Visceral afferent stimulation-evoked changes in the release of peptides into the parabrachial nucleus in vivo

Abstract

Previous investigations have demonstrated that the peptides substance P (SP), calcitonin gene-related peptide (CGRP), cholecystokinin (CCK), neurotensin (NT) and somatostatin (SOM) significantly modulate the glutamate-mediated transmission of visceral information through the parabrachial nucleus (PBN) to the ventrobasal thalamus. In addition, we have shown that the staining intensity of SOM, CCK and NT in the PBN decreases significantly following 2 h of vagal stimulation as visualized using immunohistochemistry. As well, the staining intensity of both SP and CGRP in the PBN were shown to increase under similar conditions. The present investigation was done to determine whether the altered peptide staining intensity of these peptides observed following 2 h of vagal stimulation was the result of an altered peptide release from terminals within the PBN. Male Sprague-Dawley rats were anesthetized with sodium thiobutabarbitol and instrumented to record blood pressure and heart rate and for the stimulation of the cervical vagus nerve. A push–pull perfusion cannula was lowered into the region of the PBN for the continuous sampling of extracellular fluid. Radioenzymatic quantification of the perfusates for peptide content revealed that the extracellular fluid concentration of CGRP and SP increased significantly during the 2 h of vagal stimulation. When the vagal stimulation was terminated, the release of both CGRP and SP decreased significantly below prestimulated values for approximately 30 min before returning to prestimulated levels shortly thereafter. In contrast, there was a significant decrease in the release of CCK, SOM and NT into the PBN during the period of vagal stimulation. Extracellular perfusate levels of these peptides returned to normal upon termination of stimulation. These results demonstrate that terminal release of CGRP and SP is significantly increased and terminal release of CCK, SOM and NT is significantly decreased in the PBN during 2 h of vagal stimulation. These results are consistent with our previous finding that the immunohistochemical staining intensity of CGRP and SP is increased while that of CCK, SOM and NT is decreased following vagal stimulation.