The distribution of proenkephalin-derived peptides in the central nervous system of turtles

Abstract

The present study was carried out to examine if peptides similar to the various opioid peptide products of mammalian proenkephalin are present in the turtle central nervous system and to determine their distribution. Antisera against several enkephalin peptides were used: (1) leucine-enkephalin (LENK), (2) methionine-enkephalin (MENK), (3) methionine-enkephalin-arg 6 -phe 7 (MERF), (4) methionine-enkephalin-arg 6 -gly 7 -leu 8 (MERGL), (5) Peptide E (PEPE), and (6) BAM22P. Their specificity and cross-reactivity were carefully examined. The results indicated that LENK, MENK, and MERF (or highly similar peptides) are present in the turtle central nervous system, and that a peptide showing immunological similarity to BAM22P and PEPE also appeared to be present. In contrast, MERGL did not appear to be present. The distributions of the immunoreactive labeling for LENK, MENK, MERF, BAM22P, and PEPE were indistinguishable, and double-label studies showed that LENK, MERF, and BAM22P were colocalized within individual neurons and fibers. Although all of the above substances were observed in the same cell groups, there was some regional variation, in terms of which enkephalin peptide appeared to be most abundant. The distributions of these enkephalin peptides were very similar to those previously described in mammals and birds. Enkephalin was more abundant in the basal ganglia than in overlying telencephalic regions. Within the basal ganglia, enkephalin was present in striatal neurons and fibers and in pallidal fibers, thereby suggesting the existence of an enkephalinergic striatopallidal projection. Sensory relay nuclei of the thalamus were generally poor in enkephalinergic fibers, whereas the hypothalamus was rich in enkephalinergic neurons and fibers. Enkephalinergic neurons and fibers were present in the midbrain central gray. As is true of neurons of the nucleus spiriformis lateralis of the avian pretectum, the neurons of the homologous cell group in turtles, the dorsal nucleus of the posterior commissure of the pretectum, were found to contain enkephalin and have an enkephalinergic projection to the deep layers of the ipsilateral tectum. Enkephalinergic neurons and fibers were also abundant in the entry zones of the trigeminal nerve and dorsal root fibers of the spinal cord. The present results indicate that: (1) consistent with previously published biochemical studies (Lindberg and White, '86), proenkephalin in reptiles is similar in structure to that of mammals and, with the exception of MERGL, gives rise to similar or identical enkephalin peptides, and (2) the enkephalin peptides are found in many of the same systems of reptilian brain as mammalian and avian brain, and, therefore, may play a role in similar functions (e.g., basal ganglia motor functions) as in mammals and birds.