhx2 influences corneal epithelium homeostasis. These mice were obtained by breeding K14Cre+/− and Lhx2fl/fl and gave rise to K14Cre−/− Lhx2fl/fl or WT and K14Cre+/− Lhx2fl/fl or Lhx2cKO. When comparing postnatal day 10 (P10) littermates, it was clearly visible that the Lhx2cKO mice have reduced body hair when compared with WT (Fig. 2A). When we analyzed the cornea, all WT mice were normal (data not shown) and Lhx2cKO corneas were normal up to 6 weeks and devoid of vessels or ocular surface disorders (Fig. 2Ba). Some adult Lhx2cKO corneas exhibited a spontaneous phenotype including neovascularization (Fig. 2Bb), perforation (Fig. 2Bc), and opacification (Fig. 2Bd). These phenotypic changes became more common and severe as the mice aged (Table 1). For instance, at 6 weeks of age, 8% of mice showed neovascularization, that increased to 17% at 9 weeks, and become more prominent as with age, resulting in 83% of animal presenting this phenotype at 26 weeks. Perforated corneas sometimes occurred independently of vascularization and sometimes together (recorded at 8 weeks). Opacified corneas were noticed around 8 weeks old and increased rapidly over the weeks likely due to the healing from those mice exhibiting perforations. At 13 weeks, 25% of mice showed signs of neovascularization, while greater numbers (33%) were perforated. By 26 weeks only 16% of mice were normal in appearance while the majority were neovascularized, perforated, or opacified.


LIM Homeobox Domain 2 Is Required for Corneal Epithelial Homeostasis.

Sartaj R, Chee RI, Yang J, Wan P, Liu A, Guaiquil V, Fuchs E, Rosenblatt MI - Stem Cells (2016)

Lhx2cKO mice exhibiting spontaneous corneal phenotypes P10 littermates from K14Cre+/− and Lhx2fl/fl gave rise to K14Cre−/− Lhx2fl/fl or WT (with hair) and K14Cre+/− Lhx2fl/fl or Lhx2cKO (reduced hair) (A). Slit lamp images of adult Lhx2cKO corneas from 6 to 8 weeks old mice (B) exhibiting the phenotype of Lhx2cKO cornea as clear and devoid of vessels (a), vascularized (b), perforated (c), and opaque infiltrated with vessels (d). Black arrows point to vessels in the cornea, white arrow to the perforation and red arrow to neovessels in the opaque cornea. Quantitative reverse transcriptase polymerase chain reaction from freshly isolated WT and Lhx2‐derived corneal epithelial cells showed a significant low level of expression of genes involved in cell stemness, including Lhx2, P63, and ABCG2(C). Data are means ± SE (n = 10). *, p ≤ 0.05. Abbreviation: WT, wild type.
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stem2257-fig-0002: Lhx2cKO mice exhibiting spontaneous corneal phenotypes P10 littermates from K14Cre+/− and Lhx2fl/fl gave rise to K14Cre−/− Lhx2fl/fl or WT (with hair) and K14Cre+/− Lhx2fl/fl or Lhx2cKO (reduced hair) (A). Slit lamp images of adult Lhx2cKO corneas from 6 to 8 weeks old mice (B) exhibiting the phenotype of Lhx2cKO cornea as clear and devoid of vessels (a), vascularized (b), perforated (c), and opaque infiltrated with vessels (d). Black arrows point to vessels in the cornea, white arrow to the perforation and red arrow to neovessels in the opaque cornea. Quantitative reverse transcriptase polymerase chain reaction from freshly isolated WT and Lhx2‐derived corneal epithelial cells showed a significant low level of expression of genes involved in cell stemness, including Lhx2, P63, and ABCG2(C). Data are means ± SE (n = 10). *, p ≤ 0.05. Abbreviation: WT, wild type.
Mentions: We next generated Lhx2cKO mice to study how the deletion of Lhx2 influences corneal epithelium homeostasis. These mice were obtained by breeding K14Cre+/− and Lhx2fl/fl and gave rise to K14Cre−/− Lhx2fl/fl or WT and K14Cre+/− Lhx2fl/fl or Lhx2cKO. When comparing postnatal day 10 (P10) littermates, it was clearly visible that the Lhx2cKO mice have reduced body hair when compared with WT (Fig. 2A). When we analyzed the cornea, all WT mice were normal (data not shown) and Lhx2cKO corneas were normal up to 6 weeks and devoid of vessels or ocular surface disorders (Fig. 2Ba). Some adult Lhx2cKO corneas exhibited a spontaneous phenotype including neovascularization (Fig. 2Bb), perforation (Fig. 2Bc), and opacification (Fig. 2Bd). These phenotypic changes became more common and severe as the mice aged (Table 1). For instance, at 6 weeks of age, 8% of mice showed neovascularization, that increased to 17% at 9 weeks, and become more prominent as with age, resulting in 83% of animal presenting this phenotype at 26 weeks. Perforated corneas sometimes occurred independently of vascularization and sometimes together (recorded at 8 weeks). Opacified corneas were noticed around 8 weeks old and increased rapidly over the weeks likely due to the healing from those mice exhibiting perforations. At 13 weeks, 25% of mice showed signs of neovascularization, while greater numbers (33%) were perforated. By 26 weeks only 16% of mice were normal in appearance while the majority were neovascularized, perforated, or opacified.

Bottom Line: Immunodetection on corneal sections were used to visualize conjunctivalization, a sign of limbal barrier failure.Cell based assays showed that Lhx2cKO derived corneal epithelial cells have a significantly lower capacity to form colonies over time and delayed wound-healing recovery when compared to wildtype cells.We conclude that Lhx2 is required for maintenance of the corneal epithelial cell compartment and the limbal barrier.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, University of Illinois at Chicago, Chicago, Illinois, USA.

No MeSH data available.


Related in: MedlinePlus