Detecting CO2-Sensitive Hemichannels in Neurons in Acute Brain Slices

This protocol provides two independent methods to functionally detect the neuronal expression of CO2-sensitive hemichannels. These hemichannels (con-sisting of connexins 26 or 30) are directly gated by CO2, independent of pH changes and until recently were thought to be only expressed by glia. This pro-tocol outlines a method to change the concentration of CO2 without changing pH, using isohydric solutions and then utilizing this to detect opening and closing of functional hemichannels using whole-cell patch clamp recording and dye loading. For complete details on the use and execution of this protocol, please refer to Hill et al. (2020).