Université de Paris, Institut Cochin, INSERM, CNRS, Paris, France.
Methods Mol Biol. 2022;2550:179-188. doi: 10.1007/978-1-0716-2593-4_22.
Cyclic adenosine monophosphate (cAMP) is an important ubiquitous second messenger and one of the major pathways transducing the activation of G protein-coupled receptors (GPCRs). Quantifying intracellular levels of cAMP in an accurate and high-throughput manner is, therefore, of high interest to access functional responses of GPCRs. The neurohormone melatonin is selectively recognized by two GPCRs in mammals, named MT and MT. Both have an inhibitory action on intracellular cAMP levels. Here, we describe a homogeneous high-throughput-compatible methodology routinely used in our laboratory to measure cAMP levels following activation of melatonin receptors.
环磷酸腺苷 (cAMP) 是一种重要的普遍存在的第二信使,也是转导 G 蛋白偶联受体 (GPCR) 激活的主要途径之一。因此,以准确和高通量的方式定量细胞内 cAMP 水平对于评估 GPCR 的功能反应非常重要。神经激素褪黑素被哺乳动物中的两种 GPCR 选择性识别,分别命名为 MT 和 MT。两者均对细胞内 cAMP 水平具有抑制作用。在这里,我们描述了一种在我们实验室中常规使用的适用于高通量的均相测定方法,用于测量褪黑素受体激活后 cAMP 水平。