Expression Patterns of Inducible Cre Recombinase Driven by Differential Astrocyte-Specific Promoters in Transgenic Mouse Lines
Expression Patterns of Inducible Cre Recombinase Driven by Differential Astrocyte-Specific Promoters in Transgenic Mouse Lines
复制标题
转基因小鼠系中差异星形胶质细胞特异性启动子驱动的诱导型 Cre 重组酶的表达模式。
DOI：
10.1007/s12264-019-00451-z
复制
发表时间：
2020-05-01
影响因子：
5.6
通讯作者：
Gao, Tian-Ming
中科院分区：
文献类型：
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作者：
Hu, Neng-Yuan;Chen, Ya-Ting;Gao, Tian-Ming
星形胶质细胞是中枢神经系统（CNS）中最丰富的细胞类型。它们为神经元提供营养支持，调节突触传递和可塑性，并导致神经元功能障碍。已经产生了许多转基因小鼠品系，以获得星形胶质细胞特异性表达的诱导型Cre重组酶的功能研究，然而，诱导型Cre重组酶在这些品系中的表达模式尚未得到系统的表征。我们建立了一个新的星形胶质细胞特异性Aldh 1 l1-CreER（T2）基因敲入小鼠品系，并比较了Cre重组酶在该品系和五个广泛使用的转基因品系之间的表达模式（来自杰克逊实验室和突变小鼠资源和研究中心的hGfap-CreER（T2）、Glast-CreER（T2）、Cx 30-CreER（T2）和Fgfr 3-iCreER（T2）），其在Cre介导的重组后表达tdTomato荧光。在成年Aldh 1 l1-CreER（T2）：Ai 14转基因小鼠中，在整个CNS中检测到tdTomato，并描述了五种新的形态学定义类型的星形胶质细胞。在六个评估的线，Cre介导的重组的特异性是最高的驱动时，Aldh 1 l1和最低的驱动时，hGFAP;在后者的小鼠，tdTomato和NeuN之间的共染色观察到在海马和皮质。值得注意的是，在Fgfr 3-iCreER（T2）小鼠中注意到明显的渗漏，并且当Cre重组酶表达由Glast驱动时，tdTomato的表达水平在丘脑中低，并且当由Cx 30驱动时，在中央杏仁核的被膜部分中低。此外，tdTomato在四个品系的外周器官中明确表达。我们的研究结果强调，星形胶质细胞特异性CreER（T2）转基因株系的功能研究中使用，应仔细选择。
Astrocytes are the most abundant cell type in the central nervous system (CNS). They provide trophic support for neurons, modulate synaptic transmission and plasticity, and contribute to neuronal dysfunction. Many transgenic mouse lines have been generated to obtain astrocyte-specific expression of inducible Cre recombinase for functional studies; however, the expression patterns of inducible Cre recombinase in these lines have not been systematically characterized. We generated a new astrocyte-specific Aldh1l1-CreER(T2) knock-in mouse line and compared the expression pattern of Cre recombinase between this and five widely-used transgenic lines (hGfap-CreER(T2) from The Jackson Laboratory and The Mutant Mouse Resource and Research Center, Glast-CreER(T2), Cx30-CreER(T2), and Fgfr3-iCreER(T2)) by crossing with Ai14 mice, which express tdTomato fluorescence following Cre-mediated recombination. In adult Aldh1l1-CreER(T2):Ai14 transgenic mice, tdTomato was detected throughout the CNS, and five novel morphologically-defined types of astrocyte were described. Among the six evaluated lines, the specificity of Cre-mediated recombination was highest when driven by Aldh1l1 and lowest when driven by hGfap; in the latter mice, co-staining between tdTomato and NeuN was observed in the hippocampus and cortex. Notably, evident leakage was noted in Fgfr3-iCreER(T2) mice, and the expression level of tdTomato was low in the thalamus when Cre recombinase expression was driven by Glast and in the capsular part of the central amygdaloid nucleus when driven by Cx30. Furthermore, tdTomato was clearly expressed in peripheral organs in four of the lines. Our results emphasize that the astrocyte-specific CreER(T2) transgenic lines used in functional studies should be carefully selected.