Glyphosate-induced changes in the expression of galanin and GALR1, GALR2 and GALR3 receptors in the porcine small intestine wall.

Glyphosate is the active ingredient of glyphosate-based herbicides and the most commonly used pesticide in the world. The goal of the present study was to verify whether low doses of glyphosate (equivalent to the environmental exposure) evoke changes in galanin expression in intramural neurons in the small intestine in pigs and to quantitatively determine changes in the level of galanin receptor encoding mRNA (GALR1, GALR2, GALR3) in the small intestine wall. The experiment was conducted on 15 sexually immature gilts divided into three study groups: control (C)-animals receiving empty gelatin capsules; experimental 1 (G1)-animals receiving a low dose of glyphosate (0.05 mg/kg b.w./day); experimental 2 (G2)-animals receiving a higher dose of glyphosate (0.5 mg/kg b.w./day) orally in gelatine capsules for 28 days. Glyphosate ingestion led to an increase in the number of GAL-like immunoreactive intramural neurons in the porcine small intestine. The results of RT-PCR showed a significant increase in the expression of mRNA, which encodes the GAL-receptors in the ileum, a decreased expression in the duodenum and no significant changes in the jejunum. Additionally, intoxication with glyphosate increased the expression of SOD2-encoding mRNA in the duodenum and decreased it in the jejunum and ileum, but it did not affect SOD1 expression. The results suggest that it may be a consequence of the cytotoxic and/or neurotoxic properties of glyphosate and/or its ability to induce oxidative stress.

Changes in the Phenotype of Intramural Inhibitory Neurons of the Porcine Descending Colon Resulting from Glyphosate Administration.

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.

Effect of different glyphosate doses on the chemical coding of neurons of the enteric nervous system of the porcine descending colon.

BACKGROUND: Neurons of the enteric nervous system are characterised by high neuronal plasticity, with their number likely to change in response to various endogenous and exogenous substances. MATERIALS AND METHODS: Fifteen sexually immature gilts divided into 3 groups were used: control - animals receiving empty gelatin capsules; G1 - animals receiving a low dose of glyphosate - 0.05 mg/kg bw/day; G2 - animals receiving a higher dose of glyphosate-0.5 mg/kg/day in gelatin capsules orally for 28 days. Frozen sections were then subjected to the procedure of double immunofluorescent staining. RESULTS: With low-dose supplementation, no effect on the SP- and CART-positive neuron population was observed. However, a reduction in the number of VAChT-positive neurons in the internal submucosal plexus was described, while the number of CGRP-positive neurons increased in all enteric plexuses. In response to a high glyphosate dose, the quantitative variability of the neurons was significantly more pronounced than that for a low dose. There was an increase in the number of SP- and CGRP-positive neurons and a decrease in the number of VAChT-positive neurons in both the myenteric plexus and the submucosal plexuses. The response of CART-positive neurons was the weakest, as a high dose of glyphosate led to an increase in the number of neurons only in the myenteric plexus. CONCLUSIONS: The above data show that glyphosate is an exogenous substance that affects neuronal populations of the enteric nervous system, in this case, the descending colon.

Acrylamide-Induced Changes in the Pituitary Adenylate Cyclase-Activating Polypeptide (PACAP) Immunoreactivity in Small Intestinal Intramural Neurons in Pigs.

BACKGROUND: A particularly pressing problem is determining consumer-safe doses of potentially health- and life-threatening substances, such as acrylamide. The aim of the study was to determine how acrylamide affects the pituitary adenylate cyclase-activating polypeptide (PACAP)-immunoreactive intramural neurons in the small intestine of sexually immature gilts. METHODS: The study was conducted on 15 sexually immature Danish gilts receiving for 28 days empty gelatin capsules or acrylamide in low (0.5 µg/kg of body weight (b.w.)/day) and high (5 µg/kg b.w./day) doses. After euthanasia, intestinal sections were stained using the double immunofluorescence staining procedure. RESULTS: Studies have shown that oral administration of acrylamide in both doses induced a response of intramural neurons expressed as an increase in the population of PACAP-immunoreactive neurons in the small intestine. In the duodenum, only in the myenteric plexus (MP) was an increase in the number of PACAP-immunoreactive (IR) neurons observed in both experimental groups, while in the outer submucous plexus (OSP) and inner submucous plexus (ISP), an increase was noted only in the high-dose group. In the jejunum, both doses of acrylamide led to an increase in the population of PACAP-IR neurons in each enteric plexus (MP, OSP, ISP), while in the ileum, only supplementation with the higher dose of acrylamide increased the number of PACAP-IR enteric neurons in the MP, OSP, and ISP. CONCLUSIONS: The obtained results suggest the participation of PACAP in acrylamide-induced plasticity of enteric neurons, which may be an important line of defence from the harmful action of acrylamide on the small intestines.

Glyphosate affects the neurochemical phenotype of the intramural neurons in the duodenum in the pig.

BACKGROUND: Glyphosate-based herbicides have been one of the most intensively used pollutants worldwide and food products containing glyphosate are an essential component of human and animal diet. The aim of present study was to determine the effect of glyphosate intoxication on the neurochemical properties of the enteric nervous system (ENS) neurons located in the wall of the porcine duodenum. METHODS: Fifteen sexually immature gilts divided into 3 groups were used: control-animals receiving empty gelatin capsules; G1-animals receiving a low dose of glyphosate-corresponding to the theoretical maximum daily intake (TMDI) - 0.05 mg/kg bw/day; G2-animals receiving a higher dose of glyphosate-corresponding to the acceptable daily intake (ADI)-0.5 mg/kg/day in gelatin capsules orally for 28 days. After this time, the animals were euthanized and small intestine samples were collected. Frozen sections were then subjected to the procedure of double immunofluorescent staining. KEY RESULTS: Glyphosate supplementation led to alterations in the neurochemical code of the ENS neurons in the porcine duodenum. Generally, increased population of neurons immunoreactive to PACAP, CGRP, CART, nNOS, and a decreased number of VAChT-like immunoreactive neurons were noted. CONCLUSIONS AND INFERENCES: It may be a first preclinical symptom of digestive tract dysfunction in the course of glyphosate intoxication and further studies are needed to assess the toxicity and risks of glyphosate to humans.

Involvement of the calcitonin gene-related peptide system in the modulation of inflamed uterus contractile function in pigs.

This study analyzed severe acute endometritis action on myometrial density and distribution of protein gene product (PGP)9.5- and calcitonin gene-related peptide (CGRP)-like immunoreactive nerve fibers and calcitonin receptor-like receptor (CLR) expression, and on CGRP receptor (CGRPR) participation in uterine contractility in pigs. E. coli suspension (E. coli group) or saline (SAL group) were injected into the uteri, or only laparotomy was performed (CON group). In the E. coli group myometrium, a lack of significant changes in PGP9.5 and CGRP innervation patterns and increased CLR protein level were revealed. In all groups, compared to the pretreatment period, human αCGRP increased amplitude in the myometrium, while reducing it in endometrium/myometrium. In the E. coli group endometrium/myometrium, human αCGRP lowered amplitude vs other groups. Human αCGRP reduced frequency in CON and SAL groups and enhanced it in the E. coli group endometrium/myometrium. The frequency in E. coli group increased vs other groups. CGRPR antagonist, human αCGRP8-37, reversed (CON, SAL groups) and eliminated (E. coli group) the rise in human αCGRP-induced myometrial amplitude. In endometrium/myometrium, human αCGRP8-37 abolished (CON group) and reversed (SAL group) a decrease in frequency, and reduced the rise in frequency (E. coli group) caused by human αCGRP. Collectively, in the myometrium, endometritis did not change PGP9.5 and CGRP innervation patterns and enhanced CLR protein level. CGRPR also mediated in CGRP action on inflamed uterus contractility.

Administration of Different Doses of Acrylamide Changed the Chemical Coding of Enteric Neurons in the Jejunum in Gilts.

Excessive consumption of highly processed foods, such as chips, crisps, biscuits and coffee, exposes the human to different doses of acrylamide. This chemical compound has a multidirectional, adverse effect on human and animal health, including the central and peripheral nervous systems. In this study, we examined the effect of different doses of acrylamide on the enteric nervous system (ENS) of the porcine jejunum. Namely, we took into account the quantitative changes of neurons located in the jejunum wall expressing substance P (SP), galanin (GAL), a neuronal form of nitric oxide synthase (nNOS), the vesicular acetylcholine transporter (VAChT) and cocaine- and amphetamine-regulated transcript (CART). The obtained results indicate that acrylamide causes a statistically significant increase in the number of neurons immunoreactive to SP, GAL, VAChT and CART in all types of examined enteric plexuses and a significant drop in the population of nNOS-positive enteric neurons. Changes were significantly greater in the case of a high dose of acrylamide intoxication. Our results indicate that acrylamide is not indifferent to ENS neurons. A 28-day intoxication with this substance caused marked changes in the chemical coding of ENS neurons in the porcine jejunum.

Streptozotocin-Induced Diabetes Causes Changes in Serotonin-Positive Neurons in the Small Intestine in Pig Model.

Serotonin (5-hydroxytryptamine or 5-HT) is an important neurotransmitter of the central and peripheral nervous systems, predominantly secreted in the gastrointestinal tract, especially in the gut. 5-HT is a crucial enteric signaling molecule and is well known for playing a key role in sensory-motor and secretory functions in the gut. Gastroenteropathy is one of the most clinical problems in diabetic patients with frequent episodes of hyperglycemia. Changes in 5-HT expression may mediate gastrointestinal tract disturbances seen in diabetes, such as nausea and diarrhea. Based on the double immunohistochemical staining, this study determined the variability in the population of 5-HT-positive neurons in the porcine small intestinal enteric neurons in the course of streptozotocin-induced diabetes. The results show changes in the number of 5-HT-positive neurons in the examined intestinal sections. The greatest changes were observed in the jejunum, particularly within the myenteric plexus. In the ileum, both de novo 5-HT synthesis in the inner submucosal plexus neurons and an increase in the number of neurons in the outer submucosal plexus were noted. The changes observed in the duodenum were also increasing in nature. The results of the current study confirm the previous observations concerning the involvement of 5-HT in inflammatory processes, and an increase in the number of 5-HT -positive neurons may also be a result of increased concentration of the 5-HT in the gastrointestinal tract wall and affects the motor and secretory processes, which are particularly intense in the small intestines.

Pituitary Adenylate Cyclase-Activating Polypeptide (PACAP) in Physiological and Pathological Processes within the Gastrointestinal Tract: A Review.

Pituitary adenylate cyclase-activating polypeptide (PACAP) is a neuropeptide widely distributed in the central nervous system (CNS) and many peripheral organs, such as the digestive tract, endocrine, reproductive and respiratory systems, where it plays different regulatory functions and exerts a cytoprotective effect. The multifarious physiological effects of PACAP are mediated through binding to different G protein-coupled receptors, including PAC1 (PAC1-R), VPAC1 (VPAC1-R) and VPAC2 (VPAC2-R) receptors. In the gastrointestinal (GI) tract, PACAP plays an important regulatory function. PACAP stimulates the secretion of digestive juices and hormone release, regulates smooth muscle contraction, local blood flow, cell migration and proliferation. Additionally, there are many reports confirming the involvement of PACAP in pathological processes within the GI tract, including inflammatory states, neuronal injury, diabetes, intoxication and neoplastic processes. The purpose of this review is to summarize the distribution and pleiotropic action of PACAP in the control of GI tract function and its cytoprotective effect in the course of GI tract disorders.

Diabetes Affects the Pituitary Adenylate Cyclase-Activating Polypeptide (PACAP)-Like Immunoreactive Enteric Neurons in the Porcine Digestive Tract.

Diabetic gastroenteropathy is a common complication, which develops in patients with long-term diabetes. The pituitary adenylate cyclase-activating polypeptide (PACAP) is a neuropeptide known for its cytoprotective properties and plays an important role in neuronal development, neuromodulation and neuroprotection. The present study was designed to elucidate, for the first time, the impact of prolonged hyperglycaemia conditions on a population of PACAP-like immunoreactive neurons in selected parts of the porcine gastrointestinal tract. The experiment was conducted on 10 juvenile female pigs assigned to two experimental groups: The DM group (pigs with streptozocin-induced diabetes) and the C group (control pigs). Diabetes conditions were induced by a single intravenous injection of streptozocin. Six weeks after the induction of diabetes, all animals were euthanised and further collected, and fixed fragments of the stomach, duodenum, jejunum, ileum and descending colon were processed using the routine double-labelling immunofluorescence technique. Streptozotocin-induced hyperglycaemia caused a significant increase in the population of PACAP-containing enteric neurons in the porcine stomach, small intestines and descending colon. The recorded changes may result from the direct toxic effect of hyperglycaemia on the ENS neurons, oxidative stress or inflammatory conditions accompanying hyperglycaemia and suggest that PACAP is involved in regulatory processes of the GIT function in the course of diabetes.

Alterations in the relative abundance of the vasoactive intestinal peptide receptors (VPAC1 and VPAC2) and functions in uterine contractility during 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.

Effect of Acrylamide Supplementation on the Population of Vasoactive Intestinal Peptide (VIP)-Like Immunoreactive Neurons in the Porcine Small Intestine.

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.

Escherichia Coli -induced Inflammation Changes the Expression of Acetylcholine Receptors (M2R, M3R, and α-7 nAChR) in the Pig Uterus.

INTRODUCTION: The influence of inflammation on the patterns of muscarinic 2 and 3 receptor subtypes (M2R and M3R), and α-7 nicotinic acetylcholine receptor (α-7 nAChR) expression in the porcine uterus was investigated. MATERIAL AND METHODS: On day three of the oestrous cycle of gilts aged 7-8 months with body weight 90-120 kg, either an E. coli suspension (E. coli group, n = 5) or saline (Sal group, n = 5) was administered into the uterine horns via laparotomy or only laparotomy was performed on control swine (Ctrl group, n = 5). After eight days, and the onset of severe acute endometritis in the E. coli group, the uterine mRNA and protein receptor expression levels were determined using real-time RT-PCR and Western blotting, respectively, with receptor localisation by immunofluorescence. RESULTS: The studied receptors were in the luminal epithelium, glands, blood vessels, and myometrial muscle cells of all gilts. The M2R mRNA level was lower in the inflamed endometrium compared to the Ctrl and Sal groups. Also in this tissue, the expression of M3R mRNA and protein was lower than in the Ctrl and Sal groups. The M3R protein level in the bacterially challenged myometrium was found to be increased compared to unadministered groups. In the endometrium of the E. coli group, the α-7 nAChR protein level was lower than in the Sal group, and in the myometrium it was reduced in relation to both the other groups. P values were ≤ 0.05 in all cases. CONCLUSION: Inflammation causes alterations in the M2R, M3R, and α-7 nAChR expression in the pig uterus, suggesting their significance in the course and repercussions of uterine inflammation.

The Influence of High and Low Doses of Acrylamide on Porcine Erythropoiesis.

INTRODUCTION: Due to the widespread occurrence of acrylamide in the environment, its likely carcinogen status, and the suitability of the pig model as a human analogue, the authors decided to evaluate the impact of high and low doses of this compound on the processes of erythropoiesis in swine bone marrow. MATERIAL AND METHODS: The experiment was carried out on Danish Landrace pigs at the age of eight weeks and body weight about 20 kg. The animals were divided into three equal groups consisting of five pigs in each. Control animals received empty gelatin capsules (placebos). Animals from the first experimental group received a low dose of acrylamide of 0.5 µg/kg b.w./day (> 99% purity; Sigma-Aldrich, Poland), and animals from the second experimental group received a dose 10 times higher. Placebos and acrylamide capsules were administered with feed every morning for 28 days. After anaesthetisation of the animals, bone marrow from the femur was collected into tubes without an anticoagulant on days 0 and 28. After drying and staining, bone marrow smears were subjected to detailed cytological evaluation using a light microscope. RESULTS: This study showed that high and low doses of acrylamide affected the process of porcine erythropoiesis. The cytotoxic effect of acrylamide on this process was demonstrated in a change of the polychromatic erythroblasts/normochromatic erythroblasts ratio. CONCLUSION: Both doses of acrylamide caused a decrease in the number of ortho- and polychromatic erythroblasts.

Inflammation changes the expression of neuropeptide Y receptors in the pig myometrium and their role in the uterine contractility.

In the experiment the influence of inflammation on neuropeptide Y (NPY) receptor subtype 1 (Y1Rs) and 2 (Y2Rs) expression pattern in the gilt myometrium and NPY effect alone or with Y1Rs and Y2Rs antagonists on contractility of an inflamed uterus were recognized. On Day 3 of the estrous cycle, either E.coli suspension (E.coli group) or saline (SAL group) were administered into uterine horns. In the control gilts (CON group), only laparotomy was carried out. Eight days later, E.coli treatment evoked severe acute endometritis, significantly reduced Y1Rs mRNA and protein expression and increased Y2Rs protein expression in myometrium in relation to the CON and SAL groups. Compared to period before NPY application, NPY (10-7 M) significantly reduced amplitude in myometrium and endometrium/myometrium and frequency in myometrium of the CON and SAL groups and amplitude in endometrium/myometrium and frequency in strips of the E.coli group. In this group, after using NPY (10-7 M), the amplitude rose significantly in both kinds of strips, while frequency fell in endometrium/myometrium in relation to the SAL group. In the CON and SAL groups, NPY (10-7 M) with Y1Rs antagonist significantly increased the amplitude in endometrium/myometrium, while with Y2Rs antagonist caused a rise in myometrium. In the E.coli group after using Y1Rs or Y2Rs antagonist and NPY (10-7 M) amplitude did not significantly change in endometrium/myometrium, and this parameter was significantly lower after using the NPY and Y1Rs antagonist than in other groups. Y1Rs antagonist and NPY (10-8, 10-7 M) significantly increased frequency in endometrium/myometrium of the CON and SAL groups. In the E.coli group, Y1Rs or Y2Rs antagonists and NPY (10-7 M) significantly increased frequency in myometrium and it was significantly higher than in the CON group. Inflammation changes Y1Rs and Y2Rs expression in myometrium of the inflamed pig uterus and NPY reduces this organ contractility by Y1Rs and Y2Rs.

Effect of Acrylamide Supplementation on the CART-, VAChT-, and nNOS-Immunoreactive Nervous Structures in the Porcine Stomach.

Acrylamide is found in food products manufactured with high-temperature processing, and exposure to acrylamide contained in food products may cause a potential risk to human health. The aim of this investigation was to demonstrate the changes in the population of CART-, nNOS-, and VAChT-immunoreactive enteric neurons in the porcine stomach in response to supplementation of low and high acrylamide doses. The study was carried out with 15 Danish landrace gilts divided into three experimental groups: the control group-animals were administered empty gelatine capsules; the low-dose group-animals were administrated a tolerable daily intake (TDI) dose (0.5 µg/kg of body weight (b.w.)/day) of acrylamide capsules, and the high-dose group-animals were administrated high-dose (ten times higher than TDI: 5 µg/kg b.w./day) acrylamide capsules for 28 days. Using the double immunofluorescence staining method, it was established that supplementation with low and high doses of acrylamide resulted in alterations of the porcine stomach neuron phenotype, which was reflected in an increased number of CART-, VAChT-, and nNOS-immunoreactive neurons. These changes were accompanied by an increased density of CART-, VAChT-, and nNOS-positive fibres. The results suggest that the enteric nervous system plays an important role in protecting the gastrointestinal tract during acrylamide intoxication.

Effect of Streptozotocin-Inducted Diabetes on the Pathophysiology of Enteric Neurons in the Small Intestine Based on the Porcine Diabetes Model.

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.

The Influence of a Hyperglycemic Condition on the Population of Somatostatin Enteric Neurons in the Porcine Gastrointestinal Tract.

Somatostatin (SOM) is the most common agent in the gastrointestinal (GI) tract that is involved in the regulation of several gastric functions, as well as in gastric disorders. Hyperglycemia, which develops as a consequence of improperly treated diabetes, can cause numerous disturbances in the appropriate functioning of the gastrointestinal tract. High glucose level is toxic to neurons. One of the lines of defense of neurons against this glucotoxicity are changes in their chemical coding. To better understood the role of SOM secreted by enteric neurons in neuronal response on elevated glucose level, pancreatic ß cells were destroyed using streptozotocin. Due to the close similarity of the pig to humans, especially the GI tract, the current study used pigs as an animal model. The results revealed that the number of enteric neurons immunoreactive to SOM (SOM-IR) in a physiological state clearly depend on the part of the GI tract studied. In turn, experimentally induced diabetes caused changes in the number of SOM-IR neurons. The least visible changes were observed in the stomach, where an increase in SOM-IR neurons was observed, only in the submucosal plexus in the corpus. However, diabetes led to an increase in the population of myenteric and submucosal neurons immunoreactive to SOM in all segments of the small intestine. The opposite situation occurred in the descending colon, where a decrease in the number of SOM-IR neurons was visible. This study underlines the significant role of SOM expressed in enteric nervous system neurons during diabetes.

Influence of Acrylamide Administration on the Neurochemical Characteristics of Enteric Nervous System (ENS) Neurons in the Porcine Duodenum.

The digestive tract, especially the small intestine, is one of the main routes of acrylamide absorption and is therefore highly exposed to the toxic effect of acrylamide contained in food. The aim of this experiment was to elucidate the effect of low (tolerable daily intake-TDI) and high (ten times higher than TDI) doses of acrylamide on the neurochemical phenotype of duodenal enteric nervous system (ENS) neurons using the pig as an animal model. The experiment was performed on 15 immature gilts of the Danish Landrace assigned to three experimental groups: control (C) group-pigs administered empty gelatine capsules, low dose (LD) group-pigs administered capsules with acrylamide at the TDI dose (0.5 µg/kg body weight (b.w.)/day), and the high dose (HD) group-pigs administered capsules with acrylamide at a ten times higher dose than the TDI (5 µg/kg b.w./day) with a morning feeding for 4 weeks. Administration of acrylamide, even in a low (TDI) dose, led to an increase in the percentage of enteric neurons immunoreactive to substance P (SP), calcitonin gene-related peptide (CGRP), galanin (GAL), neuronal nitric oxide synthase (nNOS), and vesicular acetylcholine transporter (VACHT) in the porcine duodenum. The severity of the changes clearly depended on the dose of acrylamide and the examined plexus. The obtained results suggest the participation of these neuroactive substances in acrylamide-inducted plasticity and the protection of ENS neurons, which may be an important line of defence from the harmful action of acrylamide.

The impact of low and high doses of acrylamide on the intramural neurons of the porcine ileum.

The present study was designed to assess the influence of acrylamide supplementation, in tolerable daily intake (TDI) dose and a dose ten times higher than TDI, on the neurochemical phenotype of the ENS neurons and synthesis of proinflammatory cytokines in the wall of the porcine ileum. The study was performed on 15 juvenile female Danish Landrace pigs, divided into three groups: C group- animals receiving empty gelatine capsules, LD group- animals receiving capsules with the TDI dose (0.5 µg/kg b.w./day) of acrylamide and HD group- animals receiving acrylamide in a dose ten times higher than the TDI (5 µg/kg b.w./day) in a morning meal for 28 days. It was established that supplementation of acrylamide led to an increase in substance P (SP)-, calcitonin gene-related peptide (CGRP)-, galanin (GAL)- and vesicular acetylcholine transporter (VAChT)-like immunoreactive (LI) neurons as well as a decrease in neuronal nitric oxide synthase (nNOS) -like immunoreactivity in all types of ileum intramural plexuses. Moreover, using ELISA method, an increase in the level of proinflammatory cytokines (IL-1ß, IL-6 and TNF- α) was noted in the ileum wall. The results suggest that SP, CGRP, GAL, nNOS and VACHT participate in the regulation of inflammatory conditions induced by acrylamide supplementation.