Immunohistochemical localization of leucine-enkephalin in the spinal cord of the cat: enkephalin-containing marginal neurons and pain modulation

Abstract

This study examined the spinal cord distribution of the endogenous opioid peptide leucine-enkephalin in the cat using immunohistochemical techniques. The distribution of nerve processes was studied in untreated cats; colchicine was administered to study the distribution and morphology of spinal enkephalin-containing perikarya. Enkephalin immunoreactive processes were greatest in laminae I and II (marginal layer and substantia gelatinosa) of the superficial dorsal horn. In many sections, the outer substantia gelatinosa (SG), lamina IIa, was discernibly less immunoreactive than I or IIb. Laminae III and IV were relatively devoid of staining. Laminae V and VII had moderate enkephalin-immunoreactivity, lamina VI somewhat less. Enkephalin immunoreactivity in lamina X, around the central canal, was very dense. Enkephalin-containing beaded varicosities coursed throughout the ventral horn. Although previous studies in the rat emphasized the enkephalin-somata of the SG, we found that in the cat the majority of superficial dorsal horn enkephalin-somata are in the marginal layer. These enkephalin-containing marginal cells were morphologically similar to a population of marginal neurons which project to the brainstem and/or the thalamus. Some light staining small SG neurons were also identified; many were located at the lamina I-II border. Considerably more cells were found ventral to the SG, in lamina III, and at the IV-V border. These latter cells had dendrites coursing dorsally, toward the SG. Numerous immunoreactive cells were found in lamina VIII, in a band across the intermediate gray. These cells fused medially with cells of lamina X. Enkephalin cells were also found in the sacral autonomic nucleus and encircling the central cervical nucleus, Clarke's column, and stilling's nucleus. Although surrounded by labeled cells, the latter regions were devoid of enkephalin-immunoreactive processes. Many of these spinal enkephalin neurons are morphologically similar to and distributed in regions known to contain projection neurons. Thus it is suggested that many spinal enkephalin neurons, generally thought to be local circuit neurons, project rostrally, to other spinal levels and perhaps to brainstem and/or thalamus.