The role of epithelial membrane-associated mucin 4 (Muc4) in ocular surface health and corneal wound healing

Sara A Adelman¹Erin A Hisey¹Sangwan Park¹Nayone Lantyer-Araujo^1,2Melinda Quan¹Kathryn Sandberg¹Sabina Kahn¹Meher Khan¹Courtney Dreyer¹Sara Thomasy¹Kermit Carraway¹Joshua Morgan³Brian Leonard^1,2*

• ¹University of California Davis, Davis, United States
• ²Michigan State University, East Lansing, United States
• ³University of California Riverside, Riverside, United States

The purpose of this study was to determine the role of MUC4, a corneal membrane-associated mucin, on ocular surface health and corneal wounding healing using a Muc4 knockout (KO) mouse model. Complete ophthalmic examinations were performed on wildtype (WT), Muc4 heterozygous (Het) and Muc4 knockout (KO) mice, including slit lamp biomicroscopy, phenol red thread test (PRTT), intraocular pressure (IOP), and fluorescein staining. The mice were also assessed using optical coherence tomography (OCT), an advanced imaging technique. Dynamic contact angle goniometry was performed on ex vivo globes of WT, Muc4 Het and Muc4 KO mice to calculate contact angle hysteresis as a novel measure of the adherence properties of the corneal epithelium. To determine the effect of Muc4 in corneal wound healing, a phototherapeutic keratectomy (PTK) was performed on the right eye. After PTK wounding, the corneas were fluorescein stained, imaged, and the wound size was quantified using ImageJ at 0-, 24-, 36-, and 48-hours post-wounding. There were no phenotypic differences identified between WT, Muc4 Het and Muc4 KO mice on clinical examination, diagnostic testing, advanced imaging, and histology. While there was no difference in mucin 1 (Muc1) mRNA expression between WT and KO mice, there was a compensatory upregulation of a previously unreported murine corneal mucin, Muc20 mRNA expression, in corneal epithelium of Muc4 KO mice. No differences were detected between WT and Muc4 KO mice using dynamic contact angle goniometry. The Muc4 KO mice had significantly slower healing rates at 24h and 36h post-wounding when compared with WT mice (P < 0.05) and all healed by 48h post-wounding. These results support further investigation into compensatory roles of glycoproteins on the ocular surface, namely Muc4 and Muc20, and the role of Muc4 in epithelial cell migration in corneal wound healing.