Organoids as an ex vivo model for studying the serotonin system in the murine small intestine and colon epithelium

Takeshi Tsuruta, Shinichi Saito, Yosuke Osaki, Akihiro Hamada, Ayako Aoki-Yoshida, Kei Sonoyama

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Intestinal organoids were recently established as an ex vivo model of the intestinal epithelium. The present study investigated the serotonin (5-hydroxytryptamine, 5-HT) system using organoids. Organoids from murine small intestinal and colonic crypts were successfully cultured. Reverse transcription-polymerase chain reaction (RT-PCR) analysis showed that small intestinal and colonic organoids express mRNAs encoding tryptophan hydroxylase-1 (TPH1) (the rate-limiting enzyme of 5-HT synthesis), serotonin reuptake transporter (SERT), 5-HT receptor (HTR)2A, HTR2B, and HTR4. SERT mRNA levels were significantly higher in the small intestine than in the colon in both the mucosal tissues and organoids, as estimated by quantitative real-time RT-PCR. Although the 5-HT concentration and levels of chromogranin A (CgA) (an enteroendocrine cell marker), TPH1, and HTR4 mRNAs were significantly higher in the colonic mucosa than the small intestinal mucosa, they were the same in small intestinal and colonic organoids. There were no significant differences in HTR2A and HTR2B mRNA levels between the small intestine and colon in either the mucosal tissues or organoids. Immunofluorescence staining showed that the number of CgA-positive cells in the colonic organoids appeared to increase upon culturing with acetate. Acetate supplementation significantly increased CgA, TPH1, and HTR4 mRNA levels in the colonic organoids. We propose that organoids are useful for investigating the 5-HT system in the intestinal epithelium, even though colonic organoids may require gut microbiota-derived factors such as short-chain fatty acids.

Original languageEnglish
Pages (from-to)161-167
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume474
Issue number1
DOIs
Publication statusPublished - May 20 2016

Fingerprint

Organoids
Small Intestine
Serotonin
Colon
Epithelium
Tryptophan Hydroxylase
Chromogranin A
Messenger RNA
Polymerase chain reaction
Transcription
Intestinal Mucosa
Acetates
Serotonin Plasma Membrane Transport Proteins
Mucous Membrane
Volatile Fatty Acids
Reverse Transcription
Serotonin Receptors
Enteroendocrine Cells
Polymerase Chain Reaction
Cells

Keywords

  • Enterochromaffin cell
  • Intestinal organoid
  • Serotonin
  • Short-chain fatty acid

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

Cite this

Organoids as an ex vivo model for studying the serotonin system in the murine small intestine and colon epithelium. / Tsuruta, Takeshi; Saito, Shinichi; Osaki, Yosuke; Hamada, Akihiro; Aoki-Yoshida, Ayako; Sonoyama, Kei.

In: Biochemical and Biophysical Research Communications, Vol. 474, No. 1, 20.05.2016, p. 161-167.

Research output: Contribution to journalArticle

Tsuruta, Takeshi ; Saito, Shinichi ; Osaki, Yosuke ; Hamada, Akihiro ; Aoki-Yoshida, Ayako ; Sonoyama, Kei. / Organoids as an ex vivo model for studying the serotonin system in the murine small intestine and colon epithelium. In: Biochemical and Biophysical Research Communications. 2016 ; Vol. 474, No. 1. pp. 161-167.
@article{b1fc0ace031e448c81955349552f213a,
title = "Organoids as an ex vivo model for studying the serotonin system in the murine small intestine and colon epithelium",
abstract = "Intestinal organoids were recently established as an ex vivo model of the intestinal epithelium. The present study investigated the serotonin (5-hydroxytryptamine, 5-HT) system using organoids. Organoids from murine small intestinal and colonic crypts were successfully cultured. Reverse transcription-polymerase chain reaction (RT-PCR) analysis showed that small intestinal and colonic organoids express mRNAs encoding tryptophan hydroxylase-1 (TPH1) (the rate-limiting enzyme of 5-HT synthesis), serotonin reuptake transporter (SERT), 5-HT receptor (HTR)2A, HTR2B, and HTR4. SERT mRNA levels were significantly higher in the small intestine than in the colon in both the mucosal tissues and organoids, as estimated by quantitative real-time RT-PCR. Although the 5-HT concentration and levels of chromogranin A (CgA) (an enteroendocrine cell marker), TPH1, and HTR4 mRNAs were significantly higher in the colonic mucosa than the small intestinal mucosa, they were the same in small intestinal and colonic organoids. There were no significant differences in HTR2A and HTR2B mRNA levels between the small intestine and colon in either the mucosal tissues or organoids. Immunofluorescence staining showed that the number of CgA-positive cells in the colonic organoids appeared to increase upon culturing with acetate. Acetate supplementation significantly increased CgA, TPH1, and HTR4 mRNA levels in the colonic organoids. We propose that organoids are useful for investigating the 5-HT system in the intestinal epithelium, even though colonic organoids may require gut microbiota-derived factors such as short-chain fatty acids.",
keywords = "Enterochromaffin cell, Intestinal organoid, Serotonin, Short-chain fatty acid",
author = "Takeshi Tsuruta and Shinichi Saito and Yosuke Osaki and Akihiro Hamada and Ayako Aoki-Yoshida and Kei Sonoyama",
year = "2016",
month = "5",
day = "20",
doi = "10.1016/j.bbrc.2016.03.165",
language = "English",
volume = "474",
pages = "161--167",
journal = "Biochemical and Biophysical Research Communications",
issn = "0006-291X",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Organoids as an ex vivo model for studying the serotonin system in the murine small intestine and colon epithelium

AU - Tsuruta, Takeshi

AU - Saito, Shinichi

AU - Osaki, Yosuke

AU - Hamada, Akihiro

AU - Aoki-Yoshida, Ayako

AU - Sonoyama, Kei

PY - 2016/5/20

Y1 - 2016/5/20

N2 - Intestinal organoids were recently established as an ex vivo model of the intestinal epithelium. The present study investigated the serotonin (5-hydroxytryptamine, 5-HT) system using organoids. Organoids from murine small intestinal and colonic crypts were successfully cultured. Reverse transcription-polymerase chain reaction (RT-PCR) analysis showed that small intestinal and colonic organoids express mRNAs encoding tryptophan hydroxylase-1 (TPH1) (the rate-limiting enzyme of 5-HT synthesis), serotonin reuptake transporter (SERT), 5-HT receptor (HTR)2A, HTR2B, and HTR4. SERT mRNA levels were significantly higher in the small intestine than in the colon in both the mucosal tissues and organoids, as estimated by quantitative real-time RT-PCR. Although the 5-HT concentration and levels of chromogranin A (CgA) (an enteroendocrine cell marker), TPH1, and HTR4 mRNAs were significantly higher in the colonic mucosa than the small intestinal mucosa, they were the same in small intestinal and colonic organoids. There were no significant differences in HTR2A and HTR2B mRNA levels between the small intestine and colon in either the mucosal tissues or organoids. Immunofluorescence staining showed that the number of CgA-positive cells in the colonic organoids appeared to increase upon culturing with acetate. Acetate supplementation significantly increased CgA, TPH1, and HTR4 mRNA levels in the colonic organoids. We propose that organoids are useful for investigating the 5-HT system in the intestinal epithelium, even though colonic organoids may require gut microbiota-derived factors such as short-chain fatty acids.

AB - Intestinal organoids were recently established as an ex vivo model of the intestinal epithelium. The present study investigated the serotonin (5-hydroxytryptamine, 5-HT) system using organoids. Organoids from murine small intestinal and colonic crypts were successfully cultured. Reverse transcription-polymerase chain reaction (RT-PCR) analysis showed that small intestinal and colonic organoids express mRNAs encoding tryptophan hydroxylase-1 (TPH1) (the rate-limiting enzyme of 5-HT synthesis), serotonin reuptake transporter (SERT), 5-HT receptor (HTR)2A, HTR2B, and HTR4. SERT mRNA levels were significantly higher in the small intestine than in the colon in both the mucosal tissues and organoids, as estimated by quantitative real-time RT-PCR. Although the 5-HT concentration and levels of chromogranin A (CgA) (an enteroendocrine cell marker), TPH1, and HTR4 mRNAs were significantly higher in the colonic mucosa than the small intestinal mucosa, they were the same in small intestinal and colonic organoids. There were no significant differences in HTR2A and HTR2B mRNA levels between the small intestine and colon in either the mucosal tissues or organoids. Immunofluorescence staining showed that the number of CgA-positive cells in the colonic organoids appeared to increase upon culturing with acetate. Acetate supplementation significantly increased CgA, TPH1, and HTR4 mRNA levels in the colonic organoids. We propose that organoids are useful for investigating the 5-HT system in the intestinal epithelium, even though colonic organoids may require gut microbiota-derived factors such as short-chain fatty acids.

KW - Enterochromaffin cell

KW - Intestinal organoid

KW - Serotonin

KW - Short-chain fatty acid

UR - http://www.scopus.com/inward/record.url?scp=84964300259&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84964300259&partnerID=8YFLogxK

U2 - 10.1016/j.bbrc.2016.03.165

DO - 10.1016/j.bbrc.2016.03.165

M3 - Article

C2 - 27105910

AN - SCOPUS:84964300259

VL - 474

SP - 161

EP - 167

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

IS - 1

ER -