The distribution and chemical coding of neurons in the porcine left and right inferior mesenteric ganglion projecting to the ascending colon and rectum have been investigated by using combined retrograde tracing and double-labelling immunohistochemistry. The ganglion contained many neurons supplying both gut regions. The colon-projecting neurons (CPN) occurred exclusively in the cranial part of the ganglia where they formed a large cluster distributed along the dorso-lateral ganglionic border and a smaller cluster located close to the caudal colonic nerve output. The rectum-projecting neurons (RPN) formed a long stripe along the entire length of the lateral ganglionic border and, within the right ganglion only, a small cluster located close to the caudal colonic nerve output. Immunohistochemistry revealed that the vast majority of the CPN and RPN were noradrenergic (tyrosine-hydroxylase-positive). Many noradrenergic neurons supplying the colon contained somatostatin or, less frequently, neuropeptide Y. In contrast, a significant subpopulation of the noradrenergic RPN expressed neuropeptide Y, whereas only a small proportion contained somatostatin. A small number of the non-adrenergic RPN were cholinergic (choline-acetyltransferase-positive) and a much larger subpopulation of the nerve cells supplying both the colon and rectum were non-adrenergic and non-cholinergic. Many cholinergic neurons contained neuropeptide Y. The non-adrenergic non-cholinergic neurons expressed mostly somatostatin or neuropeptide Y and some of those projecting to the rectum contained nitric oxide synthase, galanin or vasoactive intestinal polypeptide. Many of both the CPN and RPN were supplied with varicose nerve fibres exhibiting immunoreactivity against Leu5-enkephalin, somatostatin, choline-acetyltransferase, vasoactive intestinal polypeptide or nitric oxide synthase The somatotopic and neurochemical organization of this relatively large population of differently coded inferior mesenteric ganglion neurons projecting to the large bowel indicates that these cells are probably involved in intestino-intestinal reflexes controlling peristaltic and secretory activities.
The present study investigated: (1) the distribution and chemical coding of primary sensory neurons supplying the vas deferens in juvenile pigs by the use of retrograde tracing combined with double-labelling immunofluorescence, (2) nerve pathways from dorsal root ganglia (DRG) to the vas deferens by means of denervation procedures involving transection of the hypogastric or pelvic nerve combined with a retrograde tracing method, and (3) possible interactions of the substance P (SP)/calcitonin gene-related peptide (CGRP)-immunoreactive varicose nerve fibres on vas deferens projecting neurons (VDPN) in the anterior pelvic ganglion (APG). The vast majority of VDPN were found mainly in the lumbar L2, L3 and sacral S2, S3 pairs of DRG and showed a clear ipsilaterally organized projection pattern. Immunohistochemistry revealed that most of these neurons contained SP and/or CGRP, occasionally coexpressed with galanin. Interestingly, pronounced differences in the expression of SP and/or CGRP were observed between the lumbar and sacral VDPN in that most of the lumbar but less than half of the sacral neurons stained for these peptides. Denervation experiments showed that the neurons located within the lumbar DRG project through the ipsilateral hypogastric nerve, whereas those found within the sacral DRG send their processes through the ipsilateral and contralateral pelvic nerve. In the nerve-lesioned animals, especially in those with the hypogastric nerve cut, a dramatic reduction in the number of SP and/or CGRP-containing nerve terminals surrounding the efferent VDPN within the APG was observed. This study has disclosed the distribution and, for the first time, chemical coding and nerve pathways of vas deferens-projecting primary sensory neurons in a mammalian species, the pig. The results obtained also provide some novel information about the possible morphological and functional relationship between vas deferens-projecting primary sensory and pelvic efferent nerve cells.
The distribution and chemical coding of neurons supplying urinary bladder in the male pig were studied in the sympathetic chain ganglia, inferior mesenteric ganglia and anterior pelvic ganglia. The combined retrograde tracing and immunohistochemistry for tyrosine hydroxylase (TH), dopamine beta-hydroxylase (DBH), neuropeptide Y (NPY), somatostatin(SOM), galanin (GAL), vasoactive intestinal polypeptide(VIP), nitric oxide synthase (NOS), calcitonin gene related peptide (CGRP), substance P (SP), choline acetyltransferase (ChAT) and vesicular acetylcholine transporter(VAChT) were applied in the experiment. Bladder projecting neurons were found in all the ganglia studied. The majority of sympathetic ganglia neurons (inferior mesenteric ganglia and sympathetic chain ganglia) expressed immunoreactivity(IR) to DBH. In sympathetic chain ganglia these neurons simultaneously expressed NPY, GAL or VAChT,while in inferior mesenteric ganglia they contained NPY, SOM and/or GAL. A small number of these bladder projecting neurons was VAChT-IR and some contained NPY. In the pelvic ganglia bladder-projecting neurons formed two populations: DBH- and VAChT-IR. Some of DBH-IR neurons contained IR to NPY, SOM or GAL, while VAChTIR neurons were NPY-, SOM- or NOS-IR. The results indicate that sympathetic ganglia contain mainly adrenergic neurons,while pelvic ganglia contain both adrenergic and cholinergic neurons. All these neurons contain typical combinations of neuropeptides.
The present results suggest that inflammatory processes of the porcine ileum are able to induce changes in the intraganglionic architecture of a sympathetic ganglion located at discrete distance from the affected bowel segment.
The expression of galanin (GAL) and its three receptors (GalR1, GalR2, and GalR3) were studied with real-time PCR in the colonic wall of pigs suffering from experimental colitis caused by the infection with Brachyspira hyodysenteriae. The expression was studied in the muscular membrane, mucosa/submucosa layer, and in lymphocytes isolated from mucosa/submucosa. The expression levels were normalized to glyceraldehyde-6-phosphate dehydrogenase (GAPDH) expression and compared to expression levels in control animals. GAL expression was found in all three studied compartments of the colonic wall. A significant decrease in GAL expression level was found in the mucosa/submucosa and in isolated lymphocytes, whereas the decrease was much less profound in the muscular membrane. In the case of galanin receptors their expression was found in all studied compartments of the colonic wall, however at different levels, as compared to GAPDH expression. The decrease of galanin receptors expression was found in all studied compartments of the colonic wall of the sick animals.
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