Gu Ben J, Avula Pavan, Wiley James S
The Florey Institute of Neuroscience and Mental Health, University of Melbourne, Parkville, VIC, Australia.
National Clinical Research Center for Aging and Medicine, Huashan Hospital, Fudan University, Shanghai, China.
Methods Mol Biol. 2020;2041:323-334. doi: 10.1007/978-1-4939-9717-6_24.
The P2X7 receptor is a classic purinoceptor/ion channel. After activated by ATP, it opens a cation selective channel, which dilates to a large pore over tens of seconds, allowing the entry of big molecules. This unique feature is often used to evaluate this receptor's function with DNA-binding dyes (MW 300-400 Da), such as ethidium bromide and Yo-Pro-1. Here we describe two-color flow cytometry based protocols for measuring P2X7 pore dilation. One is ATP-induced ethidium uptake by real-time multicolor flow cytometry for standardized and accurate quantitation, and the other is a quick whole blood assay which is particularly useful for ex vivo study.
