Prominin-1/CD133 Marks Stem Cells and Early Progenitors in Mouse Small Intestine
Prominin-1/CD133 Marks Stem Cells and Early Progenitors in Mouse Small Intestine
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DOI：
10.1053/j.gastro.2009.03.002
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发表时间：
2009-06-01
期刊：
影响因子：
29.4
通讯作者：
Clevers, Hans
中科院分区：
文献类型：
--
作者：
Snippert, Hugo J.;van Es, Johan H.;Clevers, Hans
背景和目标：Prominin-1（Prom 1）/CD 133单独或与其他细胞表面标志物组合用于从各种成体组织中鉴定和分离干细胞。我们最近发现富含亮氨酸重复序列的G蛋白偶联受体5（LGR 5）是肠干细胞的标志物，所有的胃肠上皮细胞系都来自于肠干细胞。为了确定这些标志物之间是否存在关系，我们研究了Prom 1/CD 133的肠道表达模式，并创建了敲入小鼠以观察和追踪Prom 1（+）细胞。方法：分析小肠隐窝干细胞中Prom 1 mRNA和蛋白的表达。生成Prom 1/CD 133敲入小鼠（Prom 1（-mCherry-IRIS-CreERT 2）KI），其表达红色荧光蛋白mCherry与Prom 1 C末端的融合体。敲入等位基因还含有他莫昔芬诱导型CreERT 2重组酶，允许对来自Prom 1阳性细胞的后代进行遗传追踪。结果如下：在小肠中，Prom 1 mRNA在整个隐窝的下半部分被检测到，并且不限于夹在潘氏细胞之间的罕见干细胞。Prom 1蛋白在Lgr 5（+）肠干细胞的顶膜上检测到，但也在潘氏细胞上方的过渡扩增祖细胞上检测到。对Prom 1-（mCherry-IRES-CreERT 2）KI小鼠的分析表明，Prom 1并不只在Lgr 5（+）肠干细胞中表达，而是标志着一个更大的干细胞/过渡扩增祖细胞区室。结论：Prom-1标记肠干细胞以及过境扩增祖细胞，因此它不是Lgr 5（+）肠干细胞的特异性标记物。
Background & Aims: Prominin-1(Prom1)/CD133 is used, alone or in combination with other cell surface markers, to identify and isolate stem cells from various adult tissues. We recently identified leucine-rich-repeat-containing G-protein-coupled receptor 5 (Lgr5) as a marker of the intestinal stem cells from which all cellular lineages of the gastrointestinal epithelium are derived. To determine whether there is a relationship between these markers, we investigated the intestinal expression pattern of Prom1/CD133 and created knock-in mice to visualize and trace Prom1(+) cells. Methods: We analyzed Prom1 mRNA and protein expression among stem cells within intestinal crypts. Prom1/CD133 knock-in mice (Prom1(-mCherry-IRIS-CreERT2) KI) were generated that express a fusion of red fluorescent protein mCherry with the C-terminus of Prom1. The knock-in allele also contains the tamoxifen-inducible CreERT2 recombinase, allowing for genetic tracing of progeny derived from Prom1-positive cells. Results: In the small intestine, Prom1 mRNA was detected throughout the lower half of crypts and was not restricted to the rare stem cells that are sandwiched between Paneth cells. Prom1 protein was detected at the apical membranes of Lgr5(+) intestinal stem cells, but also on the transit-amplifying progenitors located above the Paneth cells. Analyses of the Prom1-(mCherry-IRES-CreERT2) KI mice showed that Prom1 is not exclusively expressed in Lgr5(+) intestinal stem cells but marks a much larger stem cell/transit-amplifying progenitor compartment. Conclusions: Prom-1 marks intestinal stem cells, as well as transit-amplifying progenitors, so it is not a specific marker for Lgr5(+) intestinal stem cells.