Department of Surgery, UCSD Moores Cancer Center, La Jolla, CA 92093-0987, USA.
Pancreas. 2013 Aug;42(6):952-8. doi: 10.1097/MPA.0b013e31828643df.
To develop a mouse model for multispectral fluorescence imaging of the pancreas and pancreatic microenvironment.
Cre/loxP technology was used to develop this model. We crossed mT/mG indicator mice, engineered to constitutively express a conditional tdTomato transgene that converts to green fluorescent protein (GFP) expression after exposure to Cre recombinase, with Pdx1-Cre transgenic mice. To characterize this model for studies of pancreas biology, we performed bright light and fluorescence imaging of body cavities and intact organs and confocal microscopy of pancreata from offspring of Pdx1-Cre and mT/mG crosses.
Pdx1-Cre-mT/mG mice demonstrated bright GFP expression within the pancreas and duodenum and intense tdTomato expression in all other organs. Green fluorescent protein expression was mosaic in Pdx1-Cre-mT/mG pancreata, with most showing extensive conversion from tdTomato to GFP expression within the epithelial-derived elements of the pancreatic parenchyma. Because both GFP and tdTomato are membrane targeted, individual cell borders were clearly outlined in confocal images of mT/mG pancreata.
This mouse model enables multispectral fluorescence imaging of individual cells and cell processes at the microscopic level of the pancreatic microenvironment; it should prove valuable for a variety of fluorescence imaging studies, ranging from pancreatic development to pancreatic cancer biology.
开发一种用于胰腺和胰腺微环境多光谱荧光成像的小鼠模型。
利用 Cre/loxP 技术构建该模型。我们将持续表达条件性 tdTomato 转基因的 mT/mG 报告小鼠(该转基因在暴露于 Cre 重组酶后会转化为绿色荧光蛋白 [GFP] 表达)与 Pdx1-Cre 转基因小鼠进行杂交。为了对该模型进行胰腺生物学研究,我们对 Pdx1-Cre 和 mT/mG 杂交后代的体腔和完整器官进行了明场和荧光成像,并对胰腺进行了共聚焦显微镜检查。
Pdx1-Cre-mT/mG 小鼠的胰腺和十二指肠内可见明亮的 GFP 表达,所有其他器官均可见强烈的 tdTomato 表达。Pdx1-Cre-mT/mG 胰腺内 GFP 表达呈镶嵌状,大多数上皮源性胰腺实质细胞内的 tdTomato 到 GFP 的转化广泛存在。由于 GFP 和 tdTomato 均定位于细胞膜,因此 mT/mG 胰腺的共聚焦图像中可以清晰地勾勒出单个细胞边界。
该小鼠模型可实现胰腺微环境中单个细胞和细胞过程的多光谱荧光成像;它应能为从胰腺发育到胰腺癌生物学的各种荧光成像研究提供有力支持。