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多巴胺传感器扩展调色板助力活体多重成像
作者:小柯机器人 发布时间:2020/9/9 18:04:25

近日,美国加州大学教授Lin Tian,Joshua D. Berke,Mark von Zastrow 及其小组开发出红移的多巴胺体内成像传感器。 这一研究成果于2020年9月7日发表在《自然—方法学》上。

该课题组基于mApple设计了多巴胺传感器的红移变体,称为RdLight1。RdLight1可以与基于GFP的传感器结合使用,干扰最小,并且具有高光稳定性,可以实现长时间连续成像。

该课题组人员在以下体系中演示了RdLight1的效用:在体外培养的细胞中进行受体特异的药理学分析;在自由行为的老鼠上同时评估细胞类型特异的神经活动和多巴胺的释放以及同时次秒级监测多种神经递质。双色光度法解释了由奖赏预测信号(reward-predictive cues)诱发的在伏隔核的多巴胺释放是伴随着谷氨酸释放的快速抑制。通过支持活体的多重成像多巴胺及其他神经回路组成,RdLight1打开了多方面了解多巴胺生物学的途径。

研究人员表示,基于绿色荧光蛋白(GFP)的遗传编码的多巴胺传感器可对行为动物中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的变体无法与常用的光学传感器和执行器直接结合使用。

附:英文原文

Title: An expanded palette of dopamine sensors for multiplex imaging in vivo

Author: Tommaso Patriarchi, Ali Mohebi, Junqing Sun, Aaron Marley, Ruqiang Liang, Chunyang Dong, Kyle Puhger, Grace Or Mizuno, Carolyn M. Davis, Brian Wiltgen, Mark von Zastrow, Joshua D. Berke, Lin Tian

Issue&Volume: 2020-09-07

Abstract: 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. 

DOI: 10.1038/s41592-020-0936-3

Source: https://www.nature.com/articles/s41592-020-0936-3

期刊信息

Nature Methods:《自然—方法学》,创刊于2004年。隶属于施普林格·自然出版集团,最新IF:28.467
官方网址:https://www.nature.com/nmeth/
投稿链接:https://mts-nmeth.nature.com/cgi-bin/main.plex