Co-existence of neuropeptides in sympathetic, cranial autonomic and sensory neurons innervating the iris of the guinea-pig

We have used double-labelling immunofluorescence to identify the peptide content of autonomic and sensory neurons innervating the iris of albino guinea-pigs. Four major classes of neurons were identified on the basis of their distributions, origins and immunohistochemical characteristics. A dense plexus of noradrenergic axons in the constrictor and dilator muscles of the iris originated from the superior cervical ganglion, and contained immunoreactivity (IR) to both neuropeptide Y (NPY) and dynorphin (DYN). The constrictor and dilator muscles were also supplied with a dense plexus of axons with IR to substance P (SP). These axons probably originated from SP-IR nerve cell bodies located along the ciliary nerves, and are almost certainly the same axons as those producing cholinergic pupilloconstriction. The iris was also innervated by unmyelinated, capsaicin-sensitive axons with IR to both SP and calcitonin gene-related peptide. Most of these axons also contained IR to DYN and some were also IR for cholecystokinin. These axons are almost certainly sensory. Axons containing IR to both NPY and vasoactive intestinal peptide (VIP) were common in the ciliary processes, and also formed a sparse plexus near the ciliary margin of the dilator muscle. Following surgical sympathetic denervation these axons showed IR for dopamine-β-hydroxylase; they seemed to originate from the sphenopalatine ganglion. These results demonstrate that there are well-defined patterns of coexistence of neuropeptides in the autonomic and sensory neurons supplying the iris of guinea-pigs. To understand the physiological roles of these peptides, it will be necessary to consider the possibility of complex interactions between them.