Recording Hyperpolarization-Activated Cyclic Nucleotide-Gated Channel Currents (I_h) in Neurons

Abstract

Hyperpolarization-activated cyclic nucleotide–gated (HCN) channels are voltage-gated ion channels that play a crucial role in many physiological processes such as memory formation and spatial navigation. Alterations in expression and function of HCN channels have also been associated with multiple disorders including epilepsy, neuropathic pain, and anxiety/depression. Interestingly, neuronal HCN currents (I_h) have diverse biophysical properties in different neurons. This is likely to be in part caused by the heterogeneity of the HCN subunits expressed in neurons. This variation in biophysical characteristics is likely to influence how I_h affects neuronal activity. Thus, it is important to record I_h directly from individual neurons. This protocol describes voltage-clamp methods that can be used to record neuronal I_h under whole-cell voltage-clamp conditions, in cell-attached mode, or with outside-out patches. The information obtained using this approach can be used in combination with other techniques such as computational modeling to determine the significance of I_h for neuronal function.