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An expanded palette of dopamine sensors for multiplex imaging in vivo.
Nature Methods ( IF 36.1 ) Pub Date : 2020-09-07 , DOI: 10.1038/s41592-020-0936-3
Tommaso Patriarchi 1, 2, 3 , Ali Mohebi 4 , Junqing Sun 1, 5, 6 , Aaron Marley 7 , Ruqiang Liang 1 , Chunyang Dong 1 , Kyle Puhger 5 , Grace Or Mizuno 1 , Carolyn M Davis 1 , Brian Wiltgen 5 , Mark von Zastrow 7, 8 , Joshua D Berke 4, 8 , Lin Tian 1
Affiliation  

Genetically encoded dopamine sensors based on green fluorescent protein (GFP) enable high-resolution imaging of dopamine dynamics in behaving animals. However, these GFP-based variants cannot be readily combined with commonly used optical sensors and actuators, due to spectral overlap. We therefore engineered red-shifted variants of dopamine sensors called RdLight1, based on mApple. RdLight1 can be combined with GFP-based sensors with minimal interference and shows high photostability, permitting prolonged continuous imaging. We demonstrate the utility of RdLight1 for receptor-specific pharmacological analysis in cell culture, simultaneous assessment of dopamine release and cell-type-specific neuronal activity and simultaneous subsecond monitoring of multiple neurotransmitters in freely behaving rats. Dual-color photometry revealed that dopamine release in the nucleus accumbens evoked by reward-predictive cues is accompanied by a rapid suppression of glutamate release. By enabling multiplexed imaging of dopamine with other circuit components in vivo, RdLight1 opens avenues for understanding many aspects of dopamine biology.



中文翻译:

用于体内多重成像的多巴胺传感器的扩展调色板。

基于绿色荧光蛋白 (GFP) 的基因编码多巴胺传感器能够对行为动物的多巴胺动力学进行高分辨率成像。然而,由于光谱重叠,这些基于 GFP 的变体不能很​​容易地与常用的光学传感器和执行器结合使用。因此,我们基于 mApple 设计了称为 RdLight1 的多巴胺传感器的红移变体。RdLight1 可以与基于 GFP 的传感器结合使用,干扰最小,并且显示出高光稳定性,允许长时间连续成像。我们证明了 RdLight1 在细胞培养中的受体特异性药理学分析、多巴胺释放和细胞类型特异性神经元活动的同时评估以及自由行为大鼠中多种神经递质的同时亚秒级监测的效用。双色光度法显示,由奖励预测线索引起的伏隔核中的多巴胺释放伴随着谷氨酸释放的快速抑制。通过在体内实现多巴胺与其他电路组件的多重成像,RdLight1 为理解多巴胺生物学的许多方面开辟了道路。

更新日期:2020-09-08
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