Dual transgenic reporter mice as a tool for monitoring expression of glial fibrillary acidic protein.
Dual transgenic reporter mice as a tool for monitoring expression of glial fibrillary acidic protein.
复制标题
DOI：
10.1111/j.1471-4159.2009.06146.x
复制
发表时间：
2009-07
影响因子：
4.7
通讯作者：
Messing A
中科院分区：
文献类型：
--
作者：
Cho W;Hagemann TL;Johnson DA;Johnson JA;Messing A
胶质原纤维酸性蛋白（Glial fibrillary acid protein， GFAP）是星形胶质细胞主要的中间丝蛋白，其表达在发育过程和损伤后发生显著变化。为了便于研究GFAP的表达调控，我们培育了双转基因小鼠，分别在2.2 kb的人GFAP启动子调控下表达萤火虫荧光素酶和0.5 kb的人GAPDH启动子调控下表达Renilla荧光素酶，实现GFAP信号的归一化。与其他组织相比，gapdh - fluc在大脑中表达高，且仅限于星形胶质细胞，而GAPDH-RLuc表达更广泛。与单独使用GFAP信号相比，将GFAP信号归一化为GAPDH信号减少了个体间的变异性。GFAP/GAPDH比值正确反映了由于rd突变导致的FVB/N小鼠视网膜变性后GFAP的上调。kainic-acid诱导癫痫发作后，GFAP/GAPDH比值的变化先于GFAP总蛋白的变化。在表达R236H亚历山大病突变体的敲入小鼠中，GFAP启动子活性只是短暂升高，可能不能完全解释GFAP蛋白的积累。
Glial fibrillary acidic protein (GFAP) is the major intermediate filament protein of astrocytes, and its expression changes dramatically during development and following injury. To facilitate study of the regulation of GFAP expression, we have generated dual transgenic mice expressing both firefly luciferase under the control of a 2.2 kb human GFAP promoter and Renilla luciferase under the control of a 0.5 kb human GAPDH promoter for normalization of the GFAP signal. The GFAP-fLuc was highly expressed in brain compared to other tissues, and was limited to astrocytes, whereas the GAPDH-RLuc was more widely expressed. Normalization of the GFAP signal to the GAPDH signal reduced the inter-individual variability compared to using the GFAP signal alone. The GFAP/GAPDH ratio correctly reflected the up-regulation of GFAP that occurs following retinal degeneration in FVB/N mice due to the rd mutation. Following kainic-acid induced seizures, changes in the GFAP/GAPDH ratio precede those in total GFAP protein. In knock-in mice expressing the R236H Alexander disease mutant, GFAP promoter activity is only transiently elevated and may not entirely account for the accumulation of GFAP protein that takes place.
登录
查看更多内容
影响因子：
2.7
作者：
Kisseberth, WC;Brettingen, NT;Sandgren, EP
通讯作者：
Sandgren, EP
影响因子：
3.4
作者：
Calvo, Ana C.;Moreno-Igoa, Maria;Osta, Rosario
通讯作者：
Osta, Rosario
影响因子：
82.9
作者：
Contag, PR;Olomu, IN;Contag, CH
通讯作者：
Contag, CH
DOI：
10.1073/pnas.85.14.5092
发表时间：
1988-07-01
影响因子：
11.1
作者：
ALEXANDER, MC;LOMANTO, M;RAMAIKA, C
通讯作者：
RAMAIKA, C
DOI：
10.1016/0013-4694(72)90177-0
发表时间：
1972-01-01
期刊：
ELECTROENCEPHALOGRAPHY AND CLINICAL NEUROPHYSIOLOGY
影响因子：
--
作者：
RACINE, RJ
通讯作者：
RACINE, RJ