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 (consisting 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 protocol 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).