Thermodynamic properties of hyperpolarization-activated current (Ih) in a subgroup of primary sensory neurons

Abstract

Ih is a poorly selective cation current that activates upon hyperpolarization, present in various types of neurons. Our aim was to perform a detailed thermodynamic analysis of Ih gating kinetics, in order to assess putative structural changes associated with its activation and deactivation. To select dorsal root ganglia neurons that exhibit large Ih, we applied a current signature method by Petruska et al. (J Neurophysiol 84:2365-2379, 2000) and found appropriate neurons in cluster 4. Currents elicited by 3,000-ms hyperpolarizing pulses at 25 and 33 degrees C were fitted with double exponential functions, yielding time constants similar to those of HCN1. The fast activation and deactivation rates showed temperature coefficients (Q10) of 2.9 and 3.1, respectively, while Q10 of the absolute conductance was 1.3. Using the Arrhenius-Eyring formalism we computed heights of voltage-independent Gibbs free energy and entropy barriers for each rate. The free energy barriers of the fast rates were just approximately 2RT units lower than those of the corresponding slow rates (31.3 vs. 33.2RT for activation, and 24.7 vs. 25.8RT for deactivation, at 25 degrees C). Interestingly, the entropy barriers of the slow rates were negative: -15.2R units for activation and -11.9R units for deactivation, compared to 4.6 and 1.3R units, respectively, for the fast component. The equivalent gating charge (zg) (3.75 +/- 0.32, mean +/- SEM, at 25 degrees C) and half-activation potential (V1/2) (-70.0 +/- 1.3 mV at 25 degrees C) did not vary significantly with temperature.