Ultrastructure of substance P immunoreactive elements in the periaqueductal gray matter of the rat

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Abstract
Substance P (SP) is a non‐opioid peptide that generates a potent, analgesia when injected into the periaqueductal gray matter (PAG)The aim of this study was to investigate the fine neuronal structures and synaptic circuits involved in SP action in rats by means of electron microscopy, using immunocytochemical (ICC) pre‐embedding methods. A conventional ultrastructural study, carried out to interpret the ICC data correctly, shows small sized nerve cell bodies with a high nucleus–cytoplasmic ratio; absence of an extensive granular endoplasmic reticulum; and few axo‐somatic contacts having symmetrical and asymmetrical junctions in equal proportions. The large neuropil is characterized by numerous thin unmyelinated axons and axodendritic synapses mainly showing pleomorphic vesicles and asymmetrical junctions. The ICC analysis showed moderately labeled nerve cell bodies with the same structural, synaptic, and dimensional features as the negative cells. In the neuropil SP immunoreactivity is shown by dendrites, synapses, and thin elements which are unidentifiable structurally. No SP terminals synapsing on SP nerve cell bodies were found and only occasional SP light labeled terminals synapsing on negative perikarya were seen. The SP boutons generally have pleomorphic vesicles and asymmetrical junctions. On the basis of these data a possible excitatory activity of PAG SP synapses could be hypothesized. This activity would take place on postsynaptic neurons generally at a dendritic level. Our ultrastructural findings give support to an excitatory role carried out by SP neurons of the PAG, as suggested by the role of PAG circuitry on spinal nociception.