{"doi":"10.1101/2025.04.30.650917","title":"Multi-parametric imaging of spatio-temporal cAMP signaling, transmembrane potential, and intracellular calcium in the Langendorff-perfused heart","abstract":"Summary The spatiotemporal dynamics of intracellular second messengers and signaling molecules, including cyclic adenosine monophosphate (cAMP), have been studied extensively in isolated cardiomyocytes using Förster resonance energy transfer (FRET)-based reporters. However, little is known about how molecular signaling events affect tissue-level function. Optical mapping of transmembrane potential (V m ) and intracellular calcium (Ca 2+ i ) is frequently performed in isolated intact hearts to reveal tissue-level electrophysiological function but cannot reveal molecular underpinnings. Here, we developed a novel multi-parametric optical imaging system that enables multi-parametric recording of four wavelengths to visualize cAMP and real-time electrophysiological responses (V m and Ca 2+ i ) by utilizing concurrent FRE T imaging and d ual o ptical m apping (FRE-DOM) in the intact heart from a cardiac-specific FRET-based cAMP reporter mouse. We showed that cAMP is strongly and heterogeneously activated throughout the heart (atria and ventricles) in response to β-AR stimulation, the time course of which matches heart rate, action potential, and Ca 2+ responses. This novel imaging system will provide insight into the relationship between cAMP signaling, electrophysiology, and arrhythmogenesis at high spatio-temporal resolution.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2025,"id":565478,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":0,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9538,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2025-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":783858,"name":"Jessica L. Caldwell","orcid":"0000-0002-8616-318X","position":1,"is_corresponding":false},{"id":1468859,"name":"Lilian Mott","orcid":"0000-0003-3144-8820","position":2,"is_corresponding":false},{"id":338677,"name":"Lianguo Wang","orcid":"0000-0002-4747-0419","position":3,"is_corresponding":false},{"id":338682,"name":"Crystal M. Ripplinger","orcid":"0000-0002-9908-4106","position":4,"is_corresponding":false},{"id":1055405,"name":"I-Ju Lee","orcid":"0000-0001-9822-6005","position":0,"is_corresponding":true}],"reference_count":37,"raw_metadata":null,"created_at":"2026-07-19T02:56:28.709936Z","pmid":"40654741","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}