Ciaramellano Francesca, Leuti Alessandro, Kurtz Alexandrine D E, Sarott Roman, Westphal Matthias, Pfaff Patrick, Grether Uwe, Carreira Erick M, Maccarrone Mauro, Oddi Sergio
European Center for Brain Research/Institute for Research and Health Care (IRCCS) Santa Lucia Foundation, Via del Fosso di Fiorano 64, 00143, Rome, Italy.
Department of Medicine, Campus Bio-Medico University of Rome, Rome, Italy.
Chembiochem. 2025 Apr 14;26(8):e202400921. doi: 10.1002/cbic.202400921. Epub 2025 Mar 12.
In this study, we employed a novel fluorescent probe, RO7304924 - which selectively targets cannabinoid 2 receptor (CBR) - to assess the lateral mobility of CBR within the plasma membrane of Chinese hamster ovary cells stably expressing a functional, untagged receptor variant. Utilizing confocal fluorescence recovery after photobleaching (FRAP), we quantified the diffusion coefficient and mobile fraction of CBR, thereby demonstrating the efficacy of RO7304924 as an innovative tool for elucidating the dynamics of this major endocannabinoid-binding G protein-coupled receptor. Our present findings highlight the potential of combining advanced ligand-based fluorescent probes with FRAP for future investigations into the biochemical details of CBR mobility in living cells, and its impact on receptor-dependent cellular processes.
在本研究中，我们使用了一种新型荧光探针RO7304924（其选择性靶向大麻素2受体（CBR）），以评估CBR在稳定表达功能性、无标签受体变体的中国仓鼠卵巢细胞膜内的侧向流动性。利用共聚焦光漂白后荧光恢复（FRAP）技术，我们对CBR的扩散系数和可移动部分进行了量化，从而证明了RO7304924作为一种创新工具，用于阐明这种主要内源性大麻素结合G蛋白偶联受体动力学的有效性。我们目前的研究结果突出了将先进的基于配体的荧光探针与FRAP相结合的潜力，以便未来对活细胞中CBR流动性的生化细节及其对受体依赖性细胞过程的影响进行研究。
