Light generation of intracellular Ca(2+) signals by a genetically encoded protein BACCS.
Light generation of intracellular Ca(2+) signals by a genetically encoded protein BACCS.
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DOI：
10.1038/ncomms9021
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发表时间：
2015-08-18
影响因子：
16.6
通讯作者：
Nakata T
中科院分区：
文献类型：
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作者：
Ishii T;Sato K;Kakumoto T;Miura S;Touhara K;Takeuchi S;Nakata T
Ca2+信号在许多细胞生理事件中以时空方式受到高度调节。在这里，我们报告了一个基因工程蓝光激活的Ca 2+通道开关（BACCS），作为光遗传学工具产生Ca 2+信号。BACCS响应于光打开Ca 2+选择性奥赖离子通道。BACCS变体dmBACCS 2与果蝇奥赖组合更迅速地升高Ca 2+浓度，使得在光暴露1秒内观察到哺乳动物细胞中的Ca 2+升高。使用BACCS，我们成功地控制细胞事件，包括NFAT介导的基因表达。在小鼠嗅觉系统中，BACCS介导光依赖性电生理反应。此外，我们产生BACCS突变体，其表现出细胞内Ca 2+的快速和缓慢恢复。因此，BACCS是用于产生具有大动态范围的时间上不同的细胞内Ca 2+信号的有用的光遗传学工具，并且将适用于体外和体内研究。目前用于细胞内钙信号的光遗传学控制的工具目前遭受慢响应时间或低动态范围。在这里，作者开发了蓝光激活的Ca 2+通道开关（BACCS），其以高时间分辨率和大动态范围调节Ca 2+敏感的奥赖通道的活性。
Ca2+ signals are highly regulated in a spatiotemporal manner in numerous cellular physiological events. Here we report a genetically engineered blue light-activated Ca2+ channel switch (BACCS), as an optogenetic tool for generating Ca2+ signals. BACCS opens Ca2+-selective ORAI ion channels in response to light. A BACCS variant, dmBACCS2, combined with Drosophila Orai, elevates the Ca2+ concentration more rapidly, such that Ca2+ elevation in mammalian cells is observed within 1 s on light exposure. Using BACCSs, we successfully control cellular events including NFAT-mediated gene expression. In the mouse olfactory system, BACCS mediates light-dependent electrophysiological responses. Furthermore, we generate BACCS mutants, which exhibit fast and slow recovery of intracellular Ca2+. Thus, BACCSs are a useful optogenetic tool for generating temporally various intracellular Ca2+ signals with a large dynamic range, and will be applicable to both in vitro and in vivo studies. Current tools for optogenetic control of intracellular calcium signals currently suffer from slow response time or low dynamic range. Here the authors develop blue light-activated Ca2+ channel switch (BACCS) that modulates the activity of Ca2+-sensitive Orai channels with high temporal resolution and large dynamic range.