Voltage-clamp traces of oocytes injected with
CLC-1 cRNA (B), CLC-2 cRNA (C and
D), and coinjected with both RNAs (E and
F). (A) Voltage-clamp pulse protocol. Test
pulses were applied in −20 mV steps starting at +40 mV (holding
potential: −30 mV). The tail pulse was varied depending on the cRNA
injected. Solid line, CLC-2; dashed line, CLC-1; dotted line,
coinjection. (B) Typical current traces of an oocyte
expressing CLC-1, showing the characteristic inward rectification at
positive voltages and the deactivation at negative voltages.
(C and D) Typical currents of an oocyte
expressing CLC-2. Long pulses (C) elicit a slowly activating
current at negative voltages, which deactivates relatively fast when
stepping to +40 mV. Short pulses (as in B, E, and
F) does not significantly activate CLC-2 (D).
(E and F) Typical currents of an oocyte
coinjected with CLC-1 and CLC-2 cRNA at a 1:1 concentration ratio.
Treatment with 0.2 mM 9-AC (F) abolishes a remaining
deactivating component at negative voltages, as well as the gating seen
in the tail current at a potential of −140 mV. This is more obvious
when oocytes injected with an about 10-fold excess of CLC-1 over CLC-2
cRNA (G) are treated with 9-AC (H).