RRC ID 77379
Author Wang L, Wu C, Peng W, Zhou Z, Zeng J, Li X, Yang Y, Yu S, Zou Y, Huang M, Liu C, Chen Y, Li Y, Ti P, Liu W, Gao Y, Zheng W, Zhong H, Gao S, Lu Z, Ren PG, Ng HL, He J, Chen S, Xu M, Li Y, Chu J.
Title A high-performance genetically encoded fluorescent indicator for in vivo cAMP imaging.
Journal Nat Commun
Abstract cAMP is a key second messenger that regulates diverse cellular functions including neural plasticity. However, the spatiotemporal dynamics of intracellular cAMP in intact organisms are largely unknown due to low sensitivity and/or brightness of current genetically encoded fluorescent cAMP indicators. Here, we report the development of the new circularly permuted GFP (cpGFP)-based cAMP indicator G-Flamp1, which exhibits a large fluorescence increase (a maximum ΔF/F0 of 1100% in HEK293T cells), decent brightness, appropriate affinity (a Kd of 2.17 μM) and fast response kinetics (an association and dissociation half-time of 0.20 and 0.087 s, respectively). Furthermore, the crystal structure of the cAMP-bound G-Flamp1 reveals one linker connecting the cAMP-binding domain to cpGFP adopts a distorted β-strand conformation that may serve as a fluorescence modulation switch. We demonstrate that G-Flamp1 enables sensitive monitoring of endogenous cAMP signals in brain regions that are implicated in learning and motor control in living organisms such as fruit flies and mice.
Volume 13(1)
Pages 5363
Published 2022-9-12
DOI 10.1038/s41467-022-32994-7
PII 10.1038/s41467-022-32994-7
PMID 36097007
PMC PMC9468011
MeSH Animals Coloring Agents Diagnostic Imaging* Green Fluorescent Proteins / genetics HEK293 Cells Humans Mice Second Messenger Systems*
IF 12.121
Drosophila DGRC119070 DGRC119071