利用谱系追踪方法探究Lgr5在胰腺组织及其类器官中的表达
收稿日期: 2022-01-26
修回日期: 2022-03-26
网络出版日期: 2022-04-18
基金资助
国家自然科学基金面上项目资助(32072801)
Study on Lgr5 expression in pancreas tissues and organoids by lineage tracing
Received date: 2022-01-26
Revised date: 2022-03-26
Online published: 2022-04-18
Supported by
Supported by the National Natural Science Foundation of China(32072801)
富含亮氨酸重复序列G蛋白偶联受体5 (leucine-rich repeat containing G protein-coupled receptor 5, Lgr5)在体内分布广泛,可以作为多种上皮组织(包括小肠、结肠、胃和毛囊)中干细胞的标记物。为了探究小鼠(Mus musculus)胰腺发育过程中导管上皮细胞及体外培养的胰腺导管类器官中Lgr5的表达情况,本研究利用Lgr5-CreERT2+/-和Rosa26-mTmG杂交后的转基因小鼠,经Tamoxifen (他莫昔芬)诱导后,观察不同发育阶段胰腺组织切片的荧光表达情况,并通过三维培养建立成体小鼠胰腺导管类器官,观察诱导后类器官细胞中的荧光变化。结果显示：Tamoxifen诱导的正常成体转基因小鼠胰腺导管内未检测到表达Lgr5的细胞;通过对孕鼠及哺乳母鼠注射Tamoxifen,在胚胎发育15.5 d和新生小鼠胰腺中也未发现Lgr5阳性细胞;但是将4-hydroxy- Tamoxifen (4-羟基-他莫昔芬)添加到培养基中,在Lgr5-CreERT2+/-;Rosa26-mTmG转基因小鼠胰腺导管来源的类器官中检测到部分细胞表达Lgr5。本研究结果证实,在成体及胚胎胰腺组织中没有检测到Lgr5表达,但在体外培养的胰腺导管类器官细胞中检测到Lgr5表达。本研究为探索胰腺发育过程中干/祖细胞特异性表达基因奠定了基础。
闫炳儒, 艾显辉, 刘淼, 田丽红, 梁洋, 滕春波 . 利用谱系追踪方法探究Lgr5在胰腺组织及其类器官中的表达[J]. 遗传, 2022 , 44(5) : 432 -441 . DOI: 10.16288/j.yczz.22-022
Leucine rich repeat containing G protein-coupled receptor 5(Lgr5) is widely expressed in multiple tissues and can be used as a stem cell marker in a variety of epithelial organs (including the small intestine, colon, stomach and hair follicles). In this study, we used Lgr5-CreERT2+/- and Rosa26-mTmG hybridized transgenic mice to investigate the expression of Lgr5 in both ductal epithelial cells during pancreas development and in vitro cultured pancreatic duct organoids. After induction with Tamoxifen, the Lgr5 expression was analyzed by detecting the enhanced green fluorescence protein in the pancreatic tissue sections in adult animals and embryos at different developmental stages. The results showed that Lgr5 expression was detected neither in adult pancreatic duct epithelia nor in the embryonic pancreatic tissues at day 15.5 or in newborn mice. However, when 4-hydroxy-Tamoxifen was supplemented to the culture medium, EGFP could be detected in the primary pancreatic duct organoids from Lgr5-Cre ERT2+/-; Rosa26-mTmG mice. These results suggested that Lgr5 was not expressed in adult and embryonic pancreatic tissues; but could be expressed in the cultured pancreas ductal organoids. The research lays the foundation for exploring specific gene expression patterns in stem/progenitor cells during pancreatic development.
Key words： pancreas; duct; Lgr5; organoid; lineage tracing
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