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Functional characterisation and subcellular localisation of HCN1 channels in rabbit retinal rod photoreceptors

Gian Carlo Demontis*, Anna Moroni†, Biagio Gravante†, Claudia Altomare†, Biancamaria Longoni‡, Luigi Cervetto* and Dario DiFrancesco†§

Gating of voltage-dependent conductances in retinal photoreceptors is the first step of a process leading to the enhancement of the temporal performance of the visual system. The molecular components underlying voltage-dependent gating in rods are presently poorly defined. In the present work we have investigated the isoform composition and the functional characteristics of hyperpolarisation-activated cyclic nucleotide-gated channels (HCN) in rabbit rods. Using immunocytochemistry we show the expression in the inner segment and cell body of the isoform 1 (HCN1). Electrophysiological investigations show that hyperpolarisation-activated currents (I_h) can be measured only from the cell regions where HCN1 is expressed. Half-activation voltage (-75.0 ± 0.3 mV) and kinetics (t_1/2 of 101 ± 8 ms at -110 mV and 20 °C) of the I_h in rods are similar to those of the macroscopic current carried by homomeric rabbit HCN1 channels expressed in HEK 293 cells. The homomeric nature of HCN1 channels in rods is compatible with the observation that cAMP induces a small shift (2.3 ± 0.8 mV) in the half-activation voltage of I_h. In addition, the observation that within the physiological range of membrane potentials, cAMP does not significantly affect the gain of the current-to-voltage conversion, may reflect the need to protect the first step in the processing of visual signals from changes in cAMP turnover.

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