Abstract
Muscarinic acetylcholine receptor subtype 3 (M3 receptor) is a G Protein-Coupled Receptor (GPCR) that mediates many important physiological functions. Currently, most M3 receptor drugs also have high affinity for other subtypes of muscarinic acetylcholine receptors (mAChRs) and produce the risk of side effects. Therefore, in order to find M3 receptor drugs with high specificity, high activity and low side effects, we established a cell model and method for efficient and sensitive screening of M3 receptor based on calcium-activated chloride channels (CaCCs), and this method is also suitable for the screening of other GPCR drugs. This screening model consists of Fischer rat thyroid follicular epithelial (FRT) cells that endogenously express M3 receptors, CaCCs, and the indicator YFP-H148Q/I152L. We verified that the model can sensitively detect changes in intracellular Ca2+ concentration using fluorescence quenching kinetics experiments, confirmed the screening function of the model by applying available M3 receptor drugs, and also evaluated the good performance of the model in high-throughput screening.
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Acknowledgements
This work was partly supported by the National Natural Science Foundation of China (81601234), the Science and Technology Project of Traditional Chinese Medicine of Jilin Province (2021092), the Starting fund for doctoral research of Jilin Medical University (JYBS2021013lk), the Project of Jilin Provincial Department of Education (JJKH20220462KJ, JJKH20220473SK), the Project of Science and Technology Department of Jilin Province (20220101338JC), the “Chenyu” Research and Innovation Project of Beihua University School of Medical Technology (2021006), the Project of Beihua University Graduate Innovation Program (2022021).
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Liu, X., Ju, X., Hong, Q. et al. Establishment of a CaCC-based Cell Model and Method for High-throughput Screening of M3 Receptor Drugs. Cell Biochem Biophys 81, 49–58 (2023). https://doi.org/10.1007/s12013-022-01119-8
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DOI: https://doi.org/10.1007/s12013-022-01119-8