Cell and Tissue Research

, Volume 379, Issue 1, pp 109–120 | Cite as

Quantitation and chemical coding of enteroendocrine cell populations in the human jejunum

  • Therese E Fazio Coles
  • Linda J Fothergill
  • Billie Hunne
  • Mehrdad Nikfarjam
  • Adam Testro
  • Brid Callaghan
  • Rachel M McQuade
  • John B FurnessEmail author
Regular Article


Recent studies reveal substantial species and regional differences in enteroendocrine cell (EEC) populations, including differences in patterns of hormone coexpression, which limit extrapolation between animal models and human. In this study, jejunal samples, with no histologically identifiable pathology, from patients undergoing Whipple’s procedure were investigated for the presence of gastrointestinal hormones using double- and triple-labelling immunohistochemistry and high-resolution confocal microscopy. Ten hormones (5-HT, CCK, secretin, proglucagon-derived peptides, PYY, GIP, somatostatin, neurotensin, ghrelin and motilin) were localised in EEC of the human jejunum. If only single staining is considered, the most numerous EEC were those containing 5-HT, CCK, ghrelin, GIP, motilin, secretin and proglucagon-derived peptides. All hormones had some degree of colocalisation with other hormones. This included a population of EEC in which GIP, CCK and proglucagon-derived peptides are costored, and four 5-HT cell populations, 5-HT/GIP, 5-HT/ghrelin, 5-HT/PYY, and 5-HT/secretin cell groups, and a high degree of overlap between motilin and ghrelin. The presence of 5-HT in many secretin cells is consistent across species, whereas lack of 5-HT and CCK colocalisation distinguishes human from mouse. It seems likely that the different subclasses of 5-HT cells subserve different roles. At a subcellular level, we examined the vesicular localisation of secretin and 5-HT, and found these to be separately stored. We conclude that hormone-containing cells in the human jejunum do not comply with a one-cell, one-hormone classification and that colocalisations of hormones are likely to define subtypes of EEC that have different roles.


Enteroendocrine cells Gastrointestinal hormones 5-Hydroxytryptamine Cholecystokinin Glucose-dependent insulinotropic peptide 



This work was supported by NIH (SPARC) grant ID # OT2OD023847 (PI Terry Powley) to JBF. We thank Josiane Fakhry for helpful comments on the manuscript. Confocal imaging was performed at the Biological Optical Microscopy Platform, University of Melbourne.

Funding information

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.

Informed consent

Informed consent was obtained.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. Procedures were approved by the Human Research Ethics Committee of Austin Health

Supplementary material

441_2019_3099_Fig7_ESM.png (432 kb)
Supplementary figure 1

This figure shows the counts of EEC obtained in the villi and crypts of human jejunum. The numbers are similar in the two regions, except for ghrelin and secretin, which were fewer in the crypts. c = crypts, v = villi. (PNG 432 kb)

441_2019_3099_MOESM1_ESM.tif (1.8 mb)
High resolution image (TIF 1793 kb)


  1. Adrian TE, Ferri G-L, Bacarese-Hamilton AJ, Fuessl HS, Polak JM, Bloom SR (1985) Human distribution and release of a putative’ new gut hormone, peptide YY. Gastroenterology 89:1070–1077PubMedGoogle Scholar
  2. Berger M, Gray JA, Roth BL (2009) The expanded biology of serotonin. Annu Rev Med 60:355–366PubMedPubMedCentralGoogle Scholar
  3. Buchan AMJ, Doyle AD, Accili EA (1990) Canine jejunal submucosa cultures: characterization and release of neural somatostatin. Can J Physiol Pharmacol 68:705–710PubMedGoogle Scholar
  4. Cetin Y (1990) Secretin-cells of the mammalian intestine contain serotonin. Histochemistry 93:601–606PubMedGoogle Scholar
  5. Cheung GWC, Kokorovic A, Lam CKL, Chari M, Lam TKT (2009) Intestinal cholecystokinin controls glucose production through a neuronal network. Cell Metab 10:99–109PubMedGoogle Scholar
  6. Cho H-J, Callaghan B, Bron R, Bravo DM, Furness JB (2014a) Identification of enteroendocrine cells that express TRPA1 channels in the mouse intestine. Cell Tissue Res 356:77–82PubMedGoogle Scholar
  7. Cho H-J, Robinson ES, Rivera LR, McMillan PJ, Testro A, Nikfarjam M, Bravo DM, Furness JB (2014b) Glucagon-like peptide 1 and peptide YY are in separate storage organelles in enteroendocrine cells. Cell Tissue Res 357:63–69PubMedGoogle Scholar
  8. 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
  9. Deloose E, Janssen P, Depoortere I, Tack J (2012) The migrating motor complex: control mechanisms and its role in health and disease. Nat Rev Gastroenterol Hepatol 9:271–285Google Scholar
  10. Deloose E, Vos R, Corsetti M, Depoortere I, Tack J (2015) Endogenous motilin, but not ghrelin plasma levels fluctuate in accordance with gastric phase III activity of the migrating motor complex in man. Neurogastroenterol Motil 27:63–71PubMedGoogle Scholar
  11. Diwakarla S, Fothergill LJ, Fakhry J, Callaghan B, Furness JB (2017) Heterogeneity of enterochromaffin cells within the gastrointestinal tract. Neurogastroenterol Motil 29:e13101Google Scholar
  12. Egerod KL, Engelstoft MS, Grunddal KV, Nøhr MK, Secher A, Sakata I, Pedersen J, Windeløv JA, Füchtbauer E-M, Olsen J, Sundler F, Christensen JP, Wierup N, Olsen JV, Holst JJ, Zigman JM, Poulsen SS, Schwartz TW (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. Engelstoft MS, Egerod KL, Lund ML, Schwartz TW (2013) Enteroendocrine cell types revisited. Curr Opin Pharmacol 13:912–921PubMedGoogle Scholar
  14. Fakhry J, Wang J, Martins P, Fothergill LJ, Hunne B, Prieur P, Shulkes A, Rehfeld JF, Callaghan B, Furness JB (2017) Distribution and characterisation of CCK containing enteroendocrine cells of the mouse small and large intestine. Cell Tissue Res 369:245–253PubMedGoogle Scholar
  15. 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 150:693–702PubMedPubMedCentralGoogle Scholar
  16. 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
  17. Fothergill LJ, Ringuet MT, Sioras E, Hunne B, Fazio Coles TE, Martins P, Furness JB (2019) Cellular and sub-cellular localisation of oxyntomodulin-like immunoreactivity in enteroendocrine cells of human, mouse, pig, and rat. Cell Tissue Res 375:359–369PubMedGoogle Scholar
  18. Furness JB, Hunne B, Matsuda N, Yin L, Russo D, Kato I, Fujimiya M, Patterson M, McLeod J, Andrews ZB, Bron R (2011) Investigation of the presence of ghrelin in the central nervous system of the rat and mouse. Neuroscience 193:1–9PubMedGoogle Scholar
  19. Gershon MD (2013) 5-Hydroxytryptamine (serotonin) in the gastrointestinal tract. Curr Op Endoc Diab Obes 20:14–21Google Scholar
  20. Glass LL, Calero-Nieto FJ, Jawaid W, Larraufie P, Kay RG, Göttgens B, Reimann F, Gribble FM (2017) Single-cell RNA-sequencing reveals a distinct population of proglucagon-expressing cells specific to the mouse upper small intestine. Mol Metab 6:1296–1303PubMedPubMedCentralGoogle Scholar
  21. Gribble FM, Reimann F (2016) Enteroendocrine cells: chemosensors in the intestinal epithelium. Annu Rev Physiol 78:277–299PubMedGoogle Scholar
  22. Grunddal KV, Ratner CF, Svendsen B, Sommer F, Engelstoft MS, Madsen AN, Pedersen J, Nøhr MK, Egerod KL, Nawrocki AR, Kowalski T, Howard AD, Poulsen SS, Offermanns S, Bäckhed F, Holst JJ, Holst B, Schwartz TW (2015) Neurotensin is co-expressed, co-released and acts together with GLP-1 and PYY in enteroendocrine control of metabolism. Endocrinology 157:176–194PubMedGoogle 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. He J, Irwin DM, Chen R, Zhang YP (2010) Stepwise loss of motilin and its specific receptor genes in rodents. J Mol Endocrinol 44:37–44PubMedGoogle Scholar
  25. Helander HF, Fändriks L (2012) The enteroendocrine “letter cells” – time for a new nomenclature? Scand J Gastroenterol 47:3–12PubMedGoogle Scholar
  26. Hörsch D, Fink T, Göke B, Arnold R, Büchler M, Weihe E (1994) Distribution and chemical phenotypes of neuroendocrine cells in the human anal canal. Regul Pept 54:527–542PubMedGoogle Scholar
  27. Kovacs TOG, Walsh JH, Maxwell V, Wong HC, Azuma T, Katt E (1989) Gastrin is a major mediator of the gastric phase of acid secretion in dogs: proof by monoclonal antibody neutralization. Gastroenterology 97:1406–1413PubMedGoogle Scholar
  28. 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
  29. Lo C-M, Obici S, Dong HH, Haas M, Lou D, Kim DH, Liu M, D’Alessio D, Woods SC, Tso P (2011) Impaired insulin secretion and enhanced insulin sensitivity in cholecystokinin-deficient mice. Diabetes 60:2000–2007PubMedPubMedCentralGoogle Scholar
  30. Lopez MJ, Upchurch BH, Rindi G, Leiter AB (1995) Studies in transgenic mice reveal potential relationships between secretin-producing cells and other endocrine cell types. J Biol Chem 270:885–891PubMedGoogle Scholar
  31. 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
  32. Martins P, Fakhry J, Chaves de Oliveira E, Hunne B, Fothergill LJ, Ringuet M, d’Ávila Reis D, Rehfeld JF, Callaghan B, Furness JB (2017) Analysis of enteroendocrine cell populations in the human colon. Cell Tissue Res 367:361–368Google Scholar
  33. Mawe GM, Hoffman JM (2013) Serotonin signalling in the gut—functions, dysfunctions and therapeutic targets. Nat Rev Gastroenterol Hepatol 10:473–486PubMedPubMedCentralGoogle Scholar
  34. Mortensen K, Christensen LL, Holst JJ, Orskov C (2003) GLP-1 and GIP are colocalized in a subset of endocrine cells in the small intestine. Regul Pept 114:189–196PubMedGoogle Scholar
  35. Musson MC, Jepeal LI, Finnerty JR, Wolfe MM (2011) Evolutionary expression of glucose-dependent-insulinotropic polypeptide (GIP). Regul Pept 171:26–34PubMedGoogle Scholar
  36. Patterson M, Murphy KG, le Roux CW, Ghatei MA, Bloom SR (2005) Characterization of ghrelin-like immunoreactivity in human plasma. J Clin Endocrinol Metab 90:2205–2211PubMedGoogle Scholar
  37. Peeters TL, Muls E, Janssens J, Urbain JL, Bex M, Van Cutsem E, Bouillon R (1992) Effect of motilin on gastric emptying in patients with diabetic gastroparesis. Effect of motilin on gastric emptying in patients with diabetic gastroparesis. Gastroenterology 102:97–101PubMedGoogle Scholar
  38. 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
  39. Roth KA, Gordon JI (1990) Spatial differentiation of the intestinal epithelium: analysis of enteroendocrine cells containing immunoreactive serotonin, secretin, and substance P in normal and transgenic mice. Proc Natl Acad Sci U S A 87:6408–6412PubMedPubMedCentralGoogle Scholar
  40. Säfsten B, Sjöblom M, Flemström G (2006) Serotonin increases protective duodenal bicarbonate secretion via enteric ganglia and a 5-HT4-dependent pathway. Scand J Gastroenterol 41:1279–1289PubMedGoogle Scholar
  41. Sah RP, Nagpal SJS, Mukhopadhyay D, Chari ST (2013) New insights into pancreatic cancer-induced paraneoplastic diabetes. Nat Rev Gastroenterol Hepatol 10:423–433PubMedPubMedCentralGoogle Scholar
  42. Sanger GJ, Furness JB (2016) Ghrelin and motilin receptors as drug targets for gastrointestinal disorders. Nat Rev Gastroenterol Hepatol 19:38–48Google Scholar
  43. Stengel A, Goebel M, Yakubov I, Wang L, Witcher D, Coskun T, Tache Y, Scachs G, Lambrecht NWG (2009) Identification and characterization of Nesfatin-1 immunoreactivity in endocrine cell types of the rat gastric oxyntic mucosa. Endocrinology 150:232–238PubMedGoogle Scholar
  44. Svendsen B, Pais R, Engelstoft MS, Milev NB, Richards P, Christiansen CB, Egerod KL, Jensen SM, Habib AM, Gribble FM, Schwartz TW, Reimann F, Holst JJ (2016) GLP1- and GIP-producing cells rarely overlap and differ by bombesin receptor-2 expression and responsiveness. J Endocrinol 228:39–48PubMedGoogle Scholar
  45. 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
  46. Tack J, Depoortere I, Bisschops R, Delporte C, Coulie B, Meulemans A, Janssens J, Peeters T (2006) Influence of ghrelin on interdigestive gastrointestinal motility in humans. Gut 55:327–333PubMedPubMedCentralGoogle Scholar
  47. Tatemoto K (1982) Isolation and characterization of peptide YY (PYY), a candidate gut hormone that inhibits pancreatic exocrine secretion. Proc Natl Acad Sci U S A 79:2514–2518PubMedPubMedCentralGoogle Scholar
  48. Theodorakis MJ, Carlson O, Michopoulos S, Doyle ME, Juhaszova M, Petraki K, Egan JM (2006) Human duodenal enteroendocrine cells: source of both incretin peptides, GLP-1 and GIP. Am J Physiol Endocrinol Metab 290:E550–E559PubMedPubMedCentralGoogle Scholar
  49. Tuo B-G, Isenberg JI (2003) Effect of 5-hydroxytryptamine on duodenal mucosal bicarbonate secretion in mice. Gastroenterology 125:805–814PubMedGoogle Scholar
  50. Wierup N, Björkqvist M, Westrӧm B, Pierzynowski S, Sundler F, Sjölund K (2007) Ghrelin and motilin are cosecreted from a prominent endocrine cell population in the small intestine. J Clin Endocrinol Metab 92:3573–3581PubMedGoogle Scholar
  51. Yanaihara N, Yanaihara C, Nagai K, Sato H, Shimizu F, Yamaguchi K, Abe K (1980) Motilin-like immunoreactivity in porcine, canine, human and rat tissues. Biomed Res 1:76–83Google Scholar
  52. Zhang Q, Zhu Y, Zhou W, Gao L, Yuan L, Han X (2013) Serotonin receptor 2C and insulin secretion. PLoS One 8:e54250PubMedPubMedCentralGoogle Scholar

Copyright information

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

Authors and Affiliations

  1. 1.Department of Anatomy & NeuroscienceUniversity of MelbourneParkvilleAustralia
  2. 2.Florey Institute of Neuroscience and Mental HealthParkvilleAustralia
  3. 3.Department of SurgeryUniversity of Melbourne, Austin HealthMelbourneAustralia
  4. 4.Liver and Intestinal Transplant UnitAustin HealthHeidelbergAustralia

Personalised recommendations