Mariusz Majewski

Distribution and immunohistochemical characteristics of neurons in the porcine caudal mesenteric ganglion projecting to the vas deferens and seminal vesicle

Combined retrograde tracing (using fluorescent tracer Fast Blue) and double-labelling immunofluorescence were used to study the distribution and immunohistochemical characteristics of neurons in the porcine caudal mesenteric ganglion projecting to the vas deferens and seminal vesicle. The distribution and immunohistochemical properties of neurons projecting to both organs were similar. As revealed by retrograde tracing, Fast Blue-positive neurons were located within the left and right ganglia, with a distinct predominance in the ipsilateral one. In the ipsilateral ganglion, the majority of the neurons were located caudally, along the dorso-lateral ganglionic border, suggesting a somatotopic organization of the ganglion. Immunohistochemistry revealed four populations of retrogradely labelled neurons (from the largest to the smaller one): tyrosine hydroxylase-positive/neuropeptide Y-negative (TH+/NPY-), TH+/NPY+, TH-/NPY-, TH-/NPY+. With respect to their surrounding nerve fibres, two subpopulations of the dye-labelled neurons could be distinguished. The small one consisted of solitary neurons receiving a strong calcitonin gene-related peptide- and Leu5-enkephalin-, and a less intense vasoactive intestinal peptide-immunoreactive innervation. The remaining neurons were poorly supplied by singular nerve fibres containing some of the investigated peptides. We conclude that the caudal mesenteric ganglion should be considered as a prominent source of adrenergic and/or NPY-positive innervation for the porcine male reproductive tract.

Distribution of neurons innervating the uterus of the pig

The distribution of neurons innervating the uterus of the pig was studied with the use of fluorescent retrograde tracer Fast Blue. Tracer injections were made into the uterine cervix, pericervical, middle and perioviductal part of the right uterine horn. After tracer injection into the uterine cervix tracer-positive neurons were found bilaterally in the inferior mesenteric ganglia, paracervical ganglia, paravertebral ganglia L1-S3 and dorsal root ganglia Th10-L4 and S2-S3. After tracer injection into the pericervical part of the right uterine horn the distribution of tracer-positive neurons resembled the one described earlier, except the tracer-positive neurons were absent in the left dorsal root ganglia. After tracer injection into the middle part of the right uterine horn tracer-positive neurons were found in the ipsilateral inferior mesenteric ganglion, bilaterally in the paracervical ganglia and in the ipsilateral paravertebral ganglia (L2-L6). Injection of the tracer into the perioviductal part of the right uterine horn revealed tracer-positive neurons bilaterally in the inferior mesenteric ganglia and paracervical ganglia and single cells in the ipsilateral paravertebral ganglia.

Immunohistochemical properties of nerve fibres supplying accessory male genital glands in the pig. A colocalisation study.

Abstract Immunohistochemical studies have been performed to investigate the occurrence and coexistence of two catecholamine-synthesising enzymes, tyrosine hydroxylase and dopamine-β-hydroxylase, and several neuropeptides, including neuropeptide Y, vasoactive intestinal polypeptide, Leu5-enkephalin, somatostatin, calcitonin gene-related peptide and substance P, in nerve fibres supplying porcine accessory genital glands, the seminal vesicles, prostate (body and the disseminated part) and bulbourethral glands. Three major populations of nerve fibres supplying non-vascular elements of the glands have been distinguished (from the largest to the smallest one): (1) noradrenergic fibres, the majority of which contain Leu5-enkephalin, neuropeptide Y or, to a lesser extent, somatostatin, (2) non-noradrenergic, putative cholinergic fibres containing vasoactive intestinal polypeptide, neuropeptide Y and/or somatostatin and, (3) non-noradrenergic, presumably sensory fibres, containing calcitonin gene-related peptide and substance P. Whilst the coexistence patterns within nerves supplying particular glands are similar, the density of innervation varies between the organs. The innervation of the seminal vesicles and prostatic body is more developed than that of the disseminated part of the prostate and bulbourethral glands. The majority of noradrenergic fibres related to blood vessels contain neuropeptide Y only, while the non-noradrenergic nerves contain mainly vasoactive intestinal polypeptide. The possible function and origin of particular nerve fibre populations are discussed.

The Distribution and Chemical Coding of Intramural Neurons Supplying the Porcine Stomach – the Study on Normal Pigs and on Animals Suffering from Swine Dysentery

The present study was designed to investigate the expression of biologically active substances by intramural neurons supplying the stomach in normal (control) pigs and in pigs suffering from dysentery. Eight juvenile female pigs were used. Both dysenteric (n = 4; inoculated with Brachyspira hyodysenteriae) and control (n = 4) animals were deeply anaesthetized, transcardially perfused with buffered paraformalehyde, and tissue samples comprising all layers of the wall of the ventricular fundus were collected. The cryostat sections were processed for double‐labelling immunofluorescence to study the distribution of the intramural nerve structures (visualized with antibodies against protein gene‐product 9.5) and their chemical coding using antibodies against vesicular acetylcholine (ACh) transporter (VAChT), nitric oxide synthase (NOS), galanin (GAL), vasoactive intestinal polypeptide (VIP), somatostatin (SOM), Leu5‐enkephalin (LENK), substance P (SP) and calcitonin gene‐related peptide (CGRP). In both inner and outer submucosal plexuses of the control pigs, the majority of neurons were SP (55% and 58%, respectively)‐ or VAChT (54%)‐positive. Many neurons stained also for CGRP (43 and 45%) or GAL (20% and 18%) and solitary perikarya were NOS‐, SOM‐ or VIP‐positive. The myenteric plexus neurons stained for NOS (20%), VAChT (15%), GAL (10%), VIP (7%), SP (6%) or CGRP (solitary neurons), but they were SOM‐negative. No intramural neurons immunoreactive to LENK were found. The most remarkable difference in the chemical coding of enteric neurons between the control and dysenteric pigs was a very increased number of GAL‐ and VAChT‐positive nerve cells (up to 61% and 85%, respectively) in submucosal plexuses of the infected animals. The present results suggest that GAL and ACh have a specific role in local neural circuits of the inflamed porcine stomach in the course of swine dysentery.

Immunohistochemical characteristics of nerve fibres supplying the porcine vas deferens. A colocalisation study

Abstract Double-labelling immunofluorescence was used to investigate the coexistence of the catecholamine-synthesising enzymes, tyrosine hydroxylase and dopamine-β-hydroxylase and several neuropeptides including neuropeptide Y, vasoactive intestinal polypeptide, Leu5-enkephalin, somatostatin, calcitonin gene-related peptide and substance P in nerve fibres supplying the vas deferens in juvenile and adult pigs. The study has revealed three major populations of nerve terminals innervating the organ: (1) noradrenergic fibres; (2) non-noradrenergic (putative cholinergic) fibres containing vasoactive intestinal polypeptide, neuropeptide Y and somatostatin, supplying almost exclusively the lamina propria; and (3) non-noradrenergic, presumably sensory fibres, containing calcitonin gene-related peptide and substance P. The population of noradrenergic nerves can be divided into three subpopulations: a somatostatin-containing, a Leu5-enkephalin-containing and a subpopulation immunonegative to the peptides investigated, in descending order of magnitude. Coexistence patterns of the substances existing within nerve fibres supplying the vas deferens blood vessels are clearly different from those found in nerve fibres innervating the organ wall. The majority of the noradrenergic fibres associated with blood vessels contain neuropeptide Y only, while non-noradrenergic perivascular nerves contain predominantly vasoactive intestinal polypeptide. The possibility of different sources of origin of the particular nerve fibre subpopulations supplying the porcine vas deferens and its blood vessels is discussed.

Peptidergic innervation of the bovine vagina and uterus

The distribution of neuropeptide Y, substance P, vasoactive intestinal polypeptide, Leu5-enkephalin, bombesin/gastrin-releasing peptide, calcitonin gene-related peptide, somatostatin, cholecystokinin and catecholamine synthesizing enzymes, tyrosine hydroxylase and dopamine-beta-hydroxylase was studied immunohistochemically in nerve fibres supplying the bovine vagina and uterus. The nerves containing tyrosine hydroxylase or dopamine-beta-hydroxylase and neuropeptide Y-immunoreactivity were particularly numerous in both organs. Substance P, vasoactive intestinal polypeptide and Leu5-enkephalin-containing nerves were less numerous whereas somatostatin and calcitonin gene-related peptide-immunoreactive nerves occurred occasionally. Bombesin/gastrin-releasing peptide and cholecystokinin immunoreactivities were not present in nervous fibers of the bovine uterus and vagina. Generally, the immunoreactive nerve terminals, fibers, networks or nerve bundles were present below the serous membrane, between smooth muscle cells of muscular layers, around blood vessels, in the submucosal layer and below the luminal epithelium of the uterus and cervix.

Immunohistochemical Localization of Galanin in Bovine Reproductive Organs

The vagina, uterus and oviduct were shown to receive galanin immunoreactive (GAL‐IR) nerve fibres, the number of which varied between particular organs. In the ovary, GAL‐IR nerves were absent. A small number of these nerves were located in the layers of the oviduct. A moderate number of GAL‐IR nerves were situated in the body and uterine horns, whereas the uterine cervix and vagina wall contained a large number of GAL‐IR nerve fibres, evenly distributed throughout particular membranes of the organs. GAL‐IR nerves were found to contain, simultaneously, either vasoactive intestinal polypeptide (VIP), substance P (SP) or Leu5‐enkephalin (ENK). Many of the GAL‐IR nerves contained tyrosine hydroxylase (TH). A group of GAL‐IR nerves that did not possess immunoreactivity to VIP, SP, ENK or TH was also observed.