Transgenic mice with green fluorescent protein-labeled pancreatic β-cells
Transgenic mice with green fluorescent protein-labeled pancreatic β-cells
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DOI：
10.1152/ajpendo.00321.2002
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发表时间：
2003-01-01
影响因子：
5.1
通讯作者：
Bell, GI
中科院分区：
文献类型：
--
作者：
Hara, M;Wang, XY;Bell, GI
我们已经产生了转基因小鼠，表达绿色荧光蛋白（GFP）的小鼠胰岛素I基因启动子（MIP）的控制下。MIP-GFP小鼠发育正常，在葡萄糖耐量和胰腺胰岛素含量方面与对照动物没有区别。组织学研究表明，MIP-GFP小鼠具有正常的胰岛结构，在所有胰岛的β细胞中共表达胰岛素和GFP。我们观察到GFP的表达在胰岛从胚胎天E13.5到成年。β-细胞功能的研究显示，转基因动物和对照动物胰岛之间葡萄糖诱导的细胞内钙动员没有差异。我们从MIP-GFP转基因小鼠的分离的胰岛和整个胰腺制备单细胞悬浮液，并基于它们的绿色荧光通过荧光激活细胞分选来分选β细胞。这些研究表明，新生儿（P1）胰腺中2.4 +/-0.2%（n = 6）和8周龄小鼠胰腺中0.9 +/-0.1%（n = 5）的细胞为β细胞。MIP-GFP转基因小鼠可能是研究正常和糖尿病动物β细胞生物学的有用工具。
We have generated transgenic mice that express green fluorescent protein (GFP) under the control of the mouse insulin I gene promoter (MIP). The MIP-GFP mice develop normally and are indistinguishable from control animals with respect to glucose tolerance and pancreatic insulin content. Histological studies showed that the MIP-GFP mice had normal islet architecture with coexpression of insulin and GFP in the beta-cells of all islets. We observed GFP expression in islets from embryonic day E13.5 through adulthood. Studies of beta-cell function revealed no difference in glucose-induced intracellular calcium mobilization between islets from transgenic and control animals. We prepared single-cell suspensions from both isolated islets and whole pancreas from MIP-GFP-transgenic mice and sorted the beta-cells by fluorescence-activated cell sorting based on their green fluorescence. These studies showed that 2.4 +/- 0.2% (n = 6) of the cells in the pancreas of newborn (P1) and 0.9 +/- 0.1% (n = 5) of 8-wk-old mice were beta-cells. The MIP-GFP-transgenic mouse may be a useful tool for studying beta-cell biology in normal and diabetic animals.