An expanded palette of dopamine sensors for multiplex imaging in vivo.
An expanded palette of dopamine sensors for multiplex imaging in vivo.
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DOI：
10.1038/s41592-020-0936-3
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发表时间：
2020-11
期刊：
影响因子：
48
通讯作者：
Tian L
中科院分区：
文献类型：
--
作者：
Patriarchi T;Mohebi A;Sun J;Marley A;Liang R;Dong C;Puhger K;Mizuno GO;Davis CM;Wiltgen B;von Zastrow M;Berke JD;Tian L
基于绿色荧光蛋白(GFP)的基因编码的多巴胺传感器能够对行为动物的多巴胺动态进行高分辨率成像。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相结合。因此，我们设计了红移的多巴胺传感器变种，称为RdLight1，基于MApple。RdLight1可以与基于GFP的传感器结合在一起，干扰最小，并显示出高的光稳定性，允许长时间连续成像。我们展示了RdLight1在细胞培养中的受体特异性药理学分析，同时评估多巴胺释放和细胞类型特异性神经元活动，以及在自由行为的大鼠中同步亚秒监测多种神经递质的效用。双色光度法显示，伏隔核多巴胺的释放在奖赏预测线索的刺激下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路成像，RdLight1为了解多巴胺生物学的许多方面开辟了道路。红色和黄色版本的遗传编码的多巴胺传感器dLight1已经被开发出来，允许多巴胺与神经递质或细胞类型的特定钙结合绿色传感器或致动器的多路成像，如在体外和行为啮齿动物中所展示的那样。
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. Red and yellow versions of the genetically encoded dopamine sensor dLight1 have been developed and allow multiplexed imaging of dopamine with neurotransmitter or cell-type specific calcium combined with green sensors or actuators as demonstrated ex vivo and in behaving rodents.
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影响因子：
3.7
作者：
Burgos-Robles A;Bravo-Rivera H;Quirk GJ
通讯作者：
Quirk GJ
影响因子：
21.1
作者：
Chen, Yingxiao;Song, Xianqiang;Ji, Guangju
通讯作者：
Ji, Guangju
影响因子：
64.8
作者：
Eichel K;Jullié D;Barsi-Rhyne B;Latorraca NR;Masureel M;Sibarita JB;Dror RO;von Zastrow M
通讯作者：
von Zastrow M
影响因子：
16.2
作者：
Hnasko, Thomas S.;Chuhma, Nao;Zhang, Hui;Goh, Germaine Y.;Sulzer, David;Palmiter, Richard D.;Rayport, Stephen;Edwards, Robert H.
通讯作者：
Edwards, Robert H.
影响因子：
16.2
作者：
Feng, Jiesi;Zhang, Changmei;Li, Yulong
通讯作者：
Li, Yulong