Probing PAC1 receptor activation across species with an engineered sensor.

Autor: Cola RB; Institute of Pharmacology and Toxicology, University of Zürich, Zurich, Switzerland., Niethammer SN; Medical University of Vienna, Center for Brain Research, Department for Neuronal Cell Biology, Vienna, Austria., Rajamannar P; Department of Molecular Neuroscience & Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, Israel., Gresch A; Institute of Pharmacology and Toxicology, University of Zürich, Zurich, Switzerland., Bhat MA; Institute of Pharmacology and Toxicology, University of Zürich, Zurich, Switzerland., Assoumou K; Department of Cell Physiology and Metabolism, University of Geneva, Geneva, Switzerland., Williams ET; Department of Chemistry, University of Zürich, Zürich, Switzerland., Hauck P; Department of Chemistry, University of Zürich, Zürich, Switzerland., Hartrampf N; Department of Chemistry, University of Zürich, Zürich, Switzerland., Benke D; Institute of Pharmacology and Toxicology, University of Zürich, Zurich, Switzerland.; Neuroscience Center Zurich, University and ETH Zürich, Zürich, Switzerland., Stoeber M; Department of Cell Physiology and Metabolism, University of Geneva, Geneva, Switzerland., Levkowitz G; Department of Molecular Neuroscience & Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, Israel., Melzer S; Medical University of Vienna, Center for Brain Research, Department for Neuronal Cell Biology, Vienna, Austria., Patriarchi T; Institute of Pharmacology and Toxicology, University of Zürich, Zurich, Switzerland.; Neuroscience Center Zurich, University and ETH Zürich, Zürich, Switzerland.
Jazyk: angličtina
Zdroj: ELife [Elife] 2024 Aug 15; Vol. 13. Date of Electronic Publication: 2024 Aug 15.
DOI: 10.7554/eLife.96496
Abstrakt: Class-B1 G-protein-coupled receptors (GPCRs) are an important family of clinically relevant drug targets that remain difficult to investigate via high-throughput screening and in animal models. Here, we engineered PAClight1 P78A , a novel genetically encoded sensor based on a class-B1 GPCR (the human PAC1 receptor, hmPAC1R) endowed with high dynamic range (Δ F / F 0 = 1100%), excellent ligand selectivity, and rapid activation kinetics ( τ ON = 1.15 s). To showcase the utility of this tool for in vitro applications, we thoroughly characterized and compared its expression, brightness and performance between PAClight1 P78A -transfected and stably expressing cells. Demonstrating its use in animal models, we show robust expression and fluorescence responses upon exogenous ligand application ex vivo and in vivo in mice, as well as in living zebrafish larvae. Thus, the new GPCR-based sensor can be used for a wide range of applications across the life sciences empowering both basic research and drug development efforts.
Competing Interests: RC, SN, PR, AG, MB, KA, EW, PH, NH, DB, MS, GL, SM No competing interests declared, TP TP is a co-inventor on a patent application related to the technology described in this article
(© 2024, Cola et al.)
Databáze: MEDLINE
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