A Major Lineage of Enteroendocrine Cells Coexpress CCK, Secretin, GIP, GLP-1, PYY, and Neurotensin but Not Somatostatin
A Major Lineage of Enteroendocrine Cells Coexpress CCK, Secretin, GIP, GLP-1, PYY, and Neurotensin but Not Somatostatin
复制标题
DOI：
10.1210/en.2012-1595
复制
发表时间：
2012-12-01
期刊：
影响因子：
4.8
通讯作者：
Schwartz, Thue W.
中科院分区：
文献类型：
--
作者：
Egerod, Kristoffer L.;Engelstoft, Maja S.;Schwartz, Thue W.
肠内分泌细胞，如十二指肠胆囊收缩素（CCK细胞）通常被认为局限于胃肠道的某些部分，并且仅储存和释放来源于单一肽前体的肽。然而，在目前的研究中，在CCK启动子控制下表达增强绿色荧光蛋白（eGFP）的转基因小鼠显示出CCK-eGFP阳性细胞在整个肠道中的分布模式。分离的、facs纯化的CCK- egfp阳性细胞的定量PCR和液相色谱-质谱蛋白质组学分析表明，不仅表达CCK，还表达胰高血糖素样肽1 （GLP-1）、胃抑制肽（GIP）、YY肽（PYY）、神经紧张素和分泌素，但不表达生长抑素。免疫组织化学证实了这种表达模式。通过分离细胞群的免疫组织化学和FACS分析，在隐窝和绒毛中均观察到广泛的共表达现象。单细胞定量PCR表明，大约一半的十二指肠CCK- egfp细胞除了CCK外还表达一种肽前体，而另外一小部分除了CCK外还表达两种肽前体。通过胰高血糖素前启动子控制下的人白喉毒素受体表达的细胞消融研究进一步证实了这种共表达模式，其中受体的激活不仅导致GLP-1细胞显著减少，而且导致PYY、神经紧张素、GIP、CCK和分泌素细胞显著减少，而生长抑素细胞则未受影响。通过免疫组织化学双染色在人小肠中证实了共表达模式的关键元件。结论是，成熟肠内分泌细胞谱系具有共表达一组功能相关肽的能力：CCK，分泌素，GIP, GLP-1， PYY和神经紧张素，提示治疗和预防糖尿病和肥胖的潜在治疗靶点。（内分泌学153:5782-5795,2012）
Enteroendocrine cells such as duodenal cholecystokinin (CCK cells) are generally thought to be confined to certain segments of the gastrointestinal (GI) tract and to store and release peptides derived from only a single peptide precursor. In the current study, however, transgenic mice expressing enhanced green fluorescent protein (eGFP) under the control of the CCK promoter demonstrated a distribution pattern of CCK-eGFP positive cells that extended throughout the intestine. Quantitative PCR and liquid chromatography-mass spectrometry proteomic analyses of isolated, FACS-purified CCK-eGFP-positive cells demonstrated expression of not only CCK but also glucagon-like peptide 1 (GLP-1), gastric inhibitory peptide (GIP), peptide YY (PYY), neurotensin, and secretin, but not somatostatin. Immunohistochemistry confirmed this expression pattern. The broad coexpression phenomenon was observed both in crypts and villi as demonstrated by immunohistochemistry and FACS analysis of separated cell populations. Single-cell quantitative PCR indicated that approximately half of the duodenal CCK-eGFP cells express one peptide precursor in addition to CCK, whereas an additional smaller fraction expresses two peptide precursors in addition to CCK. The coexpression pattern was further confirmed through a cell ablation study based on expression of the human diphtheria toxin receptor under the control of the proglucagon promoter, in which activation of the receptor resulted in a marked reduction not only in GLP-1 cells, but also PYY, neurotensin, GIP, CCK, and secretin cells, whereas somatostatin cells were spared. Key elements of the coexpression pattern were confirmed by immunohistochemical double staining in human small intestine. It is concluded that a lineage of mature enteroendocrine cells have the ability to coexpress members of a group of functionally related peptides: CCK, secretin, GIP, GLP-1, PYY, and neurotensin, suggesting a potential therapeutic target for the treatment and prevention of diabetes and obesity. (Endocrinology 153: 5782-5795, 2012)