Sensitivity optimization of a rhodopsin-based fluorescent voltage indicator
Sensitivity optimization of a rhodopsin-based fluorescent voltage indicator
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DOI：
10.1016/j.neuron.2023.03.009
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发表时间：
2023-05-17
期刊：
影响因子：
16.2
通讯作者：
Kolb, Ilya
中科院分区：
文献类型：
--
作者：
Abdelfattah, Ahmed S.;Zheng, Jihong;Kolb, Ilya
能够以毫秒级的分辨率对细胞跨膜电压进行光学成像的能力，可以提供对行为动物活的大脑功能的前所未有的洞察。在这里，我们提出了一种点突变，它提高了基于ACE2光学蛋白的电压指示器的灵敏度。我们使用突变来开发Voltron2，这是一种改进的化学电压指示器，对单个AP的敏感度比Voltron高65%，对亚阈值电位的敏感度比Voltron高3倍。Voltron2保留了其前身的亚毫秒动力学和光稳定性，尽管具有较低的基线荧光。在多个物种的体外和体内与其前身的多次比较中，我们发现Voltron2对AP和亚阈值波动更敏感。最后，我们利用Voltron2研究和评估了小鼠海马神经元间同步化的可能机制。总体而言，我们发现了一种可推广的突变，它显著提高了基于ACE2视紫红质的传感器的灵敏度，改善了它们的电压报告能力。
The ability to optically image cellular transmembrane voltages at millisecond-timescale resolutions can offer un-precedented insight into the function of living brains in behaving animals. Here, we present a point mutation that increases the sensitivity of Ace2 opsin-based voltage indicators. We use the mutation to develop Voltron2, an improved chemigeneic voltage indicator that has a 65% higher sensitivity to single APs and 3-fold higher sensi-tivity to subthreshold potentials than Voltron. Voltron2 retained the sub-millisecond kinetics and photostability of its predecessor, although with lower baseline fluorescence. In multiple in vitro and in vivo comparisons with its predecessor across multiple species, we found Voltron2 to be more sensitive to APs and subthreshold fluctu-ations. Finally, we used Voltron2 to study and evaluate the possible mechanisms of interneuron synchronization in the mouse hippocampus. Overall, we have discovered a generalizable mutation that significantly increases the sensitivity of Ace2 rhodopsin-based sensors, improving their voltage reporting capability.