Advertisement

Cell and Tissue Research

, Volume 378, Issue 3, pp 457–469 | Cite as

Distribution and co-expression patterns of specific cell markers of enteroendocrine cells in pig gastric epithelium

  • Linda J. Fothergill
  • Giorgia Galiazzo
  • Billie Hunne
  • Martin J. Stebbing
  • Josiane Fakhry
  • Frank Weissenborn
  • Therese E. Fazio Coles
  • John B. FurnessEmail author
Regular Article

Abstract

Although the pig is an accepted model species for human digestive physiology, no previous study of the pig gastric mucosa and gastric enteroendocrine cells has investigated the parallels between pig and human. In this study, we have investigated markers for each of the classes of gastric endocrine cells, gastrin, ghrelin, somatostatin, 5-hydroxytryptamine, histidine decarboxylase, and PYY cells in pig stomach. The lining of the proximal stomach consisted of a collar of stratified squamous epithelium surrounded by gastric cardiac glands in the fundus. This differs considerably from human that has only a narrow band of cardiac glands at its entrance, surrounded by a fundic mucosa consisting of oxyntic glands. However, the linings of the corpus and antrum are similar in pig and human. Likewise, the endocrine cell types are similar and similarly distributed in the two species. As in human, gastrin cells were almost exclusively in the antrum, ghrelin cells were most abundant in the oxyntic mucosa and PYY cells were rare. In the pig, 70% of enterochromaffin-like (ECL) cells in the antrum and 95% in the fundus contained 5-hydroxytryptamine (5-HT), higher proportions than in human. Unlike the enteroendocrine of the small intestine, most gastric enteroendocrine cells (EEC) did not contain colocalised hormones. This is similar to human and other species. We conclude that the pig stomach has substantial similarity to human, except that the pig has a protective lining at its entrance that may reflect the difference between a pig diet with hard abrasive components and the soft foods consumed by humans.

Keywords

Stomach Gut hormones Protective epithelium Ghrelin Enterochromaffin-like cells 

Notes

Acknowledgements

We thank Maree Cox and Jeremy Cottrell for assistance in tissue harvesting and Melinda Goga and Iain Burchill for assistance with preparation for histology. Confocal imaging was performed at the Biological Optical Microscopy Platform, University of Melbourne.

Funding

This work was financially supported by NIH (SPARC) grant ID # OT2OD023847 (PI Terry Powley) to JBF.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Ethical approval

Procedures were approved by the University of Melbourne Animal Ethics Committee (ethics approval number 1714291). All applicable National and Institutional guidelines for the care and use of animals were followed.

References

  1. Avau B, Carbone F, Tack J, Depoortere I (2013) Ghrelin signaling in the gut, its physiological properties, and therapeutic potential. Neurogastroenterol Motil 25:720–732PubMedGoogle Scholar
  2. Balasuriya GK, Hill-Yardin EL, Gershon MD, Bornstein JC (2016) A sexually dimorphic effect of cholera toxin: rapid changes in colonic motility mediated via a 5-HT3 receptor-dependent pathway in female C57Bl/6 mice. J Physiol Lond 594:4325–4338PubMedPubMedCentralGoogle Scholar
  3. Buchan AMJ, Sikora LKJ, Levy JG, McIntosh CHS, Dyck I, Brown JC (1985) An immunocytochemical investigation with monoclonal antibodies to somatostatin. Histochemistry 83:175–180PubMedGoogle Scholar
  4. Canfield SP, Spencer JE (1983) The inhibitory effects of 5-hydroxytryptamine on gastric acid secretion by the rat isolated stomach. Br J Pharmacol 78:123–129PubMedPubMedCentralGoogle Scholar
  5. Chandrasoma PT (2013) Histologic definition of gastro-esophageal reflux disease. Curr Opin Gastroenterol 29:460–467Google Scholar
  6. Cho H-J, Kosari S, Hunne B, Callaghan B, Rivera LR, Bravo DM, Furness JB (2015) Differences in hormone localisation patterns of K and L type enteroendocrine cells in the mouse and pig small intestine and colon. Cell Tissue Res 359:693–698PubMedGoogle Scholar
  7. Choi E, Roland JT, Barlow BJ, O’Neal R, Rich AE, Nam KT, Shi C, Goldenring JR (2014) Cell lineage distribution atlas of the human stomach reveals heterogeneous gland populations in the gastric antrum. Gut 63:1711–1720PubMedPubMedCentralGoogle Scholar
  8. Chuang C-N, Tanner M, Lloyd KCK, Wong H, Soll AH (1993) Endogenous somatostatin inhibits histamine release from canine gastric mucosal cells in primary culture. Am J Physiol Gastrointest Liver Physiol 28:G521–G525Google Scholar
  9. Date Y, Kojima M, Hosoda H, Sawaguchi A, Mondal MS, Suganuma T, Matsukura S, Kangawa K, Nakazato M (2000) Ghrelin, a novel growth hormone-releasing acylated peptide, is synthesized in a distinct endocrine cell type in the gastrointestinal tracts of rats and humans. Endocrinology 141:4255–4261PubMedGoogle Scholar
  10. Diwakarla S, Fothergill LJ, Fakhry J, Callaghan B, Furness JB (2017) Heterogeneity of enterochromaffin cells within the gastrointestinal tract. Neurogastroenterol Motil 29:e13101Google Scholar
  11. Dornonville De La Cour C, Björkqvist M, Sandvik AK, Bakke I, Zhao C-M, Chen D, Håkanson R (2001) A-like cells in the rat stomach contain ghrelin and do not operate under gastrin control. Regul Pept 99:141–150PubMedGoogle Scholar
  12. Egerod KL, Engelstoft MS, Grunddal KV et al (2012) A major lineage of enteroendocrine cells coexpress CCK, secretin, GIP, GLP-1, PYY, and neurotensin but not somatostatin. Endocrinology 153:5782–5795PubMedGoogle Scholar
  13. Eysselein VE, Maxwell V, Reedy T, Wünsch E, Walsh JH (1984) Similar acid stimulatory potencies of synthetic human big and little gastrins in man. J Clin Invest 73:1284–1290PubMedPubMedCentralGoogle Scholar
  14. Fakhry J, Stebbing MJ, Hunne B, Bayguinov Y, Ward SM, Sasse KC, Callaghan B, McQuade RM, Furness JB (2019) Relationships of endocrine cells to each other and to other cell types in the human gastric fundus and corpus. Cell Tissue Res 376:37–49PubMedGoogle Scholar
  15. Feldman M, Walsh JH, Wong HC (1978) Role of gastrin heptadecapeptide in the acid secretory response to amino acids in man. J Clin Invest 61:308–313PubMedPubMedCentralGoogle Scholar
  16. Fothergill LJ, Furness JB (2018) Diversity of enteroendocrine cells investigated at cellular and subcellular levels: the need for a new classification scheme. Histochem Cell Biol 150:693–702PubMedGoogle Scholar
  17. Fothergill LJ, Callaghan B, Hunne B, Bravo DM, Furness JB (2017) Costorage of enteroendocrine hormones evaluated at the cell and subcellular levels in male mice. Endocrinology 158:2113–2123PubMedGoogle Scholar
  18. Friis-Hansen L (2002) Gastric functions in gastrin gene knock-out mice. Pharmacol Toxicol 91:363–367PubMedGoogle Scholar
  19. Friis-Hansen L, Sundler F, Li Y, Gillespie PJ, Saunders TL, Greenson JK, Owyang C, Rehfeld JF, Samuelson LC (1998) Impaired gastric acid secretion in gastrin-deficient mice. Am J Physiol Gastrointest Liver Physiol 274:G561–G568Google Scholar
  20. Furness JB, Cottrell JJ, Bravo DM (2015) Comparative physiology of digestion. J Anim Sci 93:485–491PubMedGoogle Scholar
  21. Gonzalez LM, Moeser AJ, Blikslager AT (2015) Porcine models of digestive disease: the future of large animal translational research. Transl Res 166:12–27PubMedPubMedCentralGoogle Scholar
  22. Gribble FM, Reimann F, Roberts GP (2018) Gastrointestinal hormones. In: Said HM (ed) Physiology of the gastrointestinal tract, 6th edn. Academic Press, pp 31–70Google Scholar
  23. Habib AM, Richards P, Cairns LS, Rogers GJ, Bannon CAM, Parker HE, Morley TCE, Yeo GSH, Reimann F, Gribble FM (2012) Overlap of endocrine hormone expression in the mouse intestine revealed by transcriptional profiling and flow cytometry. Endocrinology 153:3054–3065PubMedPubMedCentralGoogle Scholar
  24. Håkanson R, Böttcher G, Ekblad T, Panula P, Simonsson M, Dohlsten M, Hallberg T, Sundler F (1986) Histamine in endocrine cells in the stomach. Histochemistry 86:5–17PubMedGoogle Scholar
  25. Holst JJ, Jørgensen PN, Rasmussen TN, Schmidt P (1992) Somatostatin restraint of gastrin secretion in pigs revealed by monoclonal antibody immunoneutralization. Am J Physiol Gastrointest Liver Physiol 263:G908–G912Google Scholar
  26. Hunne B, Stebbing MJ, McQuade RM, Furness JB (2019) Distributions and relationships of chemically defined enteroendocrine cells in the rat gastric mucosa. Cell Tissue Res (in press)Google Scholar
  27. Ito H, Yokozaki H, Tokumo K, Nakajo S, Tahara E (1986) Serotonin-containing EC cells in normal human gastric mucosa and in gastritis. Virchows Archiv A 409:313–323Google Scholar
  28. Kasacka I, Łebkowski W, Janiuk I, Łapińska J, Lewandowska A (2012) Immunohistochemical identification and localisation of gastrin and somatostatin in endocrine cells of human pyloric gastric mucosa. Folia Morphol (Warsz) 71:39–44Google Scholar
  29. Kim A, Park W-Y, Shin N, Lee HJ, Kim YK, Lee SJ, Hwang C-S, Park DY, Kim GH, Lee BE, Jo H-J (2015) Cardiac mucosa at the gastroesophageal junction: an Eastern perspective. World J Gastroenterol 21:9126–9133PubMedPubMedCentralGoogle Scholar
  30. Kojima M, Kangawa K (2010) Ghrelin: more than endogenous growth hormone secretagogue. Ann N Y Acad Sci 1200:140–148PubMedGoogle Scholar
  31. Kovacs TO, Lloyd KC, Lawson DC (1997) Inhibition of sham feeding-stimulated acid secretion in dogs by immunoneutralization of gastrin. Am J Physiol Gastrointest Liver Physiol 273:G399–G403Google Scholar
  32. Larsson LI, Goltermann N, De Magistris L, Rehfeld JF, Schwarz TW (1979) Somatostatin cell processes as pathways for paracrine secretion. Science 205:1393–1395PubMedGoogle Scholar
  33. Lenglinger J, See SF, Beller L, Cosentini E, Asari R, Wrba F, Riegler M, Schoppmann SF (2012) The cardia: esophageal or gastric? Critical reviewing the anatomy and histopathology of the esophagogastric junction. Acta Chir Iugosl 59:15–26PubMedGoogle Scholar
  34. Lents CA, Brown-Brandl TM, Rohrer GA, Oliver WT, Freking BA (2016) Plasma concentrations of acyl-ghrelin are associated with average daily gain and feeding behavior in grow-finish pigs. Domest Anim Endocrinol 55:107–113PubMedGoogle Scholar
  35. LePard KJ, Chi J, Mohammed JR, Gidener S, Stephens RL Jr (1996) Gastric antisecretory effect of serotonin: quantitation of release and site of action. Am J Physiol Endocrinol Metab 271:E669–E677Google Scholar
  36. Levin F, Edholm T, Schmidt PT, Grybäck P, Jacobsson H, Degerblad M, Höybye C, Holst JJ, Rehfeld JF, Hellström PM, Näslund E (2006) Ghrelin stimulates gastric emptying and hunger in normal-weight humans. J Clin Endocrinol Metab 91:3296–3302PubMedGoogle Scholar
  37. Li Y-Y (2003) Mechanisms for regulation of gastrin and somatostatin release from isolated rat stomach during gastric distention. World J Gastroenterol 9:129–133PubMedPubMedCentralGoogle Scholar
  38. Martin AM, Young RL, Leong L, Rogers GB, Spencer NJ, Jessup CF, Keating DJ (2017) The diverse metabolic roles of peripheral serotonin. Endocrinology 158:1049–1063PubMedGoogle Scholar
  39. Mawe GM, Hoffman JM (2013) Serotonin signalling in the gut—functions, dysfunctions and therapeutic targets. Nat Rev Gastroenterol Hepatol 10:473–486PubMedPubMedCentralGoogle Scholar
  40. Meulengracht E (1935) The glands of the stomach in relation to pernicious anaemia; with special reference to the glands in the pyloric region. Proc R Soc Med 28:841–870PubMedPubMedCentralGoogle Scholar
  41. Mizutani M, Atsuchi K, Asakawa A, Matsuda N, Fujimura M, Inui A, Kato I, Fujimiya M (2009) Localization of acyl ghrelin- and des-acyl ghrelin-immunoreactive cells in the rat stomach and their responses to intragastric pH. Am J Physiol Gastrointest Liver Physiol 297:G974–G980PubMedGoogle Scholar
  42. Payne SC, Furness JB, Stebbing MJ (2018) Bioelectric neuromodulation for gastrointestinal disorders: effectiveness and mechanisms. Nat Rev Gastroenterol Hepatol 16:89–105Google Scholar
  43. Rehfeld JF, Friis-Hansen L, Goetze JP, Hansen TVO (2007) The biology of cholecystokinin and gastrin peptides. Curr Top Med Chem 7:1154–1165PubMedGoogle Scholar
  44. Reynaud Y, Fakhry J, Fothergill L, Callaghan B, Ringuet MT, Hunne B, Bravo DM, Furness JB (2016) The chemical coding of 5-hydroxytryptamine containing enteroendocrine cells in the mouse gastrointestinal tract. Cell Tissue Res 364:489–497PubMedGoogle Scholar
  45. Rindi G, Necchi V, Savio A, Torsello A, Zoli M, Locatelli V, Raimondo F, Cocchi D, Solcia E (2002) Characterisation of gastric ghrelin cells in man and other mammals: studies in adult and fetal tissues. Histochem Cell Biol 117:511–519PubMedGoogle Scholar
  46. Roura E, Koopmans S-J, Lallès J-P, Le Huerou-Luron I, de Jager N, Schuurman T, Val-Laillet D (2016) Critical review evaluating the pig as a model for human nutritional physiology. Nutr Res Rev 29:60–90PubMedGoogle Scholar
  47. Salfen BE, Carroll JA, Keisler DH, Strauch TA (2004) Effects of exogenous ghrelin on feed intake, weight gain, behavior, and endocrine responses in weanling pigs. J Anim Sci 82:1957–1966PubMedGoogle Scholar
  48. Sandvik AK, Dimaline R, Mårvik R, Brenna E, Waldum HL (1994) Gastrin regulates histidine decarboxylase activity and mRNA abundance in rat oxyntic mucosa. Am J Physiol Gastrointest Liver Physiol 267:G254–G258Google Scholar
  49. Schubert ML, Peura DA (2008) Control of gastric acid secretion in health and disease. Gastroenterology 134:1842–1860PubMedGoogle Scholar
  50. Schubert ML, Edwards NF, Makhlouf GM (1988) Regulation of gastric somatostatin secretion in the mouse by luminal acidity: a local feedback mechanism. Gastroenterology 94:317–322PubMedGoogle Scholar
  51. Smolka AJ, Larsen KA, Hammond CE (2000) Location of a cytoplasmic epitope for monoclonal antibody HK 12.18 on H,K-ATPase α subunit. Biochem Biophys Res Commun 273:942–947PubMedGoogle Scholar
  52. Soll AH, Walsh JH (1979) Regulation of gastric acid secretion. Annu Rev Physiol 41:35–53PubMedGoogle Scholar
  53. Sykaras AG, Demenis C, Cheng L, Pisitkun T, Mclaughlin JT, Fenton RA, Smith CP (2014) Duodenal CCK cells from male mice express multiple hormones including ghrelin. Endocrinology 155:3339–3351PubMedGoogle Scholar
  54. Szelenyi I, Herold H, Göthert M (1994) Emesis induced in domestic pigs: a new experimental tool for detection of antiemetic drugs and for evaluation of emetogenic potential of new anticancer agents. J Pharmacol Toxicol Methods 32:109–116PubMedGoogle Scholar
  55. Vitari F, Di Giancamillo A, Deponti D, Carollo V, Domeneghini C (2012) Distribution of ghrelin-producing cells in the gastrointestinal tract of pigs at different ages. Vet Res Commun 36:71–80PubMedGoogle Scholar
  56. Vuyyuru L, Schubert ML, Harrington L, Arimura A, Makhlouf GM (1995) Dual inhibitory pathways link antral somatostatin and histamine secretion in human, dog, and rat stomach. Gastroenterology 109:1566–1574PubMedGoogle Scholar
  57. Vuyyuru L, Harrington L, Arimura A, Schubert ML (1997) Reciprocal inhibitory paracrine pathways link histamine and somatostatin secretion in the fundus of the stomach. Am J Physiol Gastrointest Liver Physiol 273:G106–G111Google Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Linda J. Fothergill
    • 1
    • 2
  • Giorgia Galiazzo
    • 1
  • Billie Hunne
    • 1
  • Martin J. Stebbing
    • 1
    • 2
  • Josiane Fakhry
    • 1
  • Frank Weissenborn
    • 3
  • Therese E. Fazio Coles
    • 1
  • John B. Furness
    • 1
    • 2
    • 3
    Email author
  1. 1.Department of Anatomy & NeuroscienceUniversity of MelbourneParkvilleAustralia
  2. 2.Florey Institute of Neuroscience and Mental HealthParkvilleAustralia
  3. 3.Department of Agriculture and FoodUniversity of MelbourneParkvilleAustralia

Personalised recommendations