Targeted gene expression in dopamine and serotonin neurons of the mouse brain
Targeted gene expression in dopamine and serotonin neurons of the mouse brain
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DOI：
10.1016/j.jneumeth.2004.09.020
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发表时间：
2005-04-15
影响因子：
3
通讯作者：
Hen, R
中科院分区：
文献类型：
--
作者：
Zhuang, XX;Masson, J;Hen, R
我们使用了敲入策略，以产生两个线的小鼠表达Cre重组酶的转录控制下的多巴胺转运蛋白启动子（DAT-cre小鼠）或5-羟色胺转运蛋白启动子（SERT-cre小鼠）。在DAT-cre小鼠，免疫细胞化学染色的多巴胺合成酶酪氨酸羟化酶和Cre重组酶的成年大脑显示，几乎所有的多巴胺能神经元在腹侧中脑表达Cre。将DAT-cre小鼠与ROSA 26-stop-lacZ或ROSA 26-stop-YFP报告小鼠杂交，揭示了酪氨酸羟化酶和β-半乳糖苷酶或YFP染色之间近乎完美的相关性。YFP标记的荧光多巴胺能神经元可以很容易地在活体切片中识别。将SERT-cre小鼠与ROSA 26-stop-lacZ或ROSA 26-stop-YFP报告小鼠杂交，类似地揭示了马槟榔素合成酶色氨酸羟化酶和β-半乳糖苷酶或YFP的染色之间近乎完美的相关性。额外的Cre表达在丘脑和皮质中观察到，反映了已知的模式，短暂的SERT表达在出生后早期的发展。这些研究结果表明，使用神经递质转运蛋白启动子驱动选择性Cre表达，从而控制特定神经递质系统的突变的一般策略。与荧光基因报告交叉，这种策略通过神经递质状态标记神经元，为电生理学和成像提供了新的工具。(c)2004 Elsevier B. V.保留所有权利。
We used a knock-in strategy to generate two lines of mice expressing Cre recombinase under the transcriptional control of the dopamine transporter promoter (DAT-cre mice) or the serotonin transporter promoter (SERT-cre mice). In DAT-cre mice, immunocytochemical staining of adult brains for the dopamine-synthetic enzyme tyrosine hydroxylase and for Cre recombinase revealed that virtually all dopaminergic neurons in the ventral midbrain expressed Cre. Crossing DAT-cre mice with ROSA26-stop-lacZ or ROSA26-stop-YFP reporter mice revealed a near perfect correlation between staining for tyrosine hydroxylase and beta-galactosidase or YFP. YFP-labeled fluorescent dopaminergic neurons could be readily identified in live slices. Crossing SERT-cre mice with the ROSA26-stop-lacZ or ROSA26-stop-YFP reporter mice similarly revealed a near perfect correlation between staining for serotonin-synthetic enzyme tryptophan hydroxylase and beta-galactosidase or YFP. Additional Cre expression in the thalamus and cortex was observed, reflecting the known pattern of transient SERT expression during early postnatal development. These findings suggest a general strategy of using neurotransmitter transporter promoters to drive selective Cre expression and thus control mutations in specific neurotransmitter systems. Crossed with fluorescent-gene reporters, this strategy tags neurons by neurotransmitter status, providing new tools for electrophysiology and imaging. (c) 2004 Elsevier B.V. All rights reserved.