Abstract / Summary
Abstract Sex and hormonal status are clinically relevant determinants of ocular-surface physiology, but whether adult androgen exposure shifts multiple components of the ocular surface in parallel remains poorly defined. We compared vehicle-treated female mice with testosterone-treated females and used intact males as a physiological reference rather than as a procedure-matched control. Modified-nucleoside profiling, tear proteomics, corneal and trigeminal ganglion RNA sequencing, corneal flat-mount nerve volumetry, von Frey aesthesiometry, phospho-histone H3 labelling and single-endpoint Krt14-CreERT; CAG-Cytbow lineage tracing were combined to examine the tear-nerve-epithelial system at baseline. Tear collection rate was unchanged, whereas total tear protein concentration and proteomic composition differed between vehicle-treated and testosterone-treated females and shifted toward the intact male reference; 1,052 of 3,125 identified tear proteins were modulated in at least one pairwise comparison. Testosterone-treated females also showed lower central βIII-tubulin-positive nerve-fibre volume and mechanical sensitivity and fewer mitotic epithelial cells than vehicle-treated females, while lineage-labelled epithelial foci showed a qualitatively concordant distribution. Corneal transcriptional responses were substantially broader than those in the trigeminal ganglion. These findings show that adult testosterone exposure is associated with coordinated plasticity across several baseline ocular-surface phenotypes. They establish endocrine status as an important biological variable for experimental design while leaving its consequences for injury and disease to future functional studies.