β-Catenin gain of function mutant in mouse periocular neural crest-derived mesenchymal cells impairs embryonic eyelid morphogenesis and leads to blepharophimosis syndrome in mice

Abstract

Purpose

The aberrant canonical Wnt-β-catenin signaling can cause devastating outcomes of tissue morphogenesis and tumor formation. In this study, we examined the impact of overexpression of constitutive active β-catenin in mouse periocular neural crest-derived mesenchymal cells during embryonic eyelid morphogenesis.

Methods

We expressed a stabilized β-catenin in which the exon 3 of the Ctnnb1 gene was deleted in periocular neural crest (PONC)-derived eyelid stromal cells (Ctnnb1^Δex3-PONC). Histopathological examinations were performed to examine the eyelid morphogenetic alterations in Ctnnb1^Δex3-PONC mice. Immunohistochemical investigations for cell proliferation, apoptosis, and differentiation were also assessed.

Results

We discovered that nuclear accumulation of β-catenin resulted in a reduction of nuclear Ki-67 and phospho-Erk1/2 expression levels and elevation of apoptosis in PONC cells during embryonic eyelid closure morphogenesis. Interestingly, however, the eyelid epithelial migration was not affected, which resulted in only eyelid epidermal closure but lacked underneath dermal formation at embryonic (E) day 16.5. The sequelae of Ctnnb1^Δex3-PONC revealed the malformation of the eyelid margin and Meibomian gland and deficiency of Muller's smooth muscle fibers formation. Consequently, Ctnnb1^Δex3-PONC mice manifested blepharophimosis syndrome at P21.

Conclusion

Our data suggested that aberrant expression of β-catenin gain of function in PONC interrupts the interplay between epithelium and stroma for the morphogenesis of eyelid closure during embryonic development.