Localization of immunoreactive dynorphin in neurons cultured from spinal cord and dorsal root ganglia.

Antisera specific for dynorphin were used to study the cellular distribution of opioid peptides in spinal cord and dorsal root ganglion neurons in dissociated cell culture. Radioimmunoassay of 4-wk-old cultures yielded levels of dynorphin immunoreactivity similar to those in adult rodent spinal cord. Immunohistochemistry showed staining confined to the perinuclear region of neuronal cell bodies. In contrast, enkephalin immunoreactivity was found in extensive neurite fields as well as in neuronal perikarya. Opioid peptide immunoreactivity was observed in approximately equal to 5% of the spinal cord neurons either with dynorphin or enkephalin antiserum. No substantial increase in the number of reactive cells was observed when the two sera were applied simultaneously. These results suggest that the perinuclear region of opioid spinal cord neurons in culture contains peptide with an amino acid sequence similar to that of the midportion of dynorphin, whereas the neurites appear to contain smaller peptides related to NH2-terminal fragments of dynorphin. By using simple morphological criteria, spinal sensory neurons can be identified in these cell cultures and in cultures prepared from dorsal root ganglia without spinal cord. Approximately 1-2% of these ganglion cells showed intense immunostaining with an affinity-purified dynorphin antiserum. An additional few percent of the sensory neurons showed less intense opioid immunoreactivity. This result extends the observations of opioid peptides one step further along the pathway that processes sensory information.