| RRC ID |
80045
|
| Author |
Yokoyama T, Manita S, Uwamori H, Tajiri M, Imayoshi I, Yagishita S, Murayama M, Kitamura K, Sakamoto M.
|
| Title |
A multicolor suite for deciphering population coding of calcium and cAMP in vivo.
|
| Journal |
Nat Methods
|
| Abstract |
cAMP is a universal second messenger regulated by various upstream pathways including Ca2+ and G-protein-coupled receptors (GPCRs). To decipher in vivo cAMP dynamics, we rationally designed cAMPinG1, a sensitive genetically encoded green cAMP indicator that outperformed its predecessors in both dynamic range and cAMP affinity. Two-photon cAMPinG1 imaging detected cAMP transients in the somata and dendritic spines of neurons in the mouse visual cortex on the order of tens of seconds. In addition, multicolor imaging with a sensitive red Ca2+ indicator RCaMP3 allowed simultaneous measurement of population patterns in Ca2+ and cAMP in hundreds of neurons. We found Ca2+-related cAMP responses that represented specific information, such as direction selectivity in vision and locomotion, as well as GPCR-related cAMP responses. Overall, our multicolor suite will facilitate analysis of the interaction between the Ca2+, GPCR and cAMP signaling at single-cell resolution both in vitro and in vivo.
|
| Volume |
21(5)
|
| Pages |
897-907
|
| Published |
2024-5-1
|
| DOI |
10.1038/s41592-024-02222-9
|
| PII |
10.1038/s41592-024-02222-9
|
| PMID |
38514778
|
| PMC |
PMC11093745
|
| MeSH |
Animals
Calcium* / metabolism
Calcium Signaling
Cyclic AMP* / metabolism
HEK293 Cells
Humans
Mice
Mice, Inbred C57BL
Neurons* / metabolism
Receptors, G-Protein-Coupled / metabolism
Visual Cortex* / cytology
Visual Cortex* / metabolism
Visual Cortex* / physiology
|
| Resource |
| DNA material |
pN1-cAMPinG1 (RDB19691)
pAAV-eSyn-cAMPinG1-NE-WPRE (RDB19692)
pAAV-eSyn-cAMPinG1-ST-WPRE (RDB19693)
pAAV-eSyn-RCaMP3-WPRE (RDB19694) |
