VAChT-Cre.Fast and VAChT-Cre.Slow: Postnatal expression of Cre recombinase in somatomotor neurons with different onset
VAChT-Cre.Fast and VAChT-Cre.Slow: Postnatal expression of Cre recombinase in somatomotor neurons with different onset
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DOI：
10.1002/gene.10224
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发表时间：
2003-09-01
期刊：
影响因子：
1.5
通讯作者：
Takahashi, R
中科院分区：
文献类型：
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作者：
Misawa, H;Nakata, K;Takahashi, R
胆碱能基因座（CGL）由编码胆碱乙酰转移酶（ChAT）和囊泡乙酰胆碱转运体（VAChT）的基因组成。为了建立胆碱能特异性表达Cre的小鼠，我们构建了一个含有11.3 kb人CGL的转基因表达载体（VAChT-Cre），其中Cre-IRES-EGFP单元插入到VAChT开放阅读框架中。Cre（其表达由VAChT启动子驱动）的活性通过杂交报告小鼠（CAG-CAT-Z）来检查，其中LacZ的表达在Cre介导的重组后被激活。VAChT-Cre构建体的转基因株系在体动核和内侧缰核的胆碱能神经元亚群中显示出有限的Cre表达，但在内脏和其他中枢和外周胆碱能神经元中不存在。Cre表达在出生后第7天首次观察到，随后在40 - 60%的躯体运动神经元中检测到。基于Cre表达的开始，我们产生了两个小鼠品系（两个等位基因; VAChT-Cre. Fast和VAChT-Cre. Slow），其中Cre表达分别快速和缓慢地达到最大水平。使用VAChT-Cre小鼠应该允许我们将Cre传递到出生后运动神经元的子集，从而绕过致死性并促进成年运动神经元中基因功能的分析。(C)2003 Wiley-Liss，Inc.
The cholinergic gene locus (CGL) consists of the genes encoding the choline acetyltransferase (ChAT) and the vesicular acetylcholine transporter (VAChT). To establish a cholinergic-specific Cre-expressing mouse, we constructed a transgene expression vector (VAChT-Cre) with 11.3 kb human CGL in which a Cre-IRES-EGFP unit was inserted in the VAChT open reading frame. The activity of Cre, whose expression was driven by the VAChT promoter, was examined by crossing a reporter mouse (CAG-CAT-Z) in which expression of LacZ is activated upon Cre-mediated recombination. Transgenic lines with the VAChT-Cre construct displayed the restricted Cre expression in a subset of cholinergic neurons in the somatomotor nuclei and medial habenular nucleus, but absent in visceromotor and other central and peripheral cholinergic neurons. Cre expression was first observed at postnatal day 7 and later detected in similar to40-60% of somatomotor neurons. Based on the onset of Cre expression, we generated two mouse lines (two alleles; VAChT-Cre.Fast and VAChT-Cre.Slow) in which Cre expression reaches maximal levels fast and slow, respectively. The use of VAChT-Cre mice should allow us to deliver Cre to a subset of postnatal motor neurons, thereby bypassing lethality and facilitating analysis of gene function in adult motor neurons. (C) 2003 Wiley-Liss, Inc.