Department of Neurophysiology, Gunma University Graduate School of Medicine, Maebashi, Gunma, Japan.
PLoS One. 2012;7(11):e51015. doi: 10.1371/journal.pone.0051015. Epub 2012 Nov 30.
The murine stem cell virus (MSCV) promoter exhibits activity in mouse hematopoietic cells and embryonic stem cells. We generated transgenic mice that expressed enhanced green fluorescent protein (GFP) under the control of the MSCV promoter. We obtained 12 transgenic founder mice through 2 independent experiments and found that the bodies of 9 of the founder neonates emitted different levels of GFP fluorescence. Flow cytometric analysis of circulating leukocytes revealed that the frequency of GFP-labeled leukocytes among white blood cells ranged from 1.6% to 47.5% across the 12 transgenic mice. The bodies of 9 founder transgenic mice showed various levels of GFP expression. GFP fluorescence was consistently observed in the cerebellum, with faint or almost no fluorescence in other brain regions. In the cerebellum, 10 founders exhibited GFP expression in Purkinje cells at frequencies of 3% to 76%. Of these, 4 mice showed Purkinje cell-specific expression, while 4 and 2 mice expressed GFP in the Bergmann glia and endothelial cells, respectively. The intensity of the GFP fluorescence in the body was relative to the proportion of GFP-positive leukocytes. Moreover, the frequency of the GFP-expressing leukocytes was significantly correlated with the frequency of GFP-expressing Purkinje cells. These results suggest that the MSCV promoter is useful for preferentially expressing a transgene in Purkinje cells. In addition, the proportion of transduced leukocytes in the peripheral circulation reflects the expression level of the transgene in Purkinje cells, which can be used as a way to monitor transgene expression properties in the cerebellum without invasive techniques.
鼠干细胞病毒 (MSCV) 启动子在鼠造血细胞和胚胎干细胞中具有活性。我们生成了在 MSCV 启动子控制下表达增强型绿色荧光蛋白 (GFP) 的转基因小鼠。我们通过 2 个独立的实验获得了 12 只转基因的起始小鼠，发现 9 只新生转基因鼠的身体发出不同水平的 GFP 荧光。循环白细胞的流式细胞术分析显示，在 12 只转基因小鼠中，GFP 标记的白细胞在白细胞中的频率范围为 1.6%至 47.5%。9 只起始转基因小鼠的身体表现出不同水平的 GFP 表达。在小脑，GFP 荧光始终可见，而在其他脑区则呈现出微弱或几乎没有荧光。在小脑，10 只起始小鼠中 GFP 在浦肯野细胞中的表达频率为 3%至 76%。其中，4 只小鼠表现出浦肯野细胞特异性表达，而 4 只和 2 只小鼠分别在伯格曼胶质细胞和内皮细胞中表达 GFP。身体中的 GFP 荧光强度与 GFP 阳性白细胞的比例有关。此外，表达 GFP 的白细胞的频率与表达 GFP 的浦肯野细胞的频率显著相关。这些结果表明，MSCV 启动子可用于优先在浦肯野细胞中表达转基因。此外，外周循环中转导的白细胞的比例反映了浦肯野细胞中转基因的表达水平，可作为一种无需侵入性技术即可监测小脑中转基因表达特性的方法。
