Stimulation of motilin secretion by bile, free fatty acids, and acidification in human duodenal organoids

Objective: Motilin is a proximal small intestinal hormone with roles in gastrointestinal motility, gallbladder emptying, and hunger initiation. In vivo motilin release is stimulated by fats, bile, and duodenal acidification but the underlying molecular mechanisms of motilin secretion remain poorly u...

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Autores principales:	Emily L. Miedzybrodzka, Rachel E. Foreman, Van B. Lu, Amy L. George, Christopher A. Smith, Pierre Larraufie, Richard G. Kay, Deborah A. Goldspink, Frank Reimann, Fiona M. Gribble
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	Motilin Secretion Human intestinal organoids Enteroendocrine hormones Internal medicine RC31-1245
Acceso en línea:	https://doaj.org/article/7038fb1fad7e44e6bafe941133cf6234
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spelling	oai:doaj.org-article:7038fb1fad7e44e6bafe941133cf62342021-11-10T04:25:29ZStimulation of motilin secretion by bile, free fatty acids, and acidification in human duodenal organoids2212-877810.1016/j.molmet.2021.101356https://doaj.org/article/7038fb1fad7e44e6bafe941133cf62342021-12-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2212877821002039https://doaj.org/toc/2212-8778Objective: Motilin is a proximal small intestinal hormone with roles in gastrointestinal motility, gallbladder emptying, and hunger initiation. In vivo motilin release is stimulated by fats, bile, and duodenal acidification but the underlying molecular mechanisms of motilin secretion remain poorly understood. This study aimed to establish the key signaling pathways involved in the regulation of secretion from human motilin-expressing M-cells. Methods: Human duodenal organoids were CRISPR-Cas9 modified to express the fluorescent protein Venus or the Ca2+ sensor GCaMP7s under control of the endogenous motilin promoter. This enabled the identification and purification of M-cells for bulk RNA sequencing, peptidomics, calcium imaging, and electrophysiology. Motilin secretion from 2D organoid-derived cultures was measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS), in parallel with other gut hormones. Results: Human duodenal M-cells synthesize active forms of motilin and acyl-ghrelin in organoid culture, and also co-express cholecystokinin (CCK). Activation of the bile acid receptor GPBAR1 stimulated a 3.4-fold increase in motilin secretion and increased action potential firing. Agonists of the long-chain fatty acid receptor FFA1 and monoacylglycerol receptor GPR119 stimulated secretion by 2.4-fold and 1.5-fold, respectively. Acidification (pH 5.0) was a potent stimulus of M-cell calcium elevation and electrical activity, an effect attributable to acid-sensing ion channels, and a modest inducer of motilin release. Conclusions: This study presents the first in-depth transcriptomic and functional characterization of human duodenal motilin-expressing cells. We identify several receptors important for the postprandial and interdigestive regulation of motilin release.Emily L. MiedzybrodzkaRachel E. ForemanVan B. LuAmy L. GeorgeChristopher A. SmithPierre LarraufieRichard G. KayDeborah A. GoldspinkFrank ReimannFiona M. GribbleElsevierarticleMotilinSecretionHuman intestinal organoidsEnteroendocrine hormonesInternal medicineRC31-1245ENMolecular Metabolism, Vol 54, Iss , Pp 101356- (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Motilin Secretion Human intestinal organoids Enteroendocrine hormones Internal medicine RC31-1245
spellingShingle	Motilin Secretion Human intestinal organoids Enteroendocrine hormones Internal medicine RC31-1245 Emily L. Miedzybrodzka Rachel E. Foreman Van B. Lu Amy L. George Christopher A. Smith Pierre Larraufie Richard G. Kay Deborah A. Goldspink Frank Reimann Fiona M. Gribble Stimulation of motilin secretion by bile, free fatty acids, and acidification in human duodenal organoids
description	Objective: Motilin is a proximal small intestinal hormone with roles in gastrointestinal motility, gallbladder emptying, and hunger initiation. In vivo motilin release is stimulated by fats, bile, and duodenal acidification but the underlying molecular mechanisms of motilin secretion remain poorly understood. This study aimed to establish the key signaling pathways involved in the regulation of secretion from human motilin-expressing M-cells. Methods: Human duodenal organoids were CRISPR-Cas9 modified to express the fluorescent protein Venus or the Ca2+ sensor GCaMP7s under control of the endogenous motilin promoter. This enabled the identification and purification of M-cells for bulk RNA sequencing, peptidomics, calcium imaging, and electrophysiology. Motilin secretion from 2D organoid-derived cultures was measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS), in parallel with other gut hormones. Results: Human duodenal M-cells synthesize active forms of motilin and acyl-ghrelin in organoid culture, and also co-express cholecystokinin (CCK). Activation of the bile acid receptor GPBAR1 stimulated a 3.4-fold increase in motilin secretion and increased action potential firing. Agonists of the long-chain fatty acid receptor FFA1 and monoacylglycerol receptor GPR119 stimulated secretion by 2.4-fold and 1.5-fold, respectively. Acidification (pH 5.0) was a potent stimulus of M-cell calcium elevation and electrical activity, an effect attributable to acid-sensing ion channels, and a modest inducer of motilin release. Conclusions: This study presents the first in-depth transcriptomic and functional characterization of human duodenal motilin-expressing cells. We identify several receptors important for the postprandial and interdigestive regulation of motilin release.
format	article
author	Emily L. Miedzybrodzka Rachel E. Foreman Van B. Lu Amy L. George Christopher A. Smith Pierre Larraufie Richard G. Kay Deborah A. Goldspink Frank Reimann Fiona M. Gribble
author_facet	Emily L. Miedzybrodzka Rachel E. Foreman Van B. Lu Amy L. George Christopher A. Smith Pierre Larraufie Richard G. Kay Deborah A. Goldspink Frank Reimann Fiona M. Gribble
author_sort	Emily L. Miedzybrodzka
title	Stimulation of motilin secretion by bile, free fatty acids, and acidification in human duodenal organoids
title_short	Stimulation of motilin secretion by bile, free fatty acids, and acidification in human duodenal organoids
title_full	Stimulation of motilin secretion by bile, free fatty acids, and acidification in human duodenal organoids
title_fullStr	Stimulation of motilin secretion by bile, free fatty acids, and acidification in human duodenal organoids
title_full_unstemmed	Stimulation of motilin secretion by bile, free fatty acids, and acidification in human duodenal organoids
title_sort	stimulation of motilin secretion by bile, free fatty acids, and acidification in human duodenal organoids
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/7038fb1fad7e44e6bafe941133cf6234
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Stimulation of motilin secretion by bile, free fatty acids, and acidification in human duodenal organoids

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