Zhuo L, Sun B, Zhang C L, Fine A, Chiu S Y, Messing A
Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, 53706, USA.
Dev Biol. 1997 Jul 1;187(1):36-42. doi: 10.1006/dbio.1997.8601.
Green fluorescent protein (hGFP-S65T) was expressed in transgenic mice under the control of the astrocyte-specific glial fibrillary acidic protein (GFAP) promoter. Tissues from two independent transgenic lines were characterized by Northern blot analysis and by confocal microscopy. The expression pattern in these two lines was identical in all tissues examined, and similar to that found previously with a lacZ transgene driven by the same promoter. Bright fluorescence was observed in the cell bodies and processes of unfixed or fixed astrocytes, using both whole mount and brain slice preparations, from multiple areas of the central nervous system. However, in contrast to GFAP-lacZ transgenics, retinal Müller cells expressed the GFP transgene in response to degeneration of neighboring photoreceptors. These data indicate that the 2.2-kb hGFAP promoter contains sufficient regulatory elements to direct expression in Müller cells, and that GFP is a suitable reporter gene for use in living preparations of the mammalian nervous system. Such mice should prove useful for studies of dynamic changes in astrocyte morphology during development, and in response to physiological and pathological conditions.
绿色荧光蛋白（hGFP-S65T）在星形胶质细胞特异性胶质纤维酸性蛋白（GFAP）启动子的控制下在转基因小鼠中表达。通过Northern印迹分析和共聚焦显微镜对来自两个独立转基因系的组织进行了表征。在所有检测的组织中，这两个系的表达模式是相同的，并且与先前由相同启动子驱动的lacZ转基因所发现的模式相似。使用整装标本和脑切片标本，在中枢神经系统的多个区域，未固定或固定的星形胶质细胞的细胞体和突起中均观察到明亮的荧光。然而，与GFAP-lacZ转基因小鼠不同，视网膜穆勒细胞在邻近光感受器退化时表达GFP转基因。这些数据表明，2.2 kb的hGFAP启动子包含足够的调控元件来指导在穆勒细胞中的表达，并且GFP是用于哺乳动物神经系统活体标本的合适报告基因。此类小鼠应被证明对研究发育过程中以及对生理和病理条件作出反应时星形胶质细胞形态的动态变化有用。
