Immunohistochemical localization of tyrosine hydroxylase in corneal nerves

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Abstract
The sympathetic innervation of the mammalian cornea is thought to play an important role in the regulation of epithelial ion transport, mitogenesis, and wound healing following corneal injuries. Anatomically, the three‐dimensional organization and relative density of corneal sympathetic innervation in many species remains inadequately described. In the present study, the sympathetic innervation of five different mammals (guinea pig, rat, mouse, hamster, and human) was studied in corneas sectioned parallel to the main axis of fiber orientation by labeling the fibers immunohistochemically with antiserum against tyrosine hydroxylase and an avidin‐biotin‐diaminobenzidine technique. The results showed that each species displayed a distinctive pattern and density of tyrosine hydroxylase immunoreactive (TH‐IR) corneal innervation that was unique to that species. The overall level of TH‐IR innervation was highest in the guinea pig, moderate in the human, hamster, and rat, and lowest in the mouse. In all species examined, TH‐IR nerves were most numerous in the corneoscleral limbus where they either formed intimate associations with blood vessels or coursed through the connective tissue matrix apparently unrelated to vascular elements. Other TH‐IR nerves entered the cornea proper in radially directed stromal nerve bundles to give rise to subepithelial plexuses of varying complexity. Occasional intraepithelial penetrations were observed in the guinea pig, human, and rat. Removal of the superior cervical ganglion resulted in the total loss of TH‐IR staining from guinea pig and hamster corneas and in the substantial but incomplete loss of TH‐IR staining from rat and mouse corneas, thus demonstrating their predominantly sympathetic origin. Combined sympathetic and sensory ocular denervation in the rat eliminated almost all corneal and limbal TH‐IR immunostaining, thus suggesting a minor TH‐IR sensory component in this species. In agreement with this conclusion, small numbers of TH‐IR sensory neurons and an abundance of TH‐IR fibers were observed in the trigeminal ganglia of the rat and guinea pig. Removal of the rat main ciliary ganglion resulted in the loss of additional TH‐IR fibers from the chamber angle and iris, thereby confirming a partial parasympathetic contribution to the rat iridial TH‐IR innervation. Following unilateral removal of the superior cervical ganglion in rats and guinea pigs, the contralateral cornea contained increased numbers of TH‐IR nerves, suggesting an upregulation of tyrosine hydroxylase (TH) expression in some contralateral axons. In summary, these data have shown pronounced interspecies differences in the organization and density of the mammalian corneal TH‐IR innervation that may reflect species‐specific variation in the level of sympathetic control over select aspects of corneal physiology.