Immunohistochemical study of the otic ganglion in the pig

The presence of choline acetyltransferase (ChAT), vesicular acetylcholine transporter (VAChT), neuropeptide Y (NPY), vasoactive intestinal polypeptide (VIP), somatostatin (SOM), galanin (GAL), substance P (SP) and calcitonin gene-related peptide (CGRP) was studied in neurons and nerve fibers of the porcine otic ganglion. ChAT-positive neurons were very numerous while VAChT-positive nerve cells were moderate in number. The number of neurons containing NPY and VIP was lower and those containing SOM, GAL, SP or CGRP were observed as scarce, or single nerve cells. The above mentioned substances (except SOM) were present in nerve fibers of the ganglion. ChAT- and VAChT-positive nerve fibers were numerous, while the number of nerve terminals containing NPY, VIP and SP was lower. GAL- and CGRP-positive nerve fibers were scarce.     

Immunohistochemical characteristics of neurons supplying the porcine bulbourethral gland

In the male pig, the bulbourethral gland (BG) is a particulary well developed accessory genital gland (AGG) which produces complex secretion contributing to the fluid component of semen. The secretory and motor function of AGGs is thought to be under the autonomic nervous system control. Although relatively much is known about the innervation of the prostate gland and, to a lesser degree, of the seminal vesicle, the paucity of data dealing with the innervation of BG is striking. Therefore, combined retrograde tracing and double-labelling immunofluorescence have been used to investigate the distribution and immunohistochemical properties of autonomic and primary afferent neurons projecting to this gland in the pig. BG-projecting neurons were found in some ipsilateral (I) and contralateral (C) sympathetic chain ganglia (SChG), the caudal mesenteric ganglion (CaMG), pelvic ganglia (PG) and some dorsal root ganglia (DRG). Immunohistochemistry revealed that the vast majority of CaMG and SChG BG-projecting neurons contained tyrosine hydroxylase (TH) and dopaminebeta-hydroxylase (DbetaH), and some neuropeptides including neuropeptide Y (NPY), somatostatin (SOM) and galanin (GAL). Three subpopulations of PG neurons supplying BG could be distinguished: 1) cholinergic neurons [vesicular acetylcholine transporter (VAChT)-positive] which also contained vasoactive intestinal polypeptide (VIP), nitric oxide synthase (NOS), SOM and NPY, 2) adrenergic neurons (TH-positive) which also stained for NPY, GAL or leu5-enkephalin (LEU), and 3) non-adrenergic, non-cholinergic neurons (NANC). DRG BG-projecting neurons contained mostly substance P (SP) and/or calcitonin gene-related peptide (CGRP) which sometimes colocalized with GAL. The possible functional significance of the substances found within the neurons is discussed.     

Immunohistochemical characterization of neurons in the porcine ciliary ganglion

The present study was aimed at disclosing the chemical coding of nerve structures in the porcine ciliary ganglion (CG) using immunohistochemical methods. The substances under investigation included markers of "classical" neurotransmitters, choline acetyltransferase (ChAT), vesicular acetylcholine transporter (VAChT), tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DbetaH) as well as neuropeptides, somatostatin (SOM), galanin (GAL), substance P (SP), calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide (VIP) and neuropeptide Y (NPY). Immunoreactivity to ChAT and VAChT was found virtually in all the neuronal somata and in numerous intraganglionic, varicose nerve fibres which often formed basket-like formations around the nerve cell bodies. Many CG neurons contained immunoreactivity for SOM (46%) or GAL (29%). Interestingly, a small number (approx. 1%) of the cholinergic somata stained for TH but not for DbetaH; nevertheless, some extra- and intraganglionic nerve fibres displayed immunoreactivity for DbetaH or TH. The CG perikarya stained neither for vasoactive intestinal polypeptide (VIP) nor for neuropeptide Y (NPY), but some NPY- or VIP-positive nerve terminals were observed within nerve bundles distributed outside the ganglion. SP- and CGRP-immunoreactivity was found in some intraganglionic nerve fibres only. The present study revealed that the porcine CG consists of cholinergic neurons many of which contain SOM and GAL. Thus, it can be assumed that in the pig, these neuropeptides are involved, complementary to acetylocholine, in the parasympathetic postganglionic nerve pathway to structures of the eye including the ciliary and iris sphincter muscles.     

The Distribution and Chemical Coding of Intramural Nerve Structures Supplying the Porcine Stomach

The present study investigated the arrangement and chemical coding of intramural nerve structures supplying the porcine stomach. Tissue samples comprising all layers of the wall of the ventricular fundus were collected from juvenile female pigs (n = 4), which were first deeply anaesthetized and then transcardially perfused with buffered paraformaldehyde. 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 neurochemical characteristics using antibodies against vesicular acetylcholine transporter (VAChT), nitric oxide synthase (NOS), galanin (GAL), vasoactive intestinal‐polypeptide (VIP), somatostatin (SOM) and substance P (SP). The study confirmed the presence of three distinct nerve plexuses within the wall of the porcine stomach including one myenteric plexus and two, outer and inner, submucous plexuses. The outer and inner submucous plexuses (OSP and ISP, respectively) were similar in respect to the chemical coding of neurons they contained. Most of the neurons expressed immunoreactivity to SP (ISP 58%; OSP 60%) or to VAChT (ISP 56%; OSP 56%), some of them stained for GAL (ISP 18%; OSP 15%) and solitary nerve cells were SOM‐positive (in ISP only). No neurons in the submucous plexuses displayed immunoreactivity to VIP or NOS. In the myenteric plexus, some neurons stained for NOS (20%), VAChT (15%), GAL (10%), VIP (8%) or SP (8%) while no neurons immunoreactive for SOM were encountered. In both submucous and myenteric plexuses, many varicose nerve fibres expressed immunoreactivity to VAChT, GAL or SP, while VIP‐, SOM‐ or NOS‐positive nerve terminals were less numerous. The comparison of the present results with those obtained by other authors has revealed distinct inter‐species differences regarding the arrangement and chemical coding of nerve structures supplying the mammalian stomach.     

Effect of total or partial uterus extirpation on sympathetic uterus-projecting neurons in porcine inferior mesenteric ganglion. B. Changes in expression of neuropeptide Y,galanin, vasoactive intestinal polypeptide,pituitary adenylate-cyclase activating peptide,somatostatin and substance P

The expression of neuropeptide Y (NPY), galanin (GAL), vasoactive intestinal polypeptide (VIP), pituitary adenylate cyclase-activating peptide (PACAP), somatostatin (SOM) and substance P (SP) was studied in the neurons of the inferior mesenteric ganglion (IMG) projecting to the uterine horn and uterine cervix after uterus extirpation-induced axotomy in sexually immature gilts. The expression was studied with immunohistochemistry, in situ hybridization and RT-PCR. Uterus-projecting neurons were identified by retrograde tracing with Fast Blue (FB). Immunohistochemistry revealed that FB-positive (FB+) uterus-projecting neurons in control animals contained only immunoreactivities to NPY (ca. 50%) and GAL (single neurons). Uterus extirpation increased the occurrence of NPY and GAL in FB+ neurons. No other studied neuropeptides were found in axotomized uterus-projecting neurons. Hybridization in situ revealed the reduction of NPY expression and induction of GAL expression in FB+ neurons. RT-PCR detected induction of GAL expression in the IMG after uterus extirpation. The expression level of NPY and SOM was significant and was not affected by axotomy. The expression level of PACAP was very low and did not differ between IMG of control, partially and totally hysterectomized animals. No VIP and SP expression was detected in all ganglia. The presented data show clear axotomy-related changes in the expression of GAL and NPY in the uterus-projecting neurons of the porcine IMG.     

Immunohistochemical Characterization of Neurones in the Hypoglossal Nucleus of the Pig

With 4 figures and 1 table In this study, the presence of several neurotransmitters and transmitter synthesizing enzymes was studied in hypoglossal nucleus (HN) of the juvenile (4 months old) female pigs (n = 3). Double‐labeling immunofluorescence revealed neurones expressing cholinacetyltranspherase (ChAT), calcitonin gene‐related peptide (CGRP), nitric oxide synthase (NOS), and somatostatin (SOM). Nerve fibers within HN were ChAT, CGRP, NOS, SOM, substance P (SP), Leu‐5‐enkephalin (Leu‐5‐Enk), ß‐dopamine hydroxylase (DßH), neuropeptide Y (NPY) positive. Virtually all the perikarya contained ChAT, whereas CGRP was present in 47% of the neurones. Nerve cell bodies containing NOS or SOM were only occasionally observed. Immunoreactive nerve fibers were found in a close vicinity of the perikarya, often forming baskets around nerve cell bodies. The results obtained were compared with similar data obtained in other species. The presence of immunoreactive structures, origin of the nerve fibers, and functional significance of the findings are discussed.     

Expression of neuronal nitric oxide synthase in the pancreas of the sheep

In numerous mammals, nitric oxide (NO) influences the activity of the exocrine and endocrine pancreas. In this study, immunocytochemistry was utilized to investigate the expression of neuronal nitric oxide synthase (nNOS) in the pancreas of sheep. In double immunocytochemical staining, the co-localization of nNOS with vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY) or substance P (SP) was studied. The presence of nNOS was confined to the intrapancreatic neurones (9.6 +/- 1.3%) as well as to nerve fibres of the endocrine pancreas and intrapancreatic ganglia. nNOS-immunoreactive (IR) neurones were round and oval in shape and predominantly (83.3 +/- 2.6%) belonged to the middle-size group (25-50 mum). Numerous, fine islets supplying nNOS-IR nerve terminals were devoid of VIP, SP or NPY. Moderately numerous, non-varicose nNOS-IR nerve fibres of intrapancreatic ganglia frequently expressed VIP or NPY, but not SP; 2.2 +/- 0.6% of nNOS-IR intrapancreatic neurones displayed lack of VIP, whereas 7.5 +/- 0.8% were VIP-IR. All nNOS-IR neurones were devoid of SP. The frequencies of nNOS-IR/NPY-IR and nNOS-IR/NPY-negative intrapancreatic neurones were 2.2 +/- 0.4% and 6.1 +/- 1.1%, respectively. Comparison with other mammals indicated that nitrergic innervation of the ovine pancreas is species-determined and may be a reflection of the ruminants' digestion specificity. The possible origin of nNOS-IR nerve fibres and functional significance of NO in the pancreas of sheep were discussed.     

Vasculogenesis by Endothelial Progenitor Cells In Vitro

The present study investigated the arrangement and chemical coding of intramural nerve structures supplying the porcine stomach. Tissue samples comprising all layers of the wall of the ventricular fundus were collected from juvenile female pigs ( n  = 4), which were first deeply anaesthetized and then transcardially perfused with buffered paraformaldehyde. 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 neurochemical characteristics using antibodies against vesicular acetylcholine transporter (VAChT), nitric oxide synthase (NOS), galanin (GAL), vasoactive intestinal-polypeptide (VIP), somatostatin (SOM) and substance P (SP). The study confirmed the presence of three distinct nerve plexuses within the wall of the porcine stomach including one myenteric plexus and two, outer and inner, submucous plexuses. The outer and inner submucous plexuses (OSP and ISP, respectively) were similar in respect to the chemical coding of neurons they contained. Most of the neurons expressed immunoreactivity to SP (ISP 58%; OSP 60%) or to VAChT (ISP 56%; OSP 56%), some of them stained for GAL (ISP 18%; OSP 15%) and solitary nerve cells were SOM-positive (in ISP only). No neurons in the submucous plexuses displayed immunoreactivity to VIP or NOS. In the myenteric plexus, some neurons stained for NOS (20%), VAChT (15%), GAL (10%), VIP (8%) or SP (8%) while no neurons immunoreactive for SOM were encountered. In both submucous and myenteric plexuses, many varicose nerve fibres expressed immunoreactivity to VAChT, GAL or SP, while VIP-, SOM- or NOS-positive nerve terminals were less numerous. The comparison of the present results with those obtained by other authors has revealed distinct inter-species differences regarding the arrangement and chemical coding of nerve structures supplying the mammalian stomach.     

Analysis of the chemical coding of neurons in the intermediate thoracic ganglion of the pig

The pig has been widely used as a model in cardiovascular research. A unique feature of the porcine extrinsic sympathetic cardiac nerves is that they arise from intermediate ganglia in the thoracic cavity. The localization and pattern of distribution of nerve cell bodies and fibers containing tyrosine hydroxylase (TH), dopamine B-hydroxylase (DBH), neuropeptide Y (NPY), vasoactive intestinal polypeptide (VIP), somatostatin (SOM), galanin (GAL), methionine-enkephalin (MET) as well as calcitonin gene-related peptide (CGRP), substance P (SP) and pituitary adenylate cyclase-activating peptide (PACAP) was studied with immunohistochemistry. Almost all the neurons showed immunoreactivity to TH. Immunoreactivity to NPY, VIP, SOM, GAL, MET and PACAP was displayed by nerve cell bodies while nerve fibers exhibited immunoreactivity to all the neuropeptides studied. Therefore, it seems that the chemical coding of neurons and especially nerve fibers in the porcine intermediate ganglion share general similarities (with certain neurochemical variability), with porcine prevertebral ganglia (e.g., celiacomesenteric and caudal mesenteric ganglia).     

The influence of experimental ileitis on the neuropeptide coding of enteric neurons in the pig

In the present study, both the ELISA test and immunohistochemical staining were used to investigate the influence of artificially induced ileitis on the chemical coding of enteric neurons in the pig. The ileum wall in experimental (E) pigs was injected in multiple sites with 4% paraformaldehyde to induce inflammation, while in the control (C) animals, the organ was injected with 0.1M phosphate buffer (pH 7.4). Three days after ileitis induction, samples of ileum wall from all the animals were evaluated for VIP, SP, CGRP, NPY, GAL and SOM concentration (ELISA test) and the expression of these biologically active substances by the enteric neurons (immunohistochemical staining). Quantitative results showed that ileitis decreased tissue concentration of VIP, CGRP and SOM but increased tissue concentration of SP, NPY and GAL. Immunochemistry revealed that in both the experimental and control pigs, VIP-positive (VIP+) nerve fibers supplied mainly ileal blood vessels, and the labeled pericarya were located in the inner (ISP) and outer submucous plexus (OSP). SP+ and CGRP+ nerve terminals were found in both the mucous and muscular membrane, while the labeled pericarya were found in ISP, OSP and myenteric plexus (MP). In both C and E pigs, the very few nerve terminals containing NPY and SOM were located mainly in the mucous membrane. NPY- or/and SOM-immunopositive nerve cell bodies were found in ISP, OSP and MP. GAL+ nerve fibers supplied all layers of the ileum and were most numerous in the muscular membrane, while the labeled pericarya were present in all the enteric plexuses. The present results suggest that enteric neurons are highly plastic in their response to inflammation.     

New aspects in the pathogenesis of abomasal displacement

Impaired abomasal motility and an increased accumulation of gas are prerequisites for displacement of the abomasum in the cow. Predisposing factors are the breed (e.g. Holstein-Friesian, Simmental-Red-Holstein cross breeds and Guernsey), genetic background, twin pregnancy, first weeks of lactation, metabolic disorders (ketosis, increased lipomobilisation, insulin resistance), high-concentrate and low-fibre diets, as well as other concomitant diseases, such as endometritis, mastitis and claw disorders. There does not appear to be a strong correlation between increased milk yield or endotoxaemia and abomasal displacement. Recent studies have focused on possible functional disorders of the enteric nervous system within the abomasal wall, since cattle with abomasal displacement have an increased activity of neuronal nitric oxide synthase, as well as decreased acetylcholine sensitivity. In addition, there appear to be significant differences between breeds in the levels of the neurotransmitters substance P (SP) and vasoactive intestinal peptide (VIP) in the abomasal wall. For example, SP (stimulatory) was significantly less in German Holsteins in comparison to the German Fleckvieh, whereas VIP (inhibitory) was markedly increased. These risk factors may explain why Holstein cows are more susceptible to abomasal displacement than other breeds.     

Peptidergic and nitrergic innervation of the pineal gland in the domestic pig: an immunohistochemical study

The presence and co-localization of vasoactive intestinal polypeptide (VIP), peptide N-terminal histidine C-terminal isoleucine (PHI), pituitary adenylate cyclase-activating peptide (PACAP), somatostatin (SOM), calcitonin gene-related peptide (CGRP), substance P (SP) and the neuronal isoform of nitric oxide synthase (NOS) were studied in neuronal structures of the pig pineal gland. Paraformaldehyde-fixed pineals of 3-month-old gilts were sliced into serial cryostat sections, which were subjected to a set of double immunofluorescence stainings. Based on the co-existence patterns of neuropeptides, five populations of nerve fibres supplying the pig pineal were distinguished: (1) PHI-positive, (2) PACAP-positive, (3) SOM-positive, (4) SP/CGRP-positive and (5) SP-positive/CGRP-negative. Only a subpopulation of PHI-positive fibres contained VIP at the level detectable by immunofluorescence. NOS was found in some intrapineal PHI- and VIP-positive fibres. PHI-, VIP- and NOS-positive nerve fibres were more numerous in the peripheral than in the central part of the pineal. PACAP-positive fibres were equally distributed within the gland. The density of SOM-positive fibres was higher in the ventro-proximal than in the dorso-distal part of the pineal. SOM was also detected in some neuronal-like cells or specialized pinealocytes situated in the central region of the gland. Two populations of fibres containing SP were found: CGRP-positive, present in the distal and central parts of the pineal as well as CGRP-negative, localized in the proximal compartment of the gland.     

An immunohistochemical study of various peptide-containing endocrine cells and neurones at the equine ileocaecal junction

The ileocaecal junctions of 5 horses and 2 donkeys were examined by using antisera to the following peptides: somatostatin, glucagon, gastrin, neurotensin, vasoactive intestinal peptide (VIP), peptide histidine isoleucine (PHI), calcitonin gene-related peptide (CGRP), substance P (SP) and neuropeptide Y (NPY). Antisera to somatostatin, neurotensin and NPY demonstrated endocrine cells in the ileal- and caecal parts of the ileocaecal junction, while immunoreactivity for glucagon was demonstrated in endocrine cells of the ileal part only. Nerve cell bodies showing immunoreactivity to SP, VIP, CGRP and PHI were demonstrated in the myenteric and submucosal plexuses and were associated with small blood vessels in the submucosa of all the regions tested. Ramified nerve fibres in the submucosa immunoreactive to SP, VIP, CGRP and PHI extended to the mucosa and to small blood vessels in the submucosa. Nerve fibres showing immunoreactivity to SP, VIP and PHI extended to the circular smooth muscle layer of the ileocaecal junction.     

Immunohistochemical study of the pre- and postnatal innervation of the dog lower urinary tract: morphological aspects at the basis of the consolidation of the micturition reflex

Immunohistochemical studies were performed on male and female bladder and urethra collected from 4 adults dogs and 10 foetal specimens with crown-rump length from 53 to 155 mm (medium-sized breeds, presumptive 38 days of gestation to term). A panel of antisera was tested, including PGP 9.5 to describe the general intramural innervation, ChAT and TH to depict the cholinergic and nor-adrenergic components and NOS1, CGRP, SP, NPY, VIP, SOM, GAL, 5-HT to investigate the possible nitrergic, peptidergic and aminergic ones. A rich cholinergic innervation was present in adult bladder and urethra, along with a lesser number of adrenergic nerves and a small number of nitrergic ones. Either bladder or urethra received numerous CGRP-, SP-, NPY-, VIP-containing nerve fibres which were distributed throughout the muscle layers. All over the lower urinary tract strong to weak ChAT-, CGRP-, SP- and NPY-immunoreactivity was detected in intramural ganglia, in peripheral nerve bundles and around blood vessels. 5-HT-immunoreactive endocrine cells were present in the urethral epithelium. Early foetal organs were supplied only by cholinergic nerve fibres. Few NOS-, CGRP- and SP-ergic components appeared at the end of pregnancy. It can be guessed that sensory mediators such as CGRP and SP increase in postnatal ages while other neuropeptides, such as NPY and VIP, appear only after birth, as the urinary reflex consolidates.     

Chemical Coding of Sensory Neurons Supplying the Hip Joint Capsule in the Sheep

Immunohistochemical properties of nerve fibres supplying the joint capsule were previously described in many mammalian species, but the localization of sensory neurons supplying this structure was studied only in laboratory animals, the rat and rabbit. However, there is no comprehensive data on the chemical coding of sensory neurons projecting to the hip joint capsule (HJC). The aim of this study was to establish immunohistochemical properties of sensory neurons supplying HJC in the sheep. The study was carried out on 10 sheep, weighing about 30–40 kg. The animals were injected with a retrograde neural tracer Fast Blue (FB) into HJC. Sections of the spinal ganglia (SpG) with FB‐positive (FB+) neurons were stained using antibodies against calcitonin gene‐related peptide (CGRP) substance P (SP), pituitary adenylate cyclase‐activating peptide (PACAP), nitric oxide synthase (n‐NOS), neuropeptide Y (NPY), vasoactive intestinal polypeptide (VIP), Leu‐5‐enkephalin (Leu‐Enk), galanin (GAL) and vesicular acetylcholine transporter (VACHT). The vast majority of FB+ neurons supplying HJC was found in the ganglia from the 5th lumbar to the 2nd sacral. Immunohistochemistry revealed that most of these neurons were immunoreactive to CGRP or SP (80.7 ± 8.0% or 56.4 ± 4.8%, respectively) and many of them stained for PACAP or GAL (52.9 ± 2.9% or 50.6 ± 19.7%, respectively). Other populations of FB+ neurons were those immunoreactive to n‐NOS (37.8 ± 9.7%), NPY (34.6 ± 6.7%), VIP (28.7 ± 4.8%), Leu‐Enk (27.1 ± 14.6) and VACHT (16.7 ± 9.6).     

Immunohistochemical Study of Peptide-containing Nerves in the Gastrointestinal Tract of the Japanese Field Vole, Microtus montebelli

The distribution and relative frequency of nerves containing immunoreactivity for substance P (SP), vasoactive intestinal polypeptide (VIP), gastrin-releasing polypeptide (GRP), calcitonin gene-related peptide (CGRP), neuropeptide Y (NPY) and menthionine-enkephalin (MENK) were studied by immunohistochemistry in the gastrointestinal tract of the herbivorous Japanese field vole, Microtus montebelli.     

Role of endothelium and nitric oxide in modulating in vitro responses of colonic arterial and venous rings to vasodilatory neuropeptides in horses

The objective of this study was to determine and compare the in vitro responses of equine large colon arterial and venous rings to vasodilatory neuropeptides; calcitonin gene-related peptide (CGRP); substance P (SP); vasoactive intestinal polypeptide (VIP); and acetylcholine (ACh), a standard nonpeptide endothelium-dependent vasodilator. Responses of vessel rings to graded concentrations (10(-11) M to 10(-5) M) of each drug were determined in endothelium-intact, denuded, and Nomega-nitro-L-arginine methyl ester (L-NAME, 10(-5) M)-treated rings that were pre-contracted with norepinephrine. Percentage maximal relaxation (PMR), defined as the % decrease from the contracted state, was determined. Because all rings did not relax at least 50%, EC50 values could not be consistently calculated. Arterial rings with intact endothelium were more sensitive to CGRP, compared with VIP and SP, and venous rings of all conditions were more sensitive to VIP than CGRP or SP. Overall, arteries had a greater PMR for ACh compared with SP and VIP. Intact and L-NAME treated arteries had a greater PMR than denuded arteries; there were no differences in PMR of intact and L-NAME treated arteries. Veins had a greater PMR for VIP than CGRP, SP, or ACh. Calcitonin gene-related peptide caused greater relaxation in intact arteries, whereas VIP causes greater relaxation in veins. Arterial relaxation was dependent upon the presence of intact endothelium. The response of veins to VIP among the conditions tested was not different, suggesting VIP has direct actions on venous smooth muscle. These neuropeptides modulate vasomotor tone via vasorelaxation in colonic arteries and veins.     

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), Leu(5)-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.