Abstract
SUMMARYcAMP is a pivotal second messenger regulated by various upstream pathways including Ca2+and G protein-coupled receptors (GPCRs). To decipherin vivocAMP dynamics, we rationally designed cAMPinG1, an ultrasensitive 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 highly sensitive new red Ca2+indicator RCaMP3 allowed simultaneous measurement of population patterns in Ca2+and cAMP in hundreds of neurons. We identified Ca2+-induced cAMP responses that represented specific information, such as direction selectivity in vision and locomotion, as well as GPCR-induced cAMP responses. Overall, our multicolor suite revealed that information encoded in Ca2+and GPCRs signaling is integrated and stored as cAMP transients for longer periodsin vivo.HighlightsDeveloping an ultrasensitive cAMP indicator, cAMPinG1, for visualizing cAMP transients in somata and dendritic spinesin vivo.Developing a highly sensitive red Ca2+indicator, RCaMP3, for visualizing Ca2+transients in large neuronal population.Dual-color Ca2+and cAMP imaging for dissecting Ca2+-induced and GPCR-induced cAMP responses.Single-cell, single-timepoint cAMP imaging for GRCR biology and drug screening.
Publisher
Cold Spring Harbor Laboratory
Cited by
2 articles.
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