Silencing neurotransmission with membrane-tethered toxins
Silencing neurotransmission with membrane-tethered toxins
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DOI：
10.1038/nmeth.1425
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发表时间：
2010-03-01
期刊：
影响因子：
48
通讯作者：
Ibanez-Tallon, Ines
中科院分区：
文献类型：
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作者：
Auer, Sebastian;Stuerzebecher, Annika S.;Ibanez-Tallon, Ines
在突触终末，高电压激活的Ca（v）2.1和Ca（v）2.2钙通道在突触前动作电位与神经递质释放的偶联中具有重要的联合作用。在这里，我们表明，膜拴系毒素允许细胞自主封锁每个通道单独或同时在小鼠体内神经元。我们报告了优化的组成型，诱导型和Cre重组酶依赖性慢病毒载体编码荧光重组毒素，我们还验证了毒素为基础的策略在转基因小鼠模型。通过慢病毒载体递送的毒素选择性地抑制多巴胺能黑质纹状体通路，并且在伤害性外周神经元中靶向表达的转基因小鼠显示出对慢性疼痛的持久抑制。优化的拴系毒素是用于体内离子通道介导的活性的细胞特异性和时间操纵的工具，包括神经递质释放的阻断。
At synaptic terminals, high voltage-activated Ca(v)2.1 and Ca(v)2.2 calcium channels have an essential and joint role in coupling the presynaptic action potential to neurotransmitter release. Here we show that membrane-tethered toxins allowed cell-autonomous blockade of each channel individually or simultaneously in mouse neurons in vivo. We report optimized constitutive, inducible and Cre recombinase-dependent lentiviral vectors encoding fluorescent recombinant toxins, and we also validated the toxin-based strategy in a transgenic mouse model. Toxins delivered by lentiviral vectors selectively inhibited the dopaminergic nigrostriatal pathway, and transgenic mice with targeted expression in nociceptive peripheral neurons displayed long-lasting suppression of chronic pain. Optimized tethered toxins are tools for cell-specific and temporal manipulation of ion channel-mediated activities in vivo, including blockade of neurotransmitter release.