Localisation of WE-14 immunoreactivity in the developing mouse limbo-corneal nerve net

WE-14 is generated in subpopulations of chromogranin A immunopositive endocrine cells and neurons including those innervating the anterior uvea. This study investigated WE-14 in intact sclero-limbo-corneal tissue from embryonic (E17), neonatal (N0-N16), and adult mice using immunocytochemistry and confocal scanning laser microscopy. Weak WE-14 immunostaining was observed at birth in nerve fibre tracts entering the corneal mid-stroma from the limbo-scleral junction. Immunopositive fibre tracts were evident throughout the cornea at N3; by N5 the mid-stromal plexus had begun to generate fibre populations extending toward the developing corneal epithelium, and some varicose fibres terminated amongst the developing epithelium. Immunostaining was evident at N7 in the developing limbo-scleral nerve net and some fibres exhibited a close association with unidentified vascular elements. By N11 and in subsequent neonates, the cornea had developed a distinct stratified nerve net composed of thick mid-stromal and thinner upper stromal nerve fibre bundles; both possessed populations of varicose WE-14 immunopositive fibres. In the adult, a sub-epithelial network of varicose WE-14 immunopositive fibres were evident at the limbo-scleral junction. Some fibres exhibited a close association with unidentified vascular elements, while others extended into the upper peripheral corneal stroma. WE-14 was evident in leashes throughout the basal corneal epithelium and generated fibres ramifying between the stratified epithelium with some fibres terminating amongst the outermost corneal epithelia. This study has demonstrated that WE-14 was evident in the limbo-corneal nerve net at birth and that its detection parallels corneal development to adulthood, where WE-14 is evident in a subpopulation of nerve fibres.