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Defective eyelid leading edge cell migration in C57BL/6-corneal opacity mice with an "eye open at birth" phenotype
Abstract:
Development of the eyelid requires coordination of the cellular processes involved in proliferation, cell size alteration, migration, and cell death. C57BL/6J-corneal opacity (B6-Co) mice are mutant mice generated by the administration of N-ethyl-N-nitrosourea (100 mg/kg). They exhibit the eyelids open at birth phenotype, abnormal round cell shape from tightened F-actin bundles in leading edge keratinocytes at E16.5, and gradual corneal opacity with neovessels. The tip of the leading edge in B6-Co mice did not move forward, and demonstrated a sharp peak shape without obvious directionality. Analysis of the biological characteristics of B6-Co mice demonstrated that abnormal migration of keratinocytes could affect eyelid development, but proliferation and apoptosis in B6-Co mice had no effect. Mutant gene mapping and sequence analysis demonstrated that in B6-Co mice, adenosine was inserted into the untranslated regions, between 3030 and 3031, in the mRNA 3'-terminal of Fgf10. In addition, guanine 7112 was substituted by adenine in the Mtap1B mRNA, and an A2333T mutation was identified in Mtap1B. Quantitative real-time polymerase chain reaction analysis showed that expression of the Hbegf gene was significantly down-regulated in the eyelids of B6- Co mice at E16.5, compared to B6 mice. However, the expression of Rock1, Map3k1, and Jnk1 genes did not show any significant changes. Abnormal keratinocyte migration and down-regulated expression of the Hbegf gene might be associated with impaired eyelid development in B6-Co mice.
Insights
Mutant mice with abnormal keratinocyte migration show impaired eyelid development, linked to down-regulated heparin-binding epidermal growth factor (Hbegf) gene expression. Proliferation and apoptosis were unaffected in this eyelid development study.
Area of Science:
- Developmental biology
- Genetics
- Cell biology
Background:
- Eyelid development is a complex process involving cellular proliferation, size changes, migration, and death.
- C57BL/6J-corneal opacity (B6-Co) mice, induced by N-ethyl-N-nitrosourea, display an 'eyelids open at birth' phenotype and corneal opacity.
Purpose of the Study:
- To investigate the cellular and genetic mechanisms underlying impaired eyelid development in B6-Co mice.
- To identify the specific cellular processes and genes affected in this mouse model.
Main Methods:
- Phenotypic analysis of B6-Co mice, including observation of eyelid opening and corneal opacity.
- Analysis of keratinocyte migration, proliferation, and apoptosis in mutant mice.
- Mutant gene mapping and sequence analysis to identify genetic alterations.
- Quantitative real-time polymerase chain reaction to assess gene expression levels.
Main Results:
- B6-Co mice exhibited abnormal keratinocyte shape and impaired leading edge migration, but normal proliferation and apoptosis rates.
- Genetic analysis revealed mutations in Fgf10 and Mtap1B mRNA in B6-Co mice.
- Significant down-regulation of heparin-binding epidermal growth factor (Hbegf) gene expression was observed in B6-Co mouse eyelids at E16.5.
- Expression of Rock1, Map3k1, and Jnk1 genes remained unchanged.
Conclusions:
- Abnormal keratinocyte migration is a key factor in the impaired eyelid development of B6-Co mice.
- Down-regulation of Hbegf gene expression may contribute to the observed developmental defects.
- The findings highlight the importance of keratinocyte migration and Hbegf in normal eyelid formation.
