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Acrylamide is one of the harmful substances present in food. The present study aimed to establish the effect of acrylamide supplementation in tolerable daily intake (TDI) dose (0.5 µg/kg b.w./day) and a dose ten times higher than TDI (5 µg/kg b.w./day) on the population of vasoactive intestinal peptide-like immunoreactive (VIP-LI) neurons in the porcine small intestine and the degree of the co-localization of VIP with other neuroactive substances (neuronal nitric oxide synthase (nNOS), substance P (SP), and cocaine- and amphetamine-regulated transcript peptide (CART)). In our work, 15 Danish landrace gilts (5 in each experimental group) received capsules (empty or with low or high doses of acrylamide) for a period of 28 days with their morning feeding. Using double immunofluorescence staining, we established that acrylamide supplementation increased the number of neurons showing immunoreactivity towards VIP in all types of enteric nervous system (ENS) plexuses and fragments of the small intestine studied. Moreover, both doses of acrylamide led to changes in the degree of co-localization of VIP with nNOS, SP, and CART in intramural neurons. The observed changes may be the adaptation of neurons to local inflammation, oxidative stress, or the direct toxic effects of acrylamide on intestinal neurons, also referred to as neuronal plasticity.

Aspirin, also known as acetylsalicylic acid (ASA), is a commonly used anti-inflammatory drug that has analgesic and antipyretic properties. The side effects are well known, however, knowledge concerning its influence on gastric and intestinal innervation is limited. The enteric nervous system (ENS) innervates the whole gastrointestinal tract (GIT) and is comprised of more than one hundred million neurons. The capacity of neurons to adapt to microenvironmental influences, termed as an enteric neuronal plasticity, is an essential adaptive response to various pathological stimuli. Therefore, the goal of the present study was to determine the influence of prolonged ASA supplementation on the immunolocalization of neuronal nitric oxide synthase (nNOS), vasoactive intestinal peptide (VIP) and cocaine- and amphetamine- regulated transcript peptide (CART) in the porcine jejunum. The experiment was performed on 8 Pietrain × Duroc immature gilts. Using routine double-labelling immunofluorescence, we revealed that the ENS nerve cells underwent adaptive changes in response to the induced inflammation, which was manifested by upregulated or downregulated expression of the studied neurotransmitters. Our results suggest the participation of nNOS, VIP and CART in the development of inflammation and may form the basis for further neuro-gastroenterological research.

Acrylamide is one of the food toxins to which the human body is exposed. Although researchers' interest in acrylamide has been growing in recent years, the knowledge of its effect on the gastrointestinal tract, especially on intramural neurons which form the enteric nervous system is scarce. The aim of this experiment was to determine the influence of acrylamide, administered at doses equivalent to the human tolerable daily intake (TDI, 0.5 μg/kg b.w./day) and ten times higher than the TDI (5 μg/kg b.w./day), on the distribution of vasoactive intestinal peptide (VIP), substance P (SP), and calcitonin gene related peptide (CGRP) in intramural neurons of the domestic pig stomach. Using double immunofluorescent labelling we revealed that the ENS neurons underwent adaptive changes in response to the supplementation of acrylamide, which manifested themselves as increased expression of VIP, SP and CGRP, both in intramural neurons and by an increase in the nerve density in submucous and muscular layers in the porcine stomach. These substances take part in defensive reactions of neurons and transmission of sensory reactions may play an important role in protecting the stomach against the harmful effect of acrylamide. Moreover, it has been shown that acrylamide induces a significant response of ENS neurons even in TDI dose, which suggests that it is not neutral to the body. These findings may be the basis for further toxicological studies addressing the question if currently permitted minimal content of acrylamide in the food does jeopardize the health of human consumers?, (Copyright © 2018 Elsevier B.V. All rights reserved.)

Both resiniferatoxin (RTX) and tetrodotoxin (TTX) have been reported to be effective in several clinical trials aiming to cure urinary bladder dysfunction. The goal of this experiment was to study the effect of intravesical administration of RTX and TTX on the chemical coding of paracervical ganglion (PCG) neurons that supply the urinary bladder in pigs. The vasoactive intestinal peptide (VIP) and the opioid family member Leu5-enkephalin (LENK) are both known for their regulatory effects in the function of the porcine genitourinary tract. The PCG neurons innervating the urinary bladder were identified by application of the retrograde tracer Fast Blue (FB), injected into the bladder wall prior to intravesical RTX or TTX administration. Immunocytochemical detection of LENK and VIP expression in the FB-labelled perikarya revealed that in the control group 25.15% of the FB-positive PCG neurons contained LENK, and 9.22% of them expressed VIP. Intravesical infusion of RTX resulted in an increase in the number of LENKIR neurons to 48.19% and VIP-IR perikarya to 11.25%. Optional treatment with TTX induced increase of LENK-IR neurons up to 81.67% and VIP-IR population to 16.46% of the FB-positive PCG cells. The present results show that both neurotoxins affect the chemical coding of PCG nervous cells supplying the porcine urinary bladder and that they stimulate both LENK and VIP expression. Furthermore, the results indicate a possible involvement of LENK and VIP neurons in the mechanisms of action of RTX and TTX in the therapy of overactive bladder disorder.

Microplastic is an environmental hazard to which both animals and humans are exposed. Current reports show that it can cause inflammation, including in the gastrointestinal tract. To examine the impact on the ileum, 15 eight-week-old gilts (five individuals/group) were exposed to PET microplastics (7.6 µm–416.9 µm) at a dose of 0.1 g/day or 1 g/day for 28 days. The collected ileum fragments were investigated for the cytokine concentrations (IL-1β, IL-6, IL-8, IL-10, and TNF-α; ELISA test), neuron populations (cocaine and amphetamine-regulated transcript, galanin, neuronal nitric oxide synthase, substance P, vesicular acetylcholine transporter, and vasoactive intestinal peptide; immunofluorescence staining), and morphometric parameters (histological analysis). Under the influence of MP-PET, there was a reduction in the populations of CART- and GAL-positive neurons in the submucosal plexuses and of nNOS-, VAChT-, and VIP-positive neurons in all the plexuses. In contrast, there was an increase in GAL-positive neurons in the myenteric plexus and SP-positive neurons in all the plexuses. The concentrations of IL-1β, IL-6, IL-8, IL-10, and TNF-α did not undergo statistically significant changes under the influence of the low or high dose of MP-PET. The changes in the histological structure exclusively concerned the thinning of the mucosa and the muscularis externa. The results support the thesis that MP-PET is not neutral to the ileal cells. [ABSTRACT FROM AUTHOR]

Background: Currently, society and industry generate huge amounts of plastics worldwide. The ubiquity of microplastics is obvious, but its impact on the animal and human organism remains not fully understood. The digestive tract is one of the first barriers between pathogens and xenobiotics and a living organism. Its proper functioning is extremely important in order to maintain homeostasis. The aim of this study was to determine the effect of microplastic on enteric nervous system and histological structure of swine duodenum. The experiment was carried out on 15 sexually immature gilts, approximately 8 weeks old. The animals were randomly divided into 3 study groups (n = 5/group). The control group received empty gelatin capsules once a day for 28 days, the first research group received daily gelatin capsules with polyethylene terephthalate (PET) particles as a mixture of particles of various sizes (maximum particle size 300 µm) at a dose of 0.1 g/animal/day. The second study group received a dose ten times higher—1 g/animal/day. Results: A dose of 1 g/day/animal causes more changes in the enteric nervous system and in the histological structure of duodenum. Statistically significant differences in the expression of cocaine and amphetamine regulated transcript, galanin, neuronal nitric oxide synthase, substance P, vesicular acetylcholine transporter and vasoactive intestinal peptide between control and high dose group was noted. The histopathological changes were more frequently observed in the pigs receiving higher dose of PET. Conclusion: Based on this study it may be assumed, that oral intake of microplastic might have potential negative influence on digestive tract, but it is dose-dependent. [ABSTRACT FROM AUTHOR]

Environmental contamination and the resulting food contamination represent a serious problem and pose a major threat to animal and human health. The gastrointestinal tract is directly exposed to a variety of substances. One is glyphosate, whose presence in the soil is commonly observed. This study demonstrates the effects of low and high glyphosate doses on the populations of intramural neurons of the porcine descending colon. An analysis was performed on neurons ex-pressing the vasoactive intestinal peptide, pituitary adenylate cyclase-activating peptide, a neuronal isoform of nitrogen oxide synthase, and galanin. Even a low dose of glyphosate increased the number of neurons immunoreactive against the studied substances. However, the changes depended on both the plexus analysed and the substance tested. Meanwhile, a high glyphosate dose resulted in quantitative changes (an increase in the number) within neurons immunoreactive against all the studied neuropeptides/enzymes in the myenteric plexus and both submucosal plexuses. The response of the enteric nervous system in the form of an increase in the number of neurons immunoreactive against neuroprotective substances may suggest that glyphosate has a toxic effect on enteric neurons which attempt to increase their survivability through the released neuroprotective substances. [ABSTRACT FROM AUTHOR]

Simple Summary: Endometritis, both with non-infectious and infectious backgrounds, is one of the most prevalent pathological states among domestic animals. In animals, it generates severe economic problems, including lowered reproductive indices and rising medical treatment costs, and in women, it might lead to severe fertility impairment. In order to determine how the autonomic nervous system responds to such a pathological state, an experimental group of pigs were treated with Escherichia coli injection into the uterine horns, and several ganglions responsible for innervation of this organ were examined, including the paracervical ganglion located on both sides of the broad ligament of the uterus. The results clearly showed a strong impact of the inflammation on the chemical coding of neurons, some even synthesizing neurotransmitters de novo such as the GAL-expressing perikarya. Additionally, applied injections decreased the number of parasympathetic, acetylcholinesterase-expressing neurons implying the importance of the cholinergic population to keep the inflammation under control. The obtained data serve as a basis for the future implementation of modern treatment and enhancements in animal breeding. The focus of this study was based on examining the impact of endometritis on the chemical coding of the paracervical ganglion (PCG) perikaryal populations supplying pig uterus. Four weeks after the injection of Fast Blue retrograde tracer into uterine horns, either the Escherichia coli (E. coli) suspension or saline solution was applied to both horns. Laparotomy treatment was performed for the control group. Uterine cervices containing PCG were extracted on the eighth day after previous treatments. Subsequent macroscopic and histopathologic examinations acknowledged the severe form of acute endometritis in the E. coli-treated gilts, whereas double-labeling immunofluorescence procedures allowed changes to be analyzed in the PCG perikaryal populations coded with vesicular acetylcholine transporter (VAChT) and/or somatostatin (SOM), vasoactive intestinal polypeptide (VIP), a neuronal isoform of nitric oxide synthase (nNOS), galanin (GAL). The acetylcholinesterase (AChE) detection method was used to check for the presence and changes in the expression of this enzyme and further confirm the presence of cholinergic perikarya in PCG. Treatment with E. coli resulted in an increase in VAChT+/VIP+, VAChT+/VIP−, VAChT+/SOM+, VAChT+/SOM−, VAChT+/GAL− and VAChT+/nNOS− PCG uterine perikarya. An additional increase was noted in the non-cholinergic VIP-, SOM- and nNOS-immunopositive populations, as well as a decrease in the number of cholinergic nNOS-positive perikarya. Moreover, the population of cholinergic GAL-expressing perikarya that appeared in the E. coli-injected gilts and E. coli injections lowered the number of AChE-positive perikarya. The neurochemical characteristics of the cholinergic uterine perikarya of the PCG were altered and influenced by the pathological state (inflammation of the uterus). These results may indicate the additional influence of such a state on the functioning of this organ. [ABSTRACT FROM AUTHOR]

Background: The focus of the study was to examine the impact of the inflamed uterus on the population of the paracervical ganglion (PCG) uterus-innervating perikarya and their chemical coding. Fast Blue retrograde tracer was injected into the wall of uterine horns on the 17th day of the first studied estrous cycle. After 28 days, either Escherichia coli suspension or saline was applied to the horns of the uterus, whereas the control group received laparotomy only. Eight days after the above-mentioned procedures, uterine cervices with PCG were collected. Both macroscopic and histopathologic examinations confirmed severe acute endometritis in the Escherichia coli-injected uteri. The double immunofluorescence method was used to analyze changes in the PCG populations coded with dopamine-β‐hydroxylase (DβH) and/or neuropeptide Y (NPY), somatostatin (SOM), vasoactive intestinal polypeptide (VIP) and neuronal isoform of nitric oxide synthase (nNOS). Results: The use of Escherichia coli lowered the total number of Fast Blue-positive neurons. Moreover, an increase in DβH+/VIP+, DβH+/NPY+, DβH+/SOM + and DβH+/nNOS + expressing perikarya was noted. A rise in non-noradrenergic VIP-, SOM- and nNOS-immunopositive populations was also recorded, as well as a drop in DβH-positive neurotransmitter-negative neurons. Conclusions: To sum up, inflammation of the uterus has an impact on the neurochemical properties of the uterine perikarya in PCG, possibly affecting the functions of the organ. [ABSTRACT FROM AUTHOR]

Gastrointestinal inflammation resulting from prolonged NSAID drugs treatment constitutes a worldwide medical problem. The role of enteric neuroactive substances involved in this process has recently gained attention and neuropeptides produced by the enteric nervous system may play an important role in the modulation of gastrointestinal inflammation. Therefore, the aim of this study was to determine the effect of inflammation caused by indomethacin supplementation on vasoactive intestinal polypeptide (VIP), substance P (SP), neuronal nitric oxide synthase (nNOS), galanin (GAL), pituitary adenylate cyclase-activating polypeptide (PACAP), and cocaine- and amphetamine-regulated transcript peptide (CART) expression in enteric duodenal neurons in domestic pigs. Eight immature pigs of the Pietrain × Duroc race (20 kg of body weight) were used. Control animals (n=4) received empty gelatine capsules. Experimental pigs (n=4) were given indomethacin for 4 weeks, orally 10 mg/kg daily, approximately 1 h before feeding. The animals from both groups were then euthanized. Frozen sections were prepared from the collected duodenum and subjected to double immunofluorescence staining. Primary antibodies against neuronal marker PGP 9.5 and VIP, nNOS, SP, GAL, CART, and PACAP were visualized with Alexa Fluor 488 and 546. Sections were analyzed under an Olympus BX51 fluorescence microscope. Microscopic analysis showed significant increases in the number of nNOS-, VIP-, SP-, GAL-, PACAP-, and CART-immunoreactive ganglionic neurons, in both the myenteric and submucous plexuses of the porcine duodenum. The obtained results show the participation of enteric neurotransmitters in the neuronal duodenal response to indomethacin-induced inflammation. [ABSTRACT FROM AUTHOR]

Bisphenol A (BPA) is a substance used in the manufacture of plastics which shows multidirectional adverse effects on living organisms. Since the main path of intoxication with BPA is via the gastrointestinal (GI) tract, the stomach and intestine are especially vulnerable to the impact of this substance. One of the main factors participating in the regulation of intestinal functions is the enteric nervous system (ENS), which is characterized by high neurochemical diversity. Neuregulin 1 (NRG1) is one of the lesser-known active substances in the ENS. During the present study (performed using the double immunofluorescence method), the co-localization of NRG1 with other neuronal substances in the ENS of the caecum and the ascending and descending colon has been investigated under physiological conditions and after the administration of BPA. The obtained results indicate that NRG1-positive neurons also contain substance P, vasoactive intestinal polypeptide, a neuronal isoform of nitric oxide synthase and galanin and the degree of each co-localization depend on the type of enteric plexus and the particular fragment of the intestine. Moreover, it has been shown that BPA generally increases the degree of co-localization of NRG1 with other substances. [ABSTRACT FROM AUTHOR]

• VPAC1 and VPAC2 receptors are present in the porcine uteri. • Inflammation decreases VPAC1 and VPAC2 receptors mRNA expression. • After bacteria treatment, VPAC1 and VPAC2 receptors protein expression is elevated. • Inflammation decreases the contractility of uteri in response to VIP stimulation. • VPAC receptors regulate the contractility of the uterus with inflammation. Vasoactive intestinal peptide (VIP) receptor (VPAC1, VPAC2) abundances in the myometrium and functions in the regulation of inflamed uterine contractility in pigs were studied. In the CON group with gilts, only laparotomy was performed. The gilts of SAL- and E. coli -treated groups were administered saline or E. coli into the uterine horns, respectively. The E. coli -induced endometritis resulted in a lesser myometrial relative abundance of VPAC1 and VPAC2 receptor mRNA transcripts and larger abundance of protein for these receptors. In the myometrium, treatment with VIP resulted in a lesser contractility amplitude than in the tissues of the CON- and SAL- and E. coli-treated groups and in frequency in the CON- and E.coli-treated group compared to the period before VIP treatment. Compared to when there was VIP treatment alone, treatment with VPAC1 and VPAC2 receptor antagonists resulted in a lesser inhibitory effect of VIP on contractility amplitude in the myometrium of the CON and SAL-treated groups and there was complete abolishment of the inhibitory VIP effect on frequency of myometrial contractility of the CON group. In the myometrium of E. coli -treated group, treatment with VPAC1 and VPAC2 receptor antagonists resulted in a reversal of the inhibitory effect of VIP on contractility amplitude, while treatment with VPAC2 receptor antagonist resulted in elimination of contractility and a lesser endometrium/myometrium inhibitory effect of VIP on frequency of these contractions. Results indicate VIP functions to decrease myometrial contractility of the inflamed pig uterus by having functions at VPAC1 and VPAC2 receptors. [ABSTRACT FROM AUTHOR]

Simple Summary: Canine inflammatory bowel disease (IBD)—a group of gastrointestinal disorders—is a serious problem in veterinary medicine. The etiology of IBD remains unknown, and its diagnosis and effective treatment are difficult. One of the less-known aspects of IBD pathology is the influence of this disease on the enteric nervous system, which is located in the intestinal wall and regulates most of the gastrointestinal functions. Therefore, the aim of the present study was to evaluate the influence of IBD on the intramucosal nerve fibers containing vasoactive intestinal polypeptide (VIP). VIP is one of the most important substances produced by the enteric nervous structures that is involved in many regulatory processes in the gastrointestinal tract. The obtained results show that IBD induces changes in the density of intramucosal VIP-positive nerve fibers in the canine gastrointestinal tract. It suggests that VIP is involved in the pathological processes occurring during this disease. Observed changes may be a result of neuroprotective and/or adaptive processes regulated by VIP, aimed at the homeostasis maintenance in the inflamed gastrointestinal (GI) tract and induced by proinflammatory factors. Canine inflammatory bowel disease (IBD) is a group of enteropathies with nonspecific chronic symptoms and poorly understood etiology. Many aspects connected with IBD are not understood. One of them is the participation of the intestinal nervous system in the development of pathological processes. Thus, this study aimed to demonstrate changes in the density of intramucosal nerve fibers containing vasoactive intestinal polypeptide (VIP)—one of the most important intestinal nervous factors caused by the various stages of IBD development. Mucosal biopsy specimens collected from the duodenum, jejunum and descending colon of healthy dogs and dogs with varied severity of IBD were included in the experiment. The density of VIP-like immunoreactive (VIP-LI) nerves was determined by a single immunofluorescence technique and a semi-quantitative method consisting in VIP-LI fiber counts in the field of view (0.1 mm2). The obtained results indicate that IBD induces changes in the density of mucosal VIP-LI nerve fibers in the canine gastrointestinal tract. The initial decrease is followed by an increase in VIP-like immunoreactivity in successive stages of the disease. These observations show that VIP is a neuronal factor that participates in the pathological processes connected with canine IBD. The observed changes probably result from the neuroprotective and/or adaptive properties of VIP. Protective and adaptive reactions induced by inflammation aim to protect the GI tract against damage by proinflammatory factors and ensure the homeostasis in the enteric nervous system (ENS) under the conditions changed by the disease process. [ABSTRACT FROM AUTHOR]

Simple Summary: Uterine inflammation is a very frequent pathology in domestic animals leading to disturbances in reproductive processes and causing significant economic losses. The uterus possesses nerves from either the autonomic or sensory part of the peripheral nervous system. Most of the uterus-projecting neurons are localized in the caudal mesenteric ganglion. These neurons synthesize and release numerous biologically active substances in the uterus, which regulate uterine functions. The effect of inflammation on uterine innervation is poorly recognized. This study showed that Escherichia coli-induced uterine inflammation in pig led to a reduction in the total population of uterine neurons in the caudal mesenteric ganglion, and in the populations of these cells in the dorsal and central areas of this ganglion. In the caudal mesenteric ganglion of gilts after intrauterine bacterial injection, the population of uterine neurons presenting positive staining for dopamine-β-hydroxylase (an enzyme participating in noradrenaline synthesis) and negative staining for galanin, as well as the population of uterine neurons presenting negative staining for dopamine-β-hydroxylase but positive staining for neuropeptide Y, were decreased. In these gilts, there were increased numbers of uterine neurons which, besides dopamine-β-hydroxylase, also expressed neuropeptide Y, galanin and vasoactive intestinal peptide. The above changes suggest that inflammation of the gilt uterus may affect the function(s) of this organ by its action on the neurons of the caudal mesenteric ganglion. This study analyzed the influence of uterine inflammation on the neurochemical characteristics of the gilt caudal mesenteric ganglion (CaMG) uterus-supplying neurons. The horns of uteri were injected with retrograde tracer Fast Blue on day 17 of the first studied estrous cycle. Twenty-eight days later (the expected day 3 of the third studied estrous cycle), either saline or Escherichia coli suspension were administered into each uterine horn. Only the laparotomy was done in the control gilts. After 8 days, the CaMGs and uteri were harvested. The infected gilts presented a severe acute endometritis. In the CaMGs, the populations of uterine perikarya possessing dopamine-β-hydroxylase (DβH) and/or neuropeptide Y (NPY), somatostatin (SOM), galanin (GAL) and vasoactive intestinal polypeptide (VIP) were analyzed using the double immunofluorescence method. In the CaMG, bacterial injection decreased the total number of the perikarya (Fast Blue-positive), the small and large perikarya populations in the dorsal and central regions, and the small and large perikarya populations coded DβH+/GAL- and DβH-/NPY+. After bacterial treatment, there was an increase in the numbers of small and large perikarya coded DβH+/NPY+, small perikarya coded DβH+/GAL+ and DβH+/SOM- and large perikarya coded DβH+/VIP+. To summarize, uterine inflammation influences the neurochemical characteristics of the CaMG uterus-supplying neurons, which may be important for pathologically changed organ functions. [ABSTRACT FROM AUTHOR]

Hyperglycemia is one of the main causes of diabetes complications. Gastrointestinal (GI) disturbances are one of the most frequent complications during diabetes. The porcine digestive tract possesses physiological and pathological similarities to the human digestive tract. This also applies to the innervation of the gastrointestinal tract. In this study, the influence of experimentally-inducted hyperglycemia was examined on the expression of vesicular acetylcholine transporter (VAChT), cocaine- and amphetamine-regulated transcript (CART), galanin (GAL), vasoactive intestinal polypeptide (VIP), and calcitonin gene-related peptide (CGRP) in the enteric nervous system (ENS) neurons in the small intestine of the pig. During the current study, an increased number of neurons containing CART, VIP, GAL, and CGRP under streptozotocin injection were observed. The augmentation of expression included all enteric plexuses present in the small intestine. The same results were obtained in the case of VAChT; namely, chronic hyperglycemia led to an increase in the number of neurons utilizing VAChT in all investigated plexuses. The obtained results suggested that the function of neuropeptides studied in this experiment depended on their localization in the ENS structures, as well as part of the GI tract. Diabetes led to alterations in the neurochemical phenotype of small intestine enteric neurons. [ABSTRACT FROM AUTHOR]

Due to numerous therapeutic applications and high availability, non-steroidal anti-inflammatory drugs (NSAIDs) are the most widely used drugs worldwide. However, long-term use of these drugs can lead to damage to the gastrointestinal mucosa. The enteric nervous system (ENS), which is part of the autonomic nervous system, controls most aspects of gastrointestinal activity. Enteric neurons are characterized by considerable chemical plasticity and the appearance of a pathological factor results in a change in the synthesis of neurotransmitters. The purpose of this study was to determine the effects of naproxen on expression of biologically active substances by intramural neurons supplying the porcine duodenum. The study was performed on eight immature pigs of the Pietrain x Duroc race (approximately 20 kg of body weight). The animals were divided into two groups - a control (C group) and an experimental group (N group). Group C (n = 4) consisted of animals which received empty gelatine capsules. Group N (n = 4) was composed of pigs who received naproxen orally for 28 days, approximately one hour before feeding. After this time, animals from both groups were euthanized. Frozen sections (14 μm thickness) were then prepared from the collected duodenum and subjected to double immunofluorescence staining. Antibodies against the neuronal marker PGP 9.5 and against vasoactive intestinal polypeptide (VIP), substance P (SP), neuronal nitric oxide synthase (nNOS), galanin (GAL), pituitary adenylate cyclase-activating polypeptide (PACAP) and cocaine- and amphetamine- regulated transcript peptide (CART) were used as primary antibodies. The polyclonal donkey anti-rabbit, anti-mouse and anti-guinea pig IgG antibodies - Alexa Fluor 488 and 546 - were also used for staining. Analysis of the results obtained with a fluorescence microscope showed a significant increase in the number of nNOS-, VIP-, GAL-, PACAP- and CART-immunoreactive ganglionated neurons and a decrease in the number of SP-positive neurons in the myenteric and submucosal plexuses of the porcine duodenum. The obtained results indicate the participation of enteric neurotransmitters in the neuronal duodenal response to naproxen-induced inflammation. [ABSTRACT FROM AUTHOR]