Live astrocytes visualized by green fluorescent protein in transgenic mice
Live astrocytes visualized by green fluorescent protein in transgenic mice
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DOI：
10.1006/dbio.1997.8601
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发表时间：
1997-07-01
影响因子：
2.7
通讯作者：
Messing, A
中科院分区：
文献类型：
--
作者：
Zhuo, L;Sun, B;Messing, A
绿色荧光蛋白（hGFP-S65 T）在星形胶质细胞特异性胶质细胞酸性蛋白（GFAP）启动子控制下在转基因小鼠中表达。通过北方印迹分析和共聚焦显微镜表征来自两个独立的转基因系的组织。在这两个线的表达模式是相同的，在所有组织检查，和以前发现的由相同的启动子驱动的lacZ转基因相似。使用全封片和脑切片制备物，在来自中枢神经系统多个区域的未固定或固定星形胶质细胞的细胞体和突起中观察到明亮的荧光。然而，与GFAP-lacZ转基因相反，视网膜Muller细胞响应于邻近光感受器的变性而表达GFP转基因。这些数据表明，2.2-kb的hGFAP启动子含有足够的调控元件，以指导在穆勒细胞中的表达，GFP是一个合适的报告基因，用于在哺乳动物神经系统的活制剂。这种小鼠应证明是有用的星形胶质细胞形态在发育过程中的动态变化的研究，并在响应生理和病理条件。(C)北京：科学出版社.
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 Muller 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 Muller 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. (C) 1997 Academic Press.