Impedance analysis of GPCR-mediated changes in endothelial barrier function: overview and fundamental considerations for stable and reproducible measurements

摘要: The past 20 years has seen significant growth in using impedance-based assays to understand the molecular underpinning of endothelial and epithelial barrier function response physiological agonists pharmacological toxicological compounds. Most studies on use G protein-coupled receptor (GPCR) which couple fast transient changes properties. power techniques such as electric cell-substrate impedance sensing (ECIS) resides its ability detect minute cell layer integrity label-free real-time ranging from seconds days. We provide a comprehensive overview biophysical principles, applications, recent developments methodologies. Despite extensive application analysis research, little attention been paid data critical experimental variables, are both essential for signal stability reproducibility. describe rationale behind common ECIS presentation interpretation illustrate practical guidelines improve intensity by adapting technical parameters electrode layout, monitoring frequency, or parameter (resistance versus magnitude). Moreover, we discuss impact parameters, including source, liquid handling, agonist preparation kinetics. Our discussions supported obtained human microvascular cells challenged with three GPCR agonists, thrombin, histamine, sphingosine-1-phosphate.