Organoids as an ex vivo model for studying the serotonin system in the murine small intestine and colon epithelium.
Organoids as an ex vivo model for studying the serotonin system in the murine small intestine and colon epithelium.
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DOI：
10.1016/j.bbrc.2016.03.165
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发表时间：
2016-05
影响因子：
3.1
通讯作者：
Takeshi Tsuruta;S. Saito;Y. Osaki;Akihiro Hamada;A. Aoki-Yoshida;K. Sonoyama
中科院分区：
文献类型：
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作者：
Takeshi Tsuruta;S. Saito;Y. Osaki;Akihiro Hamada;A. Aoki-Yoshida;K. Sonoyama
肠类器官最近被建立为肠上皮的活体模型。本研究用有机化合物研究了5-羟色胺(5-羟色胺，5-羟色胺)系统。成功地培养了小鼠小肠和结肠隐窝中的有机化合物。逆转录-聚合酶链式反应(RT-PCR)分析表明，小肠和结肠类器官表达编码5-羟色胺合成的限速酶色氨酸羟基酶1(TPH1)、5-羟色胺再摄取转运体(SERT)、5-羟色胺受体(HTR)2A、HTR2B和HTR4的mRNAs。实时荧光定量RT-PCR检测结果显示，无论是粘膜组织还是器官，小肠中SERT基因的表达水平均显著高于结肠。尽管5-羟色胺(5-羟色胺)浓度和肠道内分泌细胞标志物嗜铬粒蛋白A(CGA)、TPH1和HTR4的mRNAs水平在结肠粘膜显著高于小肠粘膜，但在小肠和结肠器官中的浓度和水平是相同的。小肠和结肠组织中HTR2A和HTR2B的表达水平在粘膜组织和器官中均无显著差异。免疫荧光染色显示，加入醋酸盐培养后，结肠细胞器中CgA阳性细胞数明显增加。添加醋酸盐显著提高了结肠器官中CGA、TPH1和HTR4的mRNA水平。我们认为有机化合物对于研究肠道上皮细胞中的5-羟色胺系统是有用的，尽管结肠有机化合物可能需要肠道微生物群衍生的因子，如短链脂肪酸。
Intestinal organoids were recently established as anex vivomodel 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.