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.

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Alterations in Galanin-Like Immunoreactivity in the Enteric Nervous System of the Porcine Stomach Following Acrylamide Supplementation

International Journal of Molecular Sciences ◽

10.3390/ijms20133345 ◽

2019 ◽

Vol 20 (13) ◽

pp. 3345 ◽

Cited By ~ 3

Author(s):

Katarzyna Palus ◽

Krystyna Makowska ◽

Jarosław Całka

Keyword(s):

Nervous System ◽

Enteric Nervous System ◽

Daily Intake ◽

Enteric Neurons ◽

Double Labelling ◽

Recovery Processes ◽

High Doses ◽

Oxide Synthase ◽

The Impact ◽

Double Labelling Immunohistochemistry

In recent years, a significant increase in the consumption of products containing large amounts of acrylamide (e.g., chips, fries, coffee), especially among young people has been noted. The present study was created to establish the impact of acrylamide supplementation, in tolerable daily intake (TDI) dose and a dose ten times higher than TDI, on the population of galanin-like immunoreactive (GAL-LI) stomach neurons in pigs. Additionally, in the present study, the possible functional co-operation of GAL with other neuroactive substances and their role in acrylamide intoxication was investigated. Using double-labelling immunohistochemistry, alterations in the expression of GAL were examined in the porcine stomach enteric neurons after low and high doses of acrylamide supplementation. Generally, upregulation in GAL-LI immunoreactivity in both myenteric and submucous plexuses was noted in all stomach fragments studied. Additionally, the proportion of GAL-expressing cell bodies simultaneously immunoreactive to vasoactive intestinal peptide (VIP), neuronal nitric oxide synthase (nNOS) and cocaine- and amphetamine- regulated transcript peptide (CART) also increased. The results suggest neurotrophic or/and neuroprotective properties of GAL and possible co-operation of GAL with VIP, nNOS, CART in the recovery processes in the stomach enteric nervous system (ENS) neurons following acrylamide intoxication.

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Changes in Immunoreactivity of Sensory Substances within the Enteric Nervous System of the Porcine Stomach during Experimentally Induced Diabetes

Journal of Diabetes Research ◽

10.1155/2018/4735659 ◽

2018 ◽

Vol 2018 ◽

pp. 1-18 ◽

Cited By ~ 9

Author(s):

Michał Bulc ◽

Katarzyna Palus ◽

Jarosław Całka ◽

Łukasz Zielonka

Keyword(s):

Nervous System ◽

Enteric Nervous System ◽

Porcine Model ◽

Enteric Neurons ◽

Submucosal Plexus ◽

Chemical Coding ◽

Chemically Induced ◽

Calcitonin Gene ◽

Enteric Plexus ◽

Clear Increase

One of the most frequently reported disorders associated with diabetes is gastrointestinal (GI) disturbance. Although pathogenesis of these complications is multifactorial, the complicity of the enteric nervous system (ENS) in this respect has significant importance. Therefore, this paper analysed changes in substance P- (SP-), calcitonin gene-related peptide- (CGRP-), and leu5-enkephalin- (L-ENK-) like immunoreactivity (LI) in enteric stomach neurons caused by chemically induced diabetes in a porcine model. Using double immunofluorescent labelling, it was found that acute hyperglycaemia led to significant changes in the chemical coding of stomach enteric neurons. Generally, the response to artificially inducted diabetes depended on the “kind” of enteric plexus as well as the stomach region studied. A clear increase in the percentage of neurons immunoreactive to SP and CGRP was visible in the myenteric plexus (MP) in the antrum, corpus, and pylorus as well as in the submucosal plexus (SmP) in the corpus. For L-ENK, an increase in the number of L-ENK-LI neurons was observed in the MP of the antrum and SmP in the corpus, while in the MP of the corpus and pylorus, a decrease in the percentage of L-ENK-LI neurons was noted.

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

Animals ◽

10.3390/ani10040555 ◽

2020 ◽

Vol 10 (4) ◽

pp. 555 ◽

Cited By ~ 1

Author(s):

Katarzyna Palus ◽

Michał Bulc ◽

Jarosław Całka

Keyword(s):

Dose Group ◽

Low Dose ◽

Daily Intake ◽

Food Products ◽

Enteric Neurons ◽

Control Group ◽

High Dose ◽

Double Immunofluorescence ◽

High Temperature Processing ◽

High Doses

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.

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Distribution and chemical coding patterns of cocaine- and amphetamine-regulated transcript-like immunoreactive (CART-LI) neurons in the enteric nervous system of the porcine stomach cardia

Polish Journal of Veterinary Sciences ◽

10.1515/pjvs-2015-0067 ◽

2015 ◽

Vol 18 (3) ◽

pp. 515-522 ◽

Cited By ~ 6

Author(s):

W. Rękawek ◽

P. Sobiech ◽

S. Gonkowski ◽

K. Żarczyńska ◽

A. Snarska ◽

...

Keyword(s):

Nervous System ◽

Enteric Nervous System ◽

Neuronal Nitric Oxide Synthase ◽

Nerve Fibers ◽

Vesicular Acetylcholine Transporter ◽

Double Labelling ◽

Dense Network ◽

Chemical Coding ◽

Oxide Synthase ◽

Labelling Immunofluorescence

Abstract The aim of this study was to determine the presence of cocaine- and amphetamine-regulated transcript-like immunoreactive (CART-LI) neurons and co-localisation of CART with vesicular acetylcholine transporter (VAChT), neuronal nitric oxide synthase (n-NOS), vasoactive intestinal polypeptide (VIP), substance P (SP) and leu-enkephalin (LENK) in the enteric nervous system of the porcine gastric cardia by using a double-labelling immunofluorescence technique. CART-LI neurons were observed in the myenteric plexus (18.2±2.6%). A dense network of CART-LI nerve fibers was mainly observed in the muscular layer. CART showed co-localization mainly with VAChT, n-NOS, VIP and to a lesser degree with LENK and SP. Distribution of CART and its co-localization with other neurotransmitters suggest that this peptide plays an important role in gastric motility in the pig.

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Nitrergic and Substance P Immunoreactive Neurons in the Enteric Nervous System of the Bottlenose Dolphin (Tursiops truncatus) Intestine

Animals ◽

10.3390/ani11041057 ◽

2021 ◽

Vol 11 (4) ◽

pp. 1057

Author(s):

Cristiano Bombardi ◽

Anna Maria Rambaldi ◽

Giorgia Galiazzo ◽

Fiorella Giancola ◽

Jean-Marie Graïc ◽

...

Keyword(s):

Nervous System ◽

Substance P ◽

Enteric Nervous System ◽

Bottlenose Dolphin ◽

Tursiops Truncatus ◽

Bottlenose Dolphins ◽

Enteric Neurons ◽

Cetacean Species ◽

Intestinal Functions ◽

Oxide Synthase

Compared with other mammals, the digestive system of cetaceans presents some remarkable anatomical and physiological differences. However, the neurochemical features of the enteric nervous system (ENS) in these animals have only been described in part. The present study gives a description of the nitrergic and selected peptidergic systems in the myenteric plexus (MP) and submucosal plexus (SMP) of the intestine of the bottlenose dolphin (Tursiops truncatus). The distribution and morphology of neurons immunoreactive (IR) for the neuronal nitric oxide synthase (nNOS) and Substance P (SP) were immunohistochemically studied in formalin-fixed specimens from the healthy intestine of three animals, and the data were compared with those described in the literature on other mammals (human and non-human). In bottlenose dolphins, the percentages of nitrergic neurons (expressed as median and interquartile range—IQR) were 28% (IQR = 19–29) in the MP and 1% (IQR = 0–2) in the SMP, while the percentages of SP-IR neurons were 31% (IQR = 22–37) in the MP and 41% (IQR = 24–63) in the SMP. Although morphological features of nNOS- and SP-IR neurons were similar to those reported in other mammals, we found some noticeable differences in the percentages of enteric neurons. In fact, we detected a lower proportion of nNOS-IR neurons in the SMP and a higher proportion of SP-IR neurons in the MP compared to other mammals. To the best of the authors’ knowledge, this study represents the first description and quantification of nNOS-IR neurons and the first quantification of SP-IR neurons in the intestine of a cetacean species. As nNOS and SP are important mediators of intestinal functions and the nitrergic population is an important target for many neuroenteropathies, data obtained from a healthy intestine provide a necessary basis to further investigate and understand possible functional differences and motor intestinal dysfunctions/alterations in these special mammals.

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Inhibitory effect of melatonin on nitric oxide synthase in rat enteric nervous system

Gastroenterology ◽

10.1016/s0016-5085(01)80869-6 ◽

2001 ◽

Vol 120 (5) ◽

pp. A176-A176

Author(s):

P KOPPITZ ◽

M STORR ◽

D SAUR ◽

M KURJAK ◽

H ALLESCHER

Keyword(s):

Nitric Oxide ◽

Nervous System ◽

Nitric Oxide Synthase ◽

Enteric Nervous System ◽

Inhibitory Effect ◽

Oxide Synthase

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Automated computational analysis reveals structural changes in the enteric nervous system of nNOS deficient mice

Scientific Reports ◽

10.1038/s41598-021-96677-x ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Ben R. Cairns ◽

Benjamin Jevans ◽

Atchariya Chanpong ◽

Dale Moulding ◽

Conor J. McCann

Keyword(s):

Machine Learning ◽

Nervous System ◽

Enteric Nervous System ◽

Structural Changes ◽

Computational Analysis ◽

Confocal Imaging ◽

Learning Approaches ◽

Composition And Structure ◽

Oxide Synthase ◽

Computational Image Analysis

AbstractNeuronal nitric oxide synthase (nNOS) neurons play a fundamental role in inhibitory neurotransmission, within the enteric nervous system (ENS), and in the establishment of gut motility patterns. Clinically, loss or disruption of nNOS neurons has been shown in a range of enteric neuropathies. However, the effects of nNOS loss on the composition and structure of the ENS remain poorly understood. The aim of this study was to assess the structural and transcriptional consequences of loss of nNOS neurons within the murine ENS. Expression analysis demonstrated compensatory transcriptional upregulation of pan neuronal and inhibitory neuronal subtype targets within the Nos1−/− colon, compared to control C57BL/6J mice. Conventional confocal imaging; combined with novel machine learning approaches, and automated computational analysis, revealed increased interconnectivity within the Nos1−/− ENS, compared to age-matched control mice, with increases in network density, neural projections and neuronal branching. These findings provide the first direct evidence of structural and molecular remodelling of the ENS, upon loss of nNOS signalling. Further, we demonstrate the utility of machine learning approaches, and automated computational image analysis, in revealing previously undetected; yet potentially clinically relevant, changes in ENS structure which could provide improved understanding of pathological mechanisms across a host of enteric neuropathies.

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Transplantation of enteric nervous system stem cells rescues nitric oxide synthase deficient mouse colon

Nature Communications ◽

10.1038/ncomms15937 ◽

2017 ◽

Vol 8 (1) ◽

Cited By ~ 24

Author(s):

Conor J. McCann ◽

Julie E. Cooper ◽

Dipa Natarajan ◽

Benjamin Jevans ◽

Laura E. Burnett ◽

...

Keyword(s):

Nitric Oxide ◽

Stem Cells ◽

Nervous System ◽

Nitric Oxide Synthase ◽

Enteric Nervous System ◽

Deficient Mouse ◽

Mouse Colon ◽

Oxide Synthase

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In ovo transplantation of enteric nervous system precursors from vagal to sacral neural crest results in extensive hindgut colonisation

Development ◽

10.1242/dev.129.12.2785 ◽

2002 ◽

Vol 129 (12) ◽

pp. 2785-2796 ◽

Cited By ~ 1

Author(s):

Alan J. Burns ◽

Jean-Marie M. Delalande ◽

Nicole M. Le Douarin

Keyword(s):

Nervous System ◽

Neural Crest ◽

Enteric Nervous System ◽

Enteric Neurons ◽

In Ovo ◽

Neuronal Marker ◽

Sacral Region ◽

Enteric Ganglia ◽

Migration Front ◽

Sacral Neural Crest

The enteric nervous system (ENS) is derived from vagal and sacral neural crest cells (NCC). Within the embryonic avian gut, vagal NCC migrate in a rostrocaudal direction to form the majority of neurons and glia along the entire length of the gastrointestinal tract, whereas sacral NCC migrate in an opposing caudorostral direction, initially forming the nerve of Remak, and contribute a smaller number of ENS cells primarily to the distal hindgut. In this study, we have investigated the ability of vagal NCC, transplanted to the sacral region of the neuraxis, to colonise the chick hindgut and form the ENS in an experimentally generated hypoganglionic hindgut in ovo model. Results showed that when the vagal NC was transplanted into the sacral region of the neuraxis, vagal-derived ENS precursors immediately migrated away from the neural tube along characteristic pathways, with numerous cells colonising the gut mesenchyme by embryonic day (E) 4. By E7, the colorectum was extensively colonised by transplanted vagal NCC and the migration front had advanced caudorostrally to the level of the umbilicus. By E10, the stage at which sacral NCC begin to colonise the hindgut in large numbers, myenteric and submucosal plexuses in the hindgut almost entirely composed of transplanted vagal NCC, while the migration front had progressed into the pre-umbilical intestine, midway between the stomach and umbilicus. Immunohistochemical staining with the pan-neuronal marker, ANNA-1, revealed that the transplanted vagal NCC differentiated into enteric neurons, and whole-mount staining with NADPH-diaphorase showed that myenteric and submucosal ganglia formed interconnecting plexuses, similar to control animals. Furthermore, using an anti-RET antibody, widespread immunostaining was observed throughout the ENS, within a subpopulation of sacral NC-derived ENS precursors, and in the majority of transplanted vagal-to-sacral NCC. Our results demonstrate that: (1) a cell autonomous difference exists between the migration/signalling mechanisms used by sacral and vagal NCC, as transplanted vagal cells migrated along pathways normally followed by sacral cells, but did so in much larger numbers, earlier in development; (2) vagal NCC transplanted into the sacral neuraxis extensively colonised the hindgut, migrated in a caudorostral direction, differentiated into neuronal phenotypes, and formed enteric plexuses; (3) RET immunostaining occurred in vagal crest-derived ENS cells, the nerve of Remak and a subpopulation of sacral NCC within hindgut enteric ganglia.

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Abnormalities of the intestinal pacemaker cells, enteric neurons, and smooth muscle in intestinal atresia

Journal of Laboratory Physicians ◽

10.4103/jlp.jlp_94_18 ◽

2019 ◽

Vol 11 (03) ◽

pp. 180-185 ◽

Cited By ~ 1

Author(s):

Radhika krishna OH ◽

Mohammed Abdul Aleem ◽

Geetha Kayla

Keyword(s):

Nervous System ◽

Small Bowel ◽

Enteric Nervous System ◽

Gastrointestinal Motility ◽

Interstitial Cells Of Cajal ◽

Interstitial Cells ◽

Enteric Neurons ◽

Small Bowel Atresia ◽

Bowel Atresia ◽

Cells Of Cajal

Abstract BACKGROUND: Small bowel atresia is a congenital disorder that carves a substantial morbidity. Numerous postoperative gastrointestinal motility problems occur. The underlying cause of this motility disorder is still unclear. Interstitial cells of Cajal (ICC) play a major role in gastrointestinal motility. AIMS AND OBJECTIVES: To investigate the morphological changes of enteric nervous system and ICC in small bowel atresia. MATERIAL AND METHODS: Resected small bowel specimen from affected patients (n=15) were divided into three parts (proximal, distal, atretic). Standard histology and immunohistochemistry with anti C-KIT receptor antibody (CD117), calretinin and α-SMA was carried out. The density of myenteric ICCs in the proximal, atretic and distal parts was demonstrated by CD 117 while Calretinin was used for ganglion cells and nerve bundles, α-SMA highlighted muscle hypertrophy. RESULT AND CONCLUSION: The proximal and distal bowel revealed clear changes in the morphology and density of enteric nervous system and interstitial cells of Cajal..

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