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Disruption of eyelid and cornea morphogenesis by epithelial β-catenin gain-of-function.

Mizoguchi S, Suzuki K, Zhang J, Yamanaka O, Liu CY, Okada Y, Miyajima M, Kokado M, Kao WW, Yamada G, Saika S - Mol. Vis. (2015)

Bottom Line: The ultrastructure of the ocular tissues of the E18.5 embryos was also examined.The mutant stroma exhibited impaired keratocyte differentiation with accelerated cell proliferation and reduction in the accumulation of collagen type I.The mutant embryos also showed hyperproliferative nodules in the ocular surface epithelia with anomaly of cornea-type epithelial differentiation and the absence of the epithelial basement membrane.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, Wakayama Medical University School of Medicine, Wakayama, Japan.

ABSTRACT

Purpose: To examine the developmental pathobiology of the eyelid and the cornea caused by epithelial β-catenin gain-of-function (gof) during mouse embryogenesis.

Methods: Compound mutant mice (Ctnnb1(GOFOSE) , gof of β-catenin in the epidermis and the ocular surface epithelium) were generated by time-mating keratin 5-promoter-Cre recombinase (Krt5-Cre) and Ctnnb1(fE3/WT) (floxed exon 3 of Ctnnb1) mice. Eyes obtained from wild-type (WT) and mutant embryos at various gestation stages until E18.5 were examined with histology and immunohistochemistry. The ultrastructure of the ocular tissues of the E18.5 embryos was also examined.

Results: Expression of the gof-β-catenin mutant protein in the epidermis severely impaired eyelid morphogenesis at E15.5, E17.5, and E18.5. The mutant stroma exhibited impaired keratocyte differentiation with accelerated cell proliferation and reduction in the accumulation of collagen type I. The mutant embryos also showed hyperproliferative nodules in the ocular surface epithelia with anomaly of cornea-type epithelial differentiation and the absence of the epithelial basement membrane.

Conclusions: Expression of the gof-β-catenin mutant protein in basal epithelial cells disrupts eyelid and cornea morphogenesis during mouse embryonic development due to the perturbation of cell proliferation and differentiation of the epithelium and the neural crest-derived mesenchyme.

No MeSH data available.


Related in: MedlinePlus

Ultrastructural and immunohistochemical examinations of impaired eyelids in mutant mice. Hematoxylin and eosin histology shows a clear difference in the cellular architecture of the epidermis and the mesenchyme of the eyelid (anlage) between the wild-type (WT) embryo (A) and the mutant embryo (B) with gain-of-function (gof)-β-catenin at E18.5. Nodular structures (srar) are observed in the mutant epidermis, but not in the WT epidermis. Mesenchymal cells are more densely packed (asterisk) in the mutant eyelid anlage than in the WT eyelid. Ultrastructural observation showed loss of stratification or intraepidermal differentiation in the upper layers with relatively round basal-like cells (Epi) and reduction of the accumulation of the collagenous matrix in the dermal tissue (asterisks) in an E18.5 mutant embryo (D) compared with the WT tissue (C). Immunohistochemistry shows laminin is well condensed in the subepithelial basement membrane zone in the WT embryo (E), while immunoreactivity for laminin is disrupted (arrows) beneath the epithelial nodular structure in the mutant eyelid anlage (F). The accumulation of β-catenin is detected in the nodular structures of the epidermis of the mutant embryo at E18.5 (arrows, H), while faint staining for β-catenin was seen in the epidermis of the WT embryo (G). The eyelid epidermis in the E18.5 WT and mutant embryos is not labeled for vimentin (I, J). The population of bromo-deoxyuridine (BrdU)-labeled epidermal keratinocytes was similar in the eyelid epidermis of the WT (K) and mutant embryos (L) at E15.5. Bar, 5 μm (C, D); 20 μm (E, F); 50 μm (A, B, G–J); 100 μm (K, L).
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f3: Ultrastructural and immunohistochemical examinations of impaired eyelids in mutant mice. Hematoxylin and eosin histology shows a clear difference in the cellular architecture of the epidermis and the mesenchyme of the eyelid (anlage) between the wild-type (WT) embryo (A) and the mutant embryo (B) with gain-of-function (gof)-β-catenin at E18.5. Nodular structures (srar) are observed in the mutant epidermis, but not in the WT epidermis. Mesenchymal cells are more densely packed (asterisk) in the mutant eyelid anlage than in the WT eyelid. Ultrastructural observation showed loss of stratification or intraepidermal differentiation in the upper layers with relatively round basal-like cells (Epi) and reduction of the accumulation of the collagenous matrix in the dermal tissue (asterisks) in an E18.5 mutant embryo (D) compared with the WT tissue (C). Immunohistochemistry shows laminin is well condensed in the subepithelial basement membrane zone in the WT embryo (E), while immunoreactivity for laminin is disrupted (arrows) beneath the epithelial nodular structure in the mutant eyelid anlage (F). The accumulation of β-catenin is detected in the nodular structures of the epidermis of the mutant embryo at E18.5 (arrows, H), while faint staining for β-catenin was seen in the epidermis of the WT embryo (G). The eyelid epidermis in the E18.5 WT and mutant embryos is not labeled for vimentin (I, J). The population of bromo-deoxyuridine (BrdU)-labeled epidermal keratinocytes was similar in the eyelid epidermis of the WT (K) and mutant embryos (L) at E15.5. Bar, 5 μm (C, D); 20 μm (E, F); 50 μm (A, B, G–J); 100 μm (K, L).

Mentions: Characterization of tissue phenotype was then performed with ultrastructural observation and immunohistochemistry. Transmission electron microscopy showed abnormal epidermal cell differentiation (loss of stratification or cornification in the superficial epithelial layer) and reduction in the collagenous matrix in the dermis of an E18.5 mutant embryo (Figure 3D) compared with that of a WT littermate (Figure 3C).


Disruption of eyelid and cornea morphogenesis by epithelial β-catenin gain-of-function.

Mizoguchi S, Suzuki K, Zhang J, Yamanaka O, Liu CY, Okada Y, Miyajima M, Kokado M, Kao WW, Yamada G, Saika S - Mol. Vis. (2015)

Ultrastructural and immunohistochemical examinations of impaired eyelids in mutant mice. Hematoxylin and eosin histology shows a clear difference in the cellular architecture of the epidermis and the mesenchyme of the eyelid (anlage) between the wild-type (WT) embryo (A) and the mutant embryo (B) with gain-of-function (gof)-β-catenin at E18.5. Nodular structures (srar) are observed in the mutant epidermis, but not in the WT epidermis. Mesenchymal cells are more densely packed (asterisk) in the mutant eyelid anlage than in the WT eyelid. Ultrastructural observation showed loss of stratification or intraepidermal differentiation in the upper layers with relatively round basal-like cells (Epi) and reduction of the accumulation of the collagenous matrix in the dermal tissue (asterisks) in an E18.5 mutant embryo (D) compared with the WT tissue (C). Immunohistochemistry shows laminin is well condensed in the subepithelial basement membrane zone in the WT embryo (E), while immunoreactivity for laminin is disrupted (arrows) beneath the epithelial nodular structure in the mutant eyelid anlage (F). The accumulation of β-catenin is detected in the nodular structures of the epidermis of the mutant embryo at E18.5 (arrows, H), while faint staining for β-catenin was seen in the epidermis of the WT embryo (G). The eyelid epidermis in the E18.5 WT and mutant embryos is not labeled for vimentin (I, J). The population of bromo-deoxyuridine (BrdU)-labeled epidermal keratinocytes was similar in the eyelid epidermis of the WT (K) and mutant embryos (L) at E15.5. Bar, 5 μm (C, D); 20 μm (E, F); 50 μm (A, B, G–J); 100 μm (K, L).
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Related In: Results  -  Collection

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f3: Ultrastructural and immunohistochemical examinations of impaired eyelids in mutant mice. Hematoxylin and eosin histology shows a clear difference in the cellular architecture of the epidermis and the mesenchyme of the eyelid (anlage) between the wild-type (WT) embryo (A) and the mutant embryo (B) with gain-of-function (gof)-β-catenin at E18.5. Nodular structures (srar) are observed in the mutant epidermis, but not in the WT epidermis. Mesenchymal cells are more densely packed (asterisk) in the mutant eyelid anlage than in the WT eyelid. Ultrastructural observation showed loss of stratification or intraepidermal differentiation in the upper layers with relatively round basal-like cells (Epi) and reduction of the accumulation of the collagenous matrix in the dermal tissue (asterisks) in an E18.5 mutant embryo (D) compared with the WT tissue (C). Immunohistochemistry shows laminin is well condensed in the subepithelial basement membrane zone in the WT embryo (E), while immunoreactivity for laminin is disrupted (arrows) beneath the epithelial nodular structure in the mutant eyelid anlage (F). The accumulation of β-catenin is detected in the nodular structures of the epidermis of the mutant embryo at E18.5 (arrows, H), while faint staining for β-catenin was seen in the epidermis of the WT embryo (G). The eyelid epidermis in the E18.5 WT and mutant embryos is not labeled for vimentin (I, J). The population of bromo-deoxyuridine (BrdU)-labeled epidermal keratinocytes was similar in the eyelid epidermis of the WT (K) and mutant embryos (L) at E15.5. Bar, 5 μm (C, D); 20 μm (E, F); 50 μm (A, B, G–J); 100 μm (K, L).
Mentions: Characterization of tissue phenotype was then performed with ultrastructural observation and immunohistochemistry. Transmission electron microscopy showed abnormal epidermal cell differentiation (loss of stratification or cornification in the superficial epithelial layer) and reduction in the collagenous matrix in the dermis of an E18.5 mutant embryo (Figure 3D) compared with that of a WT littermate (Figure 3C).

Bottom Line: The ultrastructure of the ocular tissues of the E18.5 embryos was also examined.The mutant stroma exhibited impaired keratocyte differentiation with accelerated cell proliferation and reduction in the accumulation of collagen type I.The mutant embryos also showed hyperproliferative nodules in the ocular surface epithelia with anomaly of cornea-type epithelial differentiation and the absence of the epithelial basement membrane.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, Wakayama Medical University School of Medicine, Wakayama, Japan.

ABSTRACT

Purpose: To examine the developmental pathobiology of the eyelid and the cornea caused by epithelial β-catenin gain-of-function (gof) during mouse embryogenesis.

Methods: Compound mutant mice (Ctnnb1(GOFOSE) , gof of β-catenin in the epidermis and the ocular surface epithelium) were generated by time-mating keratin 5-promoter-Cre recombinase (Krt5-Cre) and Ctnnb1(fE3/WT) (floxed exon 3 of Ctnnb1) mice. Eyes obtained from wild-type (WT) and mutant embryos at various gestation stages until E18.5 were examined with histology and immunohistochemistry. The ultrastructure of the ocular tissues of the E18.5 embryos was also examined.

Results: Expression of the gof-β-catenin mutant protein in the epidermis severely impaired eyelid morphogenesis at E15.5, E17.5, and E18.5. The mutant stroma exhibited impaired keratocyte differentiation with accelerated cell proliferation and reduction in the accumulation of collagen type I. The mutant embryos also showed hyperproliferative nodules in the ocular surface epithelia with anomaly of cornea-type epithelial differentiation and the absence of the epithelial basement membrane.

Conclusions: Expression of the gof-β-catenin mutant protein in basal epithelial cells disrupts eyelid and cornea morphogenesis during mouse embryonic development due to the perturbation of cell proliferation and differentiation of the epithelium and the neural crest-derived mesenchyme.

No MeSH data available.


Related in: MedlinePlus