Pancreatic islet progenitor cells in Neurogenin 3-yellow fluorescent protein knock-add-on mice
Pancreatic islet progenitor cells in Neurogenin 3-yellow fluorescent protein knock-add-on mice
复制标题
DOI：
10.1210/me.2004-0243
复制
发表时间：
2004-11-01
影响因子：
--
通讯作者：
Gradwohl, G
中科院分区：
文献类型：
--
作者：
Mellitzer, G;Martín, M;Gradwohl, G
基本螺旋-环-螺旋转录因子神经原素 3 (NGN3) 控制未定型胰腺祖细胞中的内分泌细胞命运规范。 Ngn3 缺陷小鼠不会发育任何胰岛细胞，并且患有糖尿病。所有主要的胰岛细胞类型，包括产生胰岛素的 β 细胞，均源自 Ngn3 阳性内分泌祖细胞。因此，对这一未成熟细胞群的表征对于开发 1 型糖尿病细胞替代疗法的新策略特别有意义。为了进一步探索胰岛祖细胞的生物学特性，我们培育了一只小鼠，其中表达 Ngn3 的细胞使用敲入附加策略用增强型黄色荧光蛋白 (EYFP) 进行标记。在这种方法中，EYFP cDNA 被引入内分泌原转录因子 Neurogenin 3 的 3'-非翻译区，而不删除任何内源编码或调节序列。在 Ngn3(EYFP/+) 和 Ngn3(EYFP/EYFP) 小鼠中，EYFP 蛋白靶向发育中胰腺中表达 Ngn3 的祖细胞，胰岛正常发育。通过对 Ngn3(EYFP/+) 小鼠进行荧光激活细胞分选，可以从整个胚胎胰腺中纯化胰岛祖细胞，并且可以在胰腺外植体培养物中实时监测其发育。这些实验表明，在没有来自周围间充质的信号的情况下，内分泌祖细胞可以从头形成并在体外扩增，这表明内分泌定向是默认途径。 Ngn3(EYFP) 小鼠是研究正常和糖尿病动物胰岛细胞发育和新生以及确定体外生成 β 细胞的条件的宝贵工具。
The basic helix-loop-helix transcription factor Neurogenin 3 (NGN3) controls endocrine cell fate specification in uncommitted pancreatic progenitor cells. Ngn3-deficient mice do not develop any islet cells and are diabetic. All the major islet cell types, including insulin-producing beta-cells, derive from Ngn3-positive endocrine progenitor cells. Therefore, the characterization of this population of immature cells is of particular interest for the development of novel strategies for cell replacement therapies in type 1 diabetes. To explore further the biology of islet progenitor cells we have generated a mouse in which Ngn3-expressing cells are labeled with the enhanced yellow fluorescent protein (EYFP) using a knock-add-on strategy. In this approach, the EYFP cDNA is introduced into the 3'-untranslated region of the proendocrine transcription factor, Neurogenin 3, without deleting any endogenous coding or regulatory sequences. In Ngn3(EYFP/+) and Ngn3(EYFP/EYFP) mice, the EYFP protein is targeted to Ngn3-expressing progenitors in the developing pancreas, and islets develop normally. Islet progenitors can be purified from whole embryonic pancreas by fluorescence-activated cell sorting from Ngn3(EYFP/+) mice and their development can be monitored in real time in pancreas explant cultures. These experiments showed that endocrine progenitors can form de novo and expand, in vitro, in the absence of signals from the surrounding mesenchyme, suggesting that endocrine commitment is a default pathway. The Ngn3(EYFP) mice represent a valuable tool to study islet cell development and neogenesis in normal and diabetic animals as well as for the determination of the conditions to generate beta-cells in vitro.