Visualizing estrogen receptor-α-expressing neurons using a new ERα-ZsGreen reporter mouse line.
Visualizing estrogen receptor-α-expressing neurons using a new ERα-ZsGreen reporter mouse line.
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DOI：
10.1016/j.metabol.2015.12.011
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发表时间：
2016-04
期刊：
影响因子：
--
通讯作者：
Xu Y
中科院分区：
文献类型：
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作者：
Saito K;He Y;Yan X;Yang Y;Wang C;Xu P;Hinton AO Jr;Shu G;Yu L;Tong Q;Xu Y
雌激素的多种生物学功能，包括能量代谢的调节，是由脑中表达雌激素受体-α（ERα）的神经元介导的。然而，由于缺乏可视化ERα表达神经元的策略，这些ERα表达神经元中复杂的细胞内过程难以解开，特别是在未固定的脑组织中。我们构建了一个新的ERα-ZS绿色报告基因小鼠系，其中绿色荧光报告蛋白ZS绿色的表达由241 kb的ERα基因启动子驱动。我们验证了ZsGreen与内源性ERα在脑中高度共定位。天然ZsGreen信号在未固定的脑组织中可视化，并用于辅助单细胞收集和电生理记录。最后，我们证明了这种ERα-ZsGreen小鼠等位基因可以与其他遗传报告等位基因组合使用，以允许在高度选择性的神经群体中进行实验。
A variety of biological functions of estrogens, including regulation of energy metabolism, are mediated by neurons expressing estrogen receptor-α (ERα) in the brain. However, complex intracellular processes in these ERα-expressing neurons are difficult to unravel, due to the lack of strategy to visualize ERα-expressing neurons, especially in unfixed brain tissues. Here we generated a novel ERα-ZsGreen reporter mouse line in which expression of a green fluorescent reporter protein, ZsGreen, is driven by a 241 kb ERα gene promoter. We validated that ZsGreen is highly colocalized with endogenous ERα in the brain. Native ZsGreen signals were visualized in unfixed brain tissue, and were used to assist single cell collection and electrophysiological recordings. Finally, we demonstrated that this ERα-ZsGreen mouse allele can be used in combination with other genetic reporter alleles to allow experiments in highly selective neural populations.