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
DNA material pN1-cAMPinG1 (RDB19691) pAAV-eSyn-cAMPinG1-NE-WPRE (RDB19692) pAAV-eSyn-cAMPinG1-ST-WPRE (RDB19693) pAAV-eSyn-RCaMP3-WPRE (RDB19694)