Reference - Detail
| 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 |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | X(Twitter) |
| Total number of mentions | 16 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| Drosophila | DGRC#119070 DGRC#119071 |