Immunohistochemical localization of delta opioid receptors in peripheral tissues

The distribution of delta opioid receptor (DOR) immunoreactivity (ir) was examined in various peripheral tissues of Sprague-Dawley rats and macaque monkeys, including glabrous and hairy skin, corneas, eyelids, and the lip. DOR-ir was observed in all tissues examined. In addition to the presence of DOR-immunoreactive fibers in subcutaneous nerve bundles and the papillary dermis, we report the existence of positively labeled fibers and terminals in close association with peripheral structures not traditionally assigned a primarily nociceptive function, such as hair follicles, glandular apparatus, and blood vessels. In every case, staining was restricted to small-diameter axons that appeared to terminate as free nerve endings. To further classify DOR-immunoreactive fibers, we examined the extent of colocalization between DOR and three commonly used neuronal subtype markers; tyrosine hydroxylase (TH), calcitonin gene-related peptide (CGRP), and RT-97, a monoclonal antibody which preferentially labels neurons with myelinated axons. Additional double-labeling experiments using the nonspecific neuronal marker Protein Gene Product 9.5 were performed in glabrous skin to determine the percentage of total fiber count that displayed DOR-ir. No colocalization was observed between DOR and RT-97, indicating that DOR-ir is localized to unmyelinated axons. In addition, DOR colocalized with CGRP, but did not colocalize with TH. Taken together, these data support the hypothesis that delta opioid receptors in peripheral tissues are associated with sensory fibers, but not with the terminals of postganglionic sympathetic neurons.