The patch clamp technique was used to investigate the action of the anthelmintic drug, oxantel, on nicotinic acetylcholine receptor (nAChR) currents recorded from vesicles of the somatic muscle cells of the nematode parasite Ascaris suum. The amplitudes of the currents were analysed at different membrane potentials to determine the single channel conductance. Also the open and closed durations were measured to determine the kinetic properties of the activated channel. Oxantel activated single nAChR currents throughout a concentration range 10–100 μM, these currents were not observed with oxantel-free pipette solutions. The mean open time of the activated channels at a membrane potential of –75 mV and a concentration of 10 μM was 1·34 ms. At higher concentrations the open times were shorter and voltage sensitive, decreasing in duration on hyperpolarization, thus suggesting open channel block. The kinetics were analysed using a simple channel block model. The forward block rate, K + B, increased with increasing oxantel concentration but showed little increase as the membrane was hyperpolarized. K + B was 2·41×107 M−1s−1 – 50 mV and 2·64 × 107 M−1s−1 at – 100mV. The unblocking rate constant, K – B, did exhibit voltage sensitivity being 443·6 s−1 at – 50 mV and 86·8 s−1 at –100 mV. Thus the blocking dissociation constant KB (= K – B/K + B) was 18·5 μM at –50 mV and 3·3 μM at –100 mV. The simple channel block scheme was found to be insufficient to explain fully the observations made; reasons for this are discussed.

Background and objective The γ1-aminobutyric acidA receptor (GABAAR) is a target for anaesthetic agents. We investigated the interactions of sevoflurane with a recombinant GABAAR. Emphasis was on the mechanism of block, as relevant open-channel block by a volatile anaesthetic would possibly explain prolonged GABAergic postsynaptic currents.

Methods The effect of sevoflurane on GABA–induced currents through recombinant α1β1γ1 GABAAR channels was studied (patch clamp; HEK293 cells). GABA 0.01mM or 1mM was applied alone or together with sevoflurane (0.05mM to 5mM).

Results Currents elicited by GABA 0.01mM were increased by low sevoflurane concentrations to 183% and decreased by high sevoflurane concentrations (> 1mM) to 34% (P < 0.05). Ten- to 90%-rise times of the currents were reduced by sevoflurane concentration dependently. At GABA (1mM), peak currents and 10–90%-rise times decreased with increasing sevoflurane concentrations. A transient current increase was induced by discontinuation of GABA and sevoflurane. Such rebound currents indicate a reversal of an openchannel block by sevoflurane.

Conclusions Sevoflurane (a) increases the apparent affinity of GABA to the GABAAR, as suggested by the decreased current rise times. This explains the enhancement of the currents induced by low GABA concentrations (0.01mM). Additionally, sevoflurane (b) induces a picrotoxin-like open-channel block at the GABAAR. The reversal of the open-channel block elicits a delayed GABA response. These findings indicate at least two different sites of action of sevoflurane at this receptor that are both important for an enhanced GABAergic synaptic transmission.

In switching from studying native cyclic nucleotide-gated (CNG) ion channels in rod cells to studying the corresponding cloned channels expressed in Xenopus oocytes, we changed our perfusion system to a more efficient one. This change involved replacing culture flasks and a small plexiglass/glass chamber with plastic syringes, metal needles, and plastic petri dishes. We now report that these new perfusion system components release agents that distort or obscure measured functional properties of rod CNG channels. The magnitude and time course of appearance of the artifacts vary widely among individual components (e.g. from syringe to syringe). The effects most resemble voltage-dependent block of the channels, giving a decrease in current at positive potentials, and producing distortions of the kinetics and voltage dependence of channel activation.