A fluorescent sensor for spatiotemporally resolved imaging of endocannabinoid dynamics in vivo

Dong, Ao, He, Kaikai, Dudok, Barna, Farrell, Jordan S, Guan, Wuqiang, Liput, Daniel J, Puhl, Henry L, Cai, Ruyi, Wang, Huan, Duan, Jiali, Albarran, Eddy, Ding, Jun, Lovinger, David M, Li, Bo, Soltesz, Ivan, Li, Yulong (November 2021) A fluorescent sensor for spatiotemporally resolved imaging of endocannabinoid dynamics in vivo. Nature Biotechnology. ISSN 1087-0156

URL: https://www.ncbi.nlm.nih.gov/pubmed/34764491
DOI: 10.1038/s41587-021-01074-4


Endocannabinoids (eCBs) are retrograde neuromodulators with important functions in a wide range of physiological processes, but their in vivo dynamics remain largely uncharacterized. Here we developed a genetically encoded eCB sensor called GRABeCB2.0. GRABeCB2.0 consists of a circular-permutated EGFP and the human CB1 cannabinoid receptor, providing cell membrane trafficking, second-resolution kinetics with high specificity for eCBs, and shows a robust fluorescence response at physiological eCB concentrations. Using GRABeCB2.0, we monitored evoked and spontaneous changes in eCB dynamics in cultured neurons and acute brain slices. We observed spontaneous compartmentalized eCB transients in cultured neurons and eCB transients from single axonal boutons in acute brain slices, suggesting constrained, localized eCB signaling. When GRABeCB2.0 was expressed in the mouse brain, we observed foot shock-elicited and running-triggered eCB signaling in the basolateral amygdala and hippocampus, respectively. In a mouse model of epilepsy, we observed a spreading wave of eCB release that followed a Ca2+ wave through the hippocampus. GRABeCB2.0 is a robust probe for eCB release in vivo.

Item Type: Paper
Subjects: organism description > animal behavior
Investigative techniques and equipment > imaging
CSHL Authors:
Communities: CSHL labs > Li lab
SWORD Depositor: CSHL Elements
Depositing User: CSHL Elements
Date: 11 November 2021
Date Deposited: 17 Nov 2021 16:09
Last Modified: 16 May 2022 13:34
PMCID: PMC9091059
URI: https://repository.cshl.edu/id/eprint/40422

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