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. 2015 Jan;185(1):197–213. doi: 10.1016/j.ajpath.2014.09.002

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© 2015 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.

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Eyelid closure defect and abnormal corneal development in Porcn^lox/Y;Wnt1-Cre;Rx3-Cre mutants. A: Control embryo at E17.5 with closed eyelid (arrow). B:Porcn^lox/Y;Wnt1-Cre;Rx3-Cre mutant at E17.5 with open eyelid, coloboma, and pigment abnormalities in the optic cup periphery. C: Frontal section of the chamber angle of control eye at E17.5 with closed eyelid (arrowhead) and developing ciliary body and iris (asterisk). Hematoxylin and eosin stain. D:Porcn^lox/Y;Wnt1-Cre;Rx3-Cre anterior segment at E17.5 shows growth-arrested eyelid folds (arrowhead), abnormal development of the peripheral retinal pigment epithelium (RPE) in the absence of ciliary body and iris (asterisk), and ectopic cells in the chamber angle (arrow). E: LEF1 expression in palpebral epidermis, mesenchyme (arrow), and conjunctival epithelium in upper eyelid of control eye at E15.5. F: In Porcn^lox/Y;Wnt1-Cre;Rx3-Cre embryos, LEF1 expression in the palpebral epidermis and conjunctival epithelium of the upper eyelid is maintained, but is absent in the mesenchyme (arrow). G: Expression of LEF1 in the lower eyelid in control embryo. H: In the lower eyelid of Porcn^lox/Y;Wnt1-Cre;Rx3-Cre embryos, LEF1 expression in the palpebral epidermis is decreased (right arrow), along with decreased expression in the mesenchyme (left arrow). I and K: Detection of p-JUN in the periderm, periderm extension (arrow), and conjunctival epithelium of upper (I) and lower (K) eyelids in control embryo at E15.5. J and L: In Porcn^lox/Y;Wnt1-Cre;Rx3-Cre eyelids at E15.5, p-JUN appears to be normal (arrows). M and N: Histological cross section of the cornea at E17.5 in control (M) and Porcn^lox/Y;Wnt1-Cre;Rx3-Cre eyes (N). Corneal thickness is indicated by a bracket; the corneal endothelium is marked by an arrow. O–X: Embryos at E15.5. O: PAX6 (red) expression in the corneal epithelium of control eyes (arrow). Nuclei are counterstained with DAPI (blue). P: PAX6 is present in the corneal epithelium in Porcn^lox/Y;Wnt1-Cre;Rx3-Cre eyes (arrow). Q and R: Expression of cytokeratin-12 in the corneal epithelium of control (Q) and Porcn^lox/Y;Wnt1-Cre;Rx3-Cre (R) embryos shows no difference (arrows). S: In control eyes, LEF1 is expressed in the corneal mesenchyme (arrow) and epithelium (arrowhead). T: LEF1 is decreased in the Porcn^lox/Y;Wnt1-Cre;Rx3-Cre corneal mesenchyme, except for some cells between lens and cornea (arrow). Reduced LEF1 expression in the corneal epithelium is variable (arrowhead). U: Robust PITX2 expression in control corneal mesenchyme (arrow). V: In Porcn^lox/Y;Wnt1-Cre;Rx3-Cre eyes, PITX2 is present in the hypocellular corneal mesenchyme (arrow). W: The Rho GTPase CDC42 is robustly expressed in the corneal mesenchyme in control eyes (arrow). X: CDC42 expression is severely reduced in Porcn^lox/Y;Wnt1-Cre;Rx3-Cre corneal mesenchyme (arrow). Scale bar = 100 μm.