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A fluorescent sensor for spatiotemporally resolved endocannabinoid dynamics in vitro and in vivo

Ao Dong, Kaikai He, Barna Dudok, Jordan S Farrell, Wuqiang Guan, Daniel J Liput, Henry L Puhl, Ruyi Cai, Jiali Duan, Eddy Albarran, Jun Ding, David M Lovinger, Bo Li, Ivan Soltesz, Yulong Li
doi: https://doi.org/10.1101/2020.10.08.329169
Ao Dong
1State Key Laboratory of Membrane Biology, Peking University School of Life Sciences, Beijing, China
2PKU-IDG/McGovern Institute for Brain Research, Beijing, China
3Peking-Tsinghua Center for Life Sciences, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing, China
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Kaikai He
1State Key Laboratory of Membrane Biology, Peking University School of Life Sciences, Beijing, China
2PKU-IDG/McGovern Institute for Brain Research, Beijing, China
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Barna Dudok
4Department of Neurosurgery, Stanford University, Palo Alto, CA, USA
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Jordan S Farrell
4Department of Neurosurgery, Stanford University, Palo Alto, CA, USA
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Wuqiang Guan
5Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, USA
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Daniel J Liput
6Laboratory for Integrative Neuroscience, National Institute on Alcohol Abuse and Alcoholism, Bethesda, MD, USA
7Laboratory of Molecular Physiology, National Institute on Alcohol Abuse and Alcoholism, Bethesda, MD, USA
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Henry L Puhl
7Laboratory of Molecular Physiology, National Institute on Alcohol Abuse and Alcoholism, Bethesda, MD, USA
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Ruyi Cai
1State Key Laboratory of Membrane Biology, Peking University School of Life Sciences, Beijing, China
2PKU-IDG/McGovern Institute for Brain Research, Beijing, China
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Jiali Duan
1State Key Laboratory of Membrane Biology, Peking University School of Life Sciences, Beijing, China
2PKU-IDG/McGovern Institute for Brain Research, Beijing, China
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Eddy Albarran
8Neuroscience PhD Program, Stanford University, Palo Alto, CA, USA
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Jun Ding
9Department of Neurosurgery, Department of Neurology, Stanford University School of Medicine, Palo Alto, CA, USA
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David M Lovinger
6Laboratory for Integrative Neuroscience, National Institute on Alcohol Abuse and Alcoholism, Bethesda, MD, USA
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Bo Li
5Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, USA
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Ivan Soltesz
4Department of Neurosurgery, Stanford University, Palo Alto, CA, USA
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Yulong Li
1State Key Laboratory of Membrane Biology, Peking University School of Life Sciences, Beijing, China
2PKU-IDG/McGovern Institute for Brain Research, Beijing, China
3Peking-Tsinghua Center for Life Sciences, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing, China
10Chinese Institute for Brain Research, Beijing, China
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  • For correspondence: yulongli@pku.edu.cn
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Abstract

Endocannabinoids (eCBs) are retrograde neuromodulators that play an important role in a wide range of physiological processes; however, the release and in vivo dynamics of eCBs remain largely unknown, due in part to a lack of suitable probes capable of detecting eCBs with sufficient spatiotemporal resolution. Here, we developed a new eCB sensor called GRABeCB2.0. This genetically encoded sensor consists of the human CB1 cannabinoid receptor fused to circular-permutated EGFP, providing cell membrane trafficking, second-resolution kinetics, high specificity for eCBs, and a robust fluorescence response at physiological eCB concentrations. Using the GRABeCB2.0 sensor, we monitored evoked changes in eCB dynamics in both cultured neurons and acute brain slices. Interestingly, in cultured neurons we also observed spontaneous compartmental eCB transients that spanned a distance of approximately 11 μm, suggesting constrained, localized eCB signaling. Moreover, by expressing GRABeCB2.0 in the mouse brain, we readily observed foot shock-elicited and running-triggered eCB transients in the basolateral amygdala and hippocampus, respectively. Lastly, we used GRABeCB2.0 in a mouse seizure model and observed a spreading wave of eCB release that followed a Ca2+ wave through the hippocampus. Thus, GRABeCB2.0 is a robust new probe for measuring the dynamics of eCB release under both physiological and pathological conditions.

Competing Interest Statement

Y. L. has filed patent applications, the value of which might be affected by this publication.

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Posted October 20, 2020.
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A fluorescent sensor for spatiotemporally resolved endocannabinoid dynamics in vitro and in vivo
Ao Dong, Kaikai He, Barna Dudok, Jordan S Farrell, Wuqiang Guan, Daniel J Liput, Henry L Puhl, Ruyi Cai, Jiali Duan, Eddy Albarran, Jun Ding, David M Lovinger, Bo Li, Ivan Soltesz, Yulong Li
bioRxiv 2020.10.08.329169; doi: https://doi.org/10.1101/2020.10.08.329169
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A fluorescent sensor for spatiotemporally resolved endocannabinoid dynamics in vitro and in vivo
Ao Dong, Kaikai He, Barna Dudok, Jordan S Farrell, Wuqiang Guan, Daniel J Liput, Henry L Puhl, Ruyi Cai, Jiali Duan, Eddy Albarran, Jun Ding, David M Lovinger, Bo Li, Ivan Soltesz, Yulong Li
bioRxiv 2020.10.08.329169; doi: https://doi.org/10.1101/2020.10.08.329169

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