open access

Vol 57, No 4 (2019)
Original paper
Submitted: 2019-12-04
Accepted: 2019-12-11
Published online: 2019-12-16
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Increased expression of CART, nNOS, VIP, PACAP, SP and GAL in enteric neurons of the porcine stomach prepyloric region following hydrochloric acid infusion

Jaroslaw Calka1
DOI: 10.5603/FHC.a2019.0020
·
Pubmed: 31840794
·
Folia Histochem Cytobiol 2019;57(4):179-187.
Affiliations
  1. Department of Clinical Physiology, Faculty of Veterinary Medicine, University of Warmia and Mazury, M. Oczapowskiego 13, 10-718 Olsztyn, Poland

open access

Vol 57, No 4 (2019)
ORIGINAL PAPERS
Submitted: 2019-12-04
Accepted: 2019-12-11
Published online: 2019-12-16

Abstract

Introduction. Stomach hyperacidity leads to damage of the mucus/bicarbonate barrier, ulcerations and the development of stomach cancer. Key regulators of the mucosal barrier/luminal acid balance are neurotransmitters secreted by intramural neurons. The aim of the current study was to determine the expression of gastric neuropeptides and nNOS in the porcine stomach following hydrochloric acid instillation. We report on increased expression of enteric neurotransmitters involved in adaptive reaction to an experimentally-induced hyperacidity state.


Material and methods. The investigation was conducted on eight 12–18 kg pigs. The influence of intragastric infusion of hydrochloric acid on the expression of cocaine- and amphetamine-regulated transcript peptide (CART), neuronal nitric oxide synthase (nNOS), vasoactive intestinal polypeptide (VIP), pituitary adenylate cyclase-activating peptide (PACAP), substance P (SP) and galanin (GAL) in the submucous and myenteric gastric neurons of the pig has been studied with double immunofluorescence.

Results. A mimicked hyperacidity state significantly increased the proportion of enteric neurons immunoreactive to CART, nNOS, VIP, PACAP, SP and GAL in the submucous gastric neurons. In the myenteric plexus, a significant increase of the number of VIP-, CART- and GAL-immunoreactive (IR) neurons was found. Similarly, the percentage of myenteric nNOS-IR and PACAP-IR neurons tended to increase, while the fraction of SP-IR cells did not change.


Conclusions. Stomach hyperacidity modifies the expression of the studied neurotransmitters in a specific way depending on the location of the neurons in particular plexuses of the stomach. Increased numbers of neurons expressing CART, nNOS, VIP, PACAP, SP and GAL clearly indicate their regulatory engagement in the restoration of the physiological gastric balance following hyperacidity.

Abstract

Introduction. Stomach hyperacidity leads to damage of the mucus/bicarbonate barrier, ulcerations and the development of stomach cancer. Key regulators of the mucosal barrier/luminal acid balance are neurotransmitters secreted by intramural neurons. The aim of the current study was to determine the expression of gastric neuropeptides and nNOS in the porcine stomach following hydrochloric acid instillation. We report on increased expression of enteric neurotransmitters involved in adaptive reaction to an experimentally-induced hyperacidity state.


Material and methods. The investigation was conducted on eight 12–18 kg pigs. The influence of intragastric infusion of hydrochloric acid on the expression of cocaine- and amphetamine-regulated transcript peptide (CART), neuronal nitric oxide synthase (nNOS), vasoactive intestinal polypeptide (VIP), pituitary adenylate cyclase-activating peptide (PACAP), substance P (SP) and galanin (GAL) in the submucous and myenteric gastric neurons of the pig has been studied with double immunofluorescence.

Results. A mimicked hyperacidity state significantly increased the proportion of enteric neurons immunoreactive to CART, nNOS, VIP, PACAP, SP and GAL in the submucous gastric neurons. In the myenteric plexus, a significant increase of the number of VIP-, CART- and GAL-immunoreactive (IR) neurons was found. Similarly, the percentage of myenteric nNOS-IR and PACAP-IR neurons tended to increase, while the fraction of SP-IR cells did not change.


Conclusions. Stomach hyperacidity modifies the expression of the studied neurotransmitters in a specific way depending on the location of the neurons in particular plexuses of the stomach. Increased numbers of neurons expressing CART, nNOS, VIP, PACAP, SP and GAL clearly indicate their regulatory engagement in the restoration of the physiological gastric balance following hyperacidity.

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Keywords

pig; stomach; hyperacidity; enteric nervous system; neuropeptides; IHC

About this article
Title

Increased expression of CART, nNOS, VIP, PACAP, SP and GAL in enteric neurons of the porcine stomach prepyloric region following hydrochloric acid infusion

Journal

Folia Histochemica et Cytobiologica

Issue

Vol 57, No 4 (2019)

Article type

Original paper

Pages

179-187

Published online

2019-12-16

DOI

10.5603/FHC.a2019.0020

Pubmed

31840794

Bibliographic record

Folia Histochem Cytobiol 2019;57(4):179-187.

Keywords

pig
stomach
hyperacidity
enteric nervous system
neuropeptides
IHC

Authors

Jaroslaw Calka

References (55)
  1. Højgaard L, Mertz Nielsen A, Rune SJ. Peptic ulcer pathophysiology: acid, bicarbonate, and mucosal function. Scand J Gastroenterol Suppl. 1996; 216: 10–15.
  2. Schubert ML. Functional anatomy and physiology of gastric secretion. Curr Opin Gastroenterol. 2015; 31(6): 479–485.
  3. Alventosa-Mateu C, Ferrer-Barceló L, Huguet-Malavés JM, et al. Zollinger-Ellison syndrome. Rev Esp Enferm Dig. 2013; 105(10): 641–642.
  4. Dacha S, Razvi M, Massaad J, et al. Hypergastrinemia. Gastroenterol Rep (Oxf). 2015; 3(3): 201–208.
  5. Phan J, Benhammou JN, Pisegna JR. Gastric Hypersecretory States: Investigation and Management. Curr Treat Options Gastroenterol. 2015; 13(4): 386–397.
  6. Arafa Keshk W, Zahran SM, Katary MA, et al. Modulatory effect of silymarin on nuclear factor-erythroid-2-related factor 2 regulated redox status, nuclear factor-κB mediated inflammation and apoptosis in experimental gastric ulcer. Chem Biol Interact. 2017; 273: 266–272.
  7. Magierowski M, Magierowska K, Hubalewska-Mazgaj M, et al. Cross-talk between hydrogen sulfide and carbon monoxide in the mechanism of experimental gastric ulcers healing, regulation of gastric blood flow and accompanying inflammation. Biochem Pharmacol. 2018; 149: 131–142.
  8. Taylor S, Spugnini EP, Assaraf YG, et al. Microenvironment acidity as a major determinant of tumor chemoresistance: Proton pump inhibitors (PPIs) as a novel therapeutic approach. Drug Resist Updat. 2015; 23: 69–78.
  9. Palus K, Całka J. Neurochemical Plasticity of the Coeliac-Superior Mesenteric Ganglion Complex Neurons Projecting to the Prepyloric Area of the Porcine Stomach following Hyperacidity. Neural Plast. 2016; 2016: 8596214.
  10. Holzer P. Neural emergency system in the stomach. Gastroenterology. 1998; 114(4): 823–839.
  11. Yandrapu H, Sarosiek J. Protective Factors of the Gastric and Duodenal Mucosa: An Overview. Curr Gastroenterol Rep. 2015; 17(6): 24.
  12. Okumura T, Yamada H, Motomura W, et al. Cocaine-amphetamine-regulated transcript (CART) acts in the central nervous system to inhibit gastric acid secretion via brain corticotropin-releasing factor system. Endocrinology. 2000; 141(8): 2854–2860.
  13. Ekblad E. CART in the enteric nervous system. Peptides. 2006; 27(8): 2024–2030.
  14. Czekaj R, Majka J, Ptak-Belowska A, et al. Role of curcumin in protection of gastric mucosa against stress-induced gastric mucosal damage. Involvement of hypoacidity vasoactive mediators and sensory neuropeptides. J Physiol Pharmacol. 2016; 67:261-275. .
  15. Sepulveda B, Quispe C, Simirgiotis M, et al. Gastroprotective activity of synthetic coumarins: Role of endogenous prostaglandins, nitric oxide, non-protein sulfhydryls and vanilloid receptors. Bioorg Med Chem Lett. 2016; 26(23): 5732–5735.
  16. Phan J, Benhammou JN, Pisegna JR. Gastric Hypersecretory States: Investigation and Management. Curr Treat Options Gastroenterol. 2015; 13(4): 386–397.
  17. Zeng N, Athmann C, Kang T, et al. PACAP type I receptor activation regulates ECL cells and gastric acid secretion. J Clin Invest. 1999; 104(10): 1383–1391.
  18. Laine L, Takeuchi K, Tarnawski A. Gastric mucosal defense and cytoprotection: bench to bedside. Gastroenterology. 2008; 135(1): 41–60.
  19. Matteo F, Luca A, Rocchina C, et al. Matteo F, Luca A, Rocchina C, Marco T, Corrado B. Pathophysiology of gastric ulcer development and healing: Molecular mechanisms and novel therapeutic options. In: Chai J Ed. Peptic ulcer disease. 2011; 113-142.
  20. Norlén P, Bernsand M, Konagaya T, et al. ECL-cell histamine mobilization in conscious rats: effects of locally applied regulatory peptides, candidate neurotransmitters and inflammatory mediators. Br J Pharmacol. 2001; 134(8): 1767–1777.
  21. Makowska K, Gonkowski S, Zielonka L, et al. T2 Toxin-Induced Changes in Cocaine- and Amphetamine-Regulated Transcript (CART)-Like Immunoreactivity in the Enteric Nervous System Within Selected Fragments of the Porcine Digestive Tract. Neurotox Res. 2017; 31(1): 136–147.
  22. Krueger D, Michel K, Zeller F, et al. Neural influences on human intestinal epithelium in vitro. J Physiol. 2016; 594(2): 357–372.
  23. Burnstock G. The innervation of the gut of the brown trout salmo trutta. Q J Microsc Sci. 1959; 100: 199–220.
  24. Olsson C. Autonomic innervation of the fish gut. Acta Histochem. 2009; 111(3): 185–195.
  25. Vasina V, Barbara G, Talamonti L, et al. Enteric neuroplasticity evoked by inflammation. Auton Neurosci. 2006; 126-127: 264–272.
  26. Bulc M, Gonkowski S, Całka J. Expression of Cocaine and Amphetamine Regulated Transcript (CART) in the Porcine Intramural Neurons of Stomach in the Course of Experimentally Induced Diabetes Mellitus. J Mol Neurosci. 2015; 57(3): 376–385.
  27. Burliński PJ, Rychlik A, Całka J. Effects of inflammation and axotomy on expression of acetylcholine transferase and nitric oxide synthetase within the cocaine- and amphetamine-regulated transcript-immunoreactive neurons of the porcine descending colon. J Comp Pathol. 2014; 150(2-3): 287–296.
  28. Kaleczyc J, Klimczuk M, Franke-Radowiecka A, et al. The distribution and chemical coding of intramural neurons supplying the porcine stomach - the study on normal pigs and on animals suffering from swine dysentery. Anat Histol Embryol. 2007; 36(3): 186–193.
  29. Kasacka I, Piotrowska Z, Car H, et al. Cocaine- and amphetamine-regulated transcript : identification and distribution in human gastrointestinal tract. J Biol Regul Homeost Agents. 2012; 26(3): 419–428.
  30. Iijima K, Kanno T, Abe Y, et al. Preferential location of idiopathic peptic ulcers. Scand J Gastroenterol. 2016; 51(7): 782–787.
  31. Kanaizumi T, Nakano H, Matsui T, et al. Gastric emptying in patients with gastric and duodenal ulcer. Tohoku J Exp Med. 1989; 158(2): 133–140.
  32. Verma N, Rettenmeier AW, Schmitz-Spanke S. Recent advances in the use of Sus scrofa (pig) as a model system for proteomic studies. Proteomics. 2011; 11(4): 776–793.
  33. File SE, Pearce JB. Benzodiazepines reduce gastric ulcers induced in rats by stress. Br J Pharmacol. 1981; 74(3): 593–599.
  34. Paré W, Glavin G. Restraint stress in biomedical research: A review. Neurosci Biobehav Rev. 1986; 10(3): 339–370.
  35. Sun FP, Song YG, Cheng W, et al. Gastrin, somatostatin, G and D cells of gastric ulcer in rats. World J Gastroenterol. 2002; 8(2): 375–378.
  36. Zhou S, Yao D, Guo L, et al. Curcumin suppresses gastric cancer by inhibiting gastrin-mediated acid secretion. FEBS Open Bio. 2017; 7(8): 1078–1084.
  37. Allen A, Flemström G. Gastroduodenal mucus bicarbonate barrier: protection against acid and pepsin. Am J Physiol Cell Physiol. 2005; 288(1): C1–19.
  38. Spiess J, Villarreal J, Vale W. Isolation and sequence analysis of a somatostatin-like polypeptide from ovine hypothalamus. Biochemistry. 2002; 20(7): 1982–1988.
  39. Kemmerly T, Kaunitz JD. Gastroduodenal mucosal defense. Curr Opin Gastroenterol. 2014; 30(6): 583–588.
  40. Van Geldre LA, Lefebvre RA. Interaction of NO and VIP in gastrointestinal smooth muscle relaxation. Curr Pharm Des. 2004; 10(20): 2483–2497.
  41. Helton WS, Mulholland MM, Bunnett NW, et al. Inhibition of gastric and pancreatic secretion in dogs by CGRP: role of somatostatin. Am J Physiol. 1989; 256(4 Pt 1): G715–G720.
  42. Wallace JL, Ianaro A, de Nucci G. Gaseous Mediators in Gastrointestinal Mucosal Defense and Injury. Dig Dis Sci. 2017; 62(9): 2223–2230.
  43. Moncada S. Nitric oxide. J Hypertens Suppl. 1994; 12: S35-39. .
  44. Brown J, Hanson P, Whittle B. Nitric oxide donors increase mucus gel thickness in rat stomach. European Journal of Pharmacology. 1992; 223(1): 103–104.
  45. Walsh DA, F McWilliams D. Tachykinins and the cardiovascular system. Curr Drug Targets. 2006; 7(8): 1031–1042.
  46. Garthwaite J, Boulton CL. Nitric Oxide Signaling in the Central Nervous System. Annual Review of Physiology. 1995; 57(1): 683–706.
  47. Abdel-Salam OM, Debreceni A, Mózsik G, et al. Capsaicin-sensitive afferent sensory nerves in modulating gastric mucosal defense against noxious agents. J Physiol Paris. 1999; 93(5): 443–454.
  48. Guan X, Karpen HE, Stephens J, et al. GLP-2 receptor localizes to enteric neurons and endocrine cells expressing vasoactive peptides and mediates increased blood flow. Gastroenterology. 2006; 130(1): 150–164.
  49. Kaji I, Akiba Y, Kaunitz JD. Digestive physiology of the pig symposium: involvement of gut chemosensing in the regulation of mucosal barrier function and defense mechanisms. J Anim Sci. 2013; 91(5): 1957–1962.
  50. Wang JH, Inoue T, Higashiyama M, et al. Umami receptor activation increases duodenal bicarbonate secretion via glucagon-like peptide-2 release in rats. J Pharmacol Exp Ther. 2011; 339(2): 464–473.
  51. Forssell H, Stenquist B, Olbe L. Vagal stimulation of human gastric bicarbonate secretion. Gastroenterology. 1985; 89(3): 581–586.
  52. Takeuchi K, Yagi K, Sugamoto S, et al. Involvement of PACAP in acid-induced HCO3- response in rat duodenums. Pharmacol Res. 1998; 38(6): 475–480.
  53. Gańko M, Całka J. Prolonged acetylsalicylic-acid-supplementation-induced gastritis affects the chemical coding of the stomach innervating vagal efferent neurons in the porcine dorsal motor vagal nucleus (DMX). J Mol Neurosci. 2014; 54(2): 188–198.
  54. Goyal RK, Guo Y, Mashimo H. Advances in the physiology of gastric emptying. Neurogastroenterol Motil. 2019; 31(4): e13546.
  55. He XD, Goyal RK. Nitric oxide involvement in the peptide VIP-associated inhibitory junction potential in the guinea-pig ileum. The Journal of Physiology. 1993; 461(1): 485–499.

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