Hyperpolarization-activated currents through Clc-2 channels in RNS-derived rods.

<p>(<b>A</b>) Traces plot normalized inward currents generated by an EGFP⁺ cell (images of the cell are shown above) in response to a hyperpolarizing voltage step from −40 to −100 mV either in Locke solution (CNTR – black trace) or in the presence of 3 mM CsCl (CsCl - red trace). Step duration was 1.2 s. Voltages are indicated close to the traces. (<b>B</b>) Traces plot normalized inward currents activated in response to a hyperpolarizing voltage step from −40 to −140 mV in Locke solution (CNTR – black trace), in the presence of 2 mM CdCl₂ (red trace) and after reverting to Locke solution (WASH, green trace). Step duration was 2 sec. Images of the recorded cell are shown above traces. (<b>C–D</b>) Sweeps plot normalized currents activated by membrane hyperpolarizations in EGFP⁺ RNS-derived cells at D20 (C) and D30 (D). Voltage steps ranged from −80 to −140 mV in 20 mV-steps, from a holding of −40 mV. Numbers close to the experimental records in A–D indicate the applied voltage. Vertical deflections in A–D are capacitance transients elicited by voltage changes. Calibration bars hold for both C and D. (<b>E</b>) Data points plot mean normalized currents, with their s.e.m., as a function of activating voltages, for D20 (red, N = 8) and D30 (green, N = 10) RNS-derived cells. For each cell the last 200 ms of the 2 s-long steps were averaged. (<b>F</b>) Real-time PCR evaluation of <i>Clcn2</i> expression in RNS-derived cells at D20 (red), and D30 (green). Bars plot s.e.m. from a single experiment performed in triplicate. Data derive from the formula 2^−ΔCt.