INDICATOR EXPRESSION DIRECTED BY REGULATORY SEQUENCES OF THE GLIAL FIBRILLARY ACIDIC PROTEIN (GFAP) GENE - IN-VIVO COMPARISON OF DISTINCT GFAP-LACZ TRANSGENES
INDICATOR EXPRESSION DIRECTED BY REGULATORY SEQUENCES OF THE GLIAL FIBRILLARY ACIDIC PROTEIN (GFAP) GENE - IN-VIVO COMPARISON OF DISTINCT GFAP-LACZ TRANSGENES
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DOI：
10.1002/glia.440130304
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发表时间：
1995-03-01
期刊：
影响因子：
6.2
通讯作者：
MUCKE, L
中科院分区：
文献类型：
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作者：
JOHNSON, WB;RUPPE, MD;MUCKE, L
星形胶质细胞的胶质细胞酸性蛋白（GFAP）基因表达的增加似乎构成了大脑对损伤的反应的重要组成部分，因为它在许多不同的物种中可见，并且在各种神经系统疾病中表现突出。先前，我们已经使用修饰的GFAP基因（C-339）来将β-半乳糖苷酶（β-gal）的表达靶向转基因小鼠中的星形胶质细胞（Mucke等人; New Biol 3：465-474 1991）。为了确定基因内GFAP序列对GFAP驱动的融合基因在体内表达的影响程度，用E。将大肠杆菌lacZ报告基因置于约2kb鼠GFAP 5'侧翼区（C-259）的下游或连接到整个鼠GFAP基因（C-445）的外显子1中。表达C-259与C-445的转基因小鼠在其大脑中显示出相似的β-gal活性水平和分布。从GFAP-lacZ融合基因中排除基因内GFAP序列并没有减少损伤诱导的星形胶质细胞β-gal表达上调或增加非星形胶质细胞脑细胞中的β-gal表达。这些结果表明，2kb的鼠GFAP 5'侧翼区足以将转基因表达主要限制于星形胶质细胞并介导体内损伤反应性。因此，该序列构成了反应性星形胶质细胞增生介质的关键靶点。虽然急性穿透性脑损伤在C-259、C-445和C-339转基因小鼠的病变部位周围诱导了β-gal表达的局灶性增加，但用羊瘙痒病感染C-339转基因小鼠导致星形胶质细胞β-gal表达的广泛上调。因此，GFAP-lacZ转基因小鼠可用于监测体内星形胶质细胞活化的差异模式。这些和相关的模型应有助于评估的策略，旨在在体内操纵GFAP表达和星形胶质细胞活化。(C)1995 Wiley-Liss，Inc.
An increase in the expression of the glial fibrillary acidic protein (GFAP) gene by astrocytes appears to constitute a crucial component of the brain's response to injury because it is seen in many different species and features prominently in diverse neurological diseases. Previously, we have used a modified GFAP gene (C-339) to target the expression of beta-galactosidase (beta-gal) to astrocytes in transgenic mice (Mucke et al.; New Biol 3:465-474 1991). To determine to what extent the in vivo(1) expression of GFAP-driven fusion genes is influenced by intragenic GFAP sequences, the E. coli lacZ reporter gene was either placed downstream of approximately 2 kb of murine GFAP 5' flanking region (C-259) or ligated into exon 1 of the entire murine GFAP gene (C-445). Transgenic mice expressing C-259 versus C-445 showed similar levels and distributions of beta-gal activity in their brains. Exclusion of intragenic GFAP sequences from the GFAP-lacZ fusion gene did not diminish injury-induced upmodulation of astroglial beta-gal expression or increase beta-gal expression in non-astrocytic brain cells. These results demonstrate that 2 kb of murine GFAP 5' flanking region is sufficient to restrict transgene expression primarily to astrocytes and to mediate injury-responsiveness in vivo. This sequence therefore constitutes a critical target for mediators of reactive astrocytosis. While acute penetrating brain injuries induced focal increases in beta-gal expression around the lesion sites in C-259, C-445, and C-339 transgenic mice, infection of C-339 transgenic mice with scrapie led to a widespread upmodulation of astroglial beta-gal expression. Hence, GFAP-lacZ transgenic mice can be used to monitor differential patterns of astroglial activation in vivo. These and related models should facilitate the assessment of strategies aimed at the in vive manipulation of GFAP expression and astroglial activation. (C) 1995 Wiley-Liss, Inc.