Multiple forms of pancreatic polypeptide-related compounds in the lamprey CNS: partial characterization and immunohistochemical localization in the brain stem and spinal cord

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Abstract
Although neuropeptide Y (NPY) is established as a transmitter in many regions of the nervous system, the role of other peptides of the pancreatic polypeptide (PP) family in the CNS is obscure. This study provides evidence that PP-like peptides in the “primitive” CNS of a cyclostome are composed of different molecular forms, which are stored in separate neuronal populations with apparently different functions. PP-like material was detected in extracts of brain and spinal cord from Lampetra fluviatilis by radioimmunoassay (RIA) using an antiserum to the C-terminal hexapeptide of mammalian PP. The PP-immunoreactive material consisted of several molecular forms, as shown by its complex elution profile on high-performance liquid chromatography (HPLC). The cellular distribution of PP-like immunoreactivity was studied with indirect immunofluorescence histochemistry using antisera toward porcine peptide YY (PYY), porcine neuropeptide Y (NPY), and bovine (BPP), rat (RPP), and avian (APP) pancreatic polypeptide. Adjacent sections from the brain stem and spinal cord of L. fluviatilis and Ichthyomyzon unicuspis, incubated with the different antisera, displayed 2 main patterns of PP immunoreactivity. The PYY and RPP antisera labeled groups of neurons and fibers in the rhombencephalic and mesencephalic reticular formation. One of the PYY/RPP-ir cell groups, located in the anterior rhombencephalic reticular nucleus, had a projection to the dorsolateral spinal cord. Fibers of this reticulospinal system were in close apposition to dendrites of intracellularly stained spinal motoneurons and sensory relay interneurons, indicating that they may receive PPergic input. In contrast, antisera to NPY and APP labeled local neurons systems in the spinal dorsal horn, in the lateral parts of the brain stem, including the rhombencephalic alar plate, and in the retina. The BPP antiserum recognized the NPY/APP as well as the PYY/RPP immunoreactive neuron systems, further supporting that they both contain PP-like peptides.