Detecting CO(2)-Sensitive Hemichannels in Neurons in Acute Brain Slices

This protocol provides two independent methods to functionally detect the neuronal expression of CO(2)-sensitive hemichannels. These hemichannels (consisting of connexins 26 or 30) are directly gated by CO(2), 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 CO(2) 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).