The mammalian family of two-pore domain K+ (K2P) channel proteins are encoded by 15 KCNK genes and subdivided into six subfamilies on the basis of sequence similarities: TWIK, TREK, TASK, TALK, THIK, and TRESK. K2P channels are expressed in cells throughout the body and have been implicated in diverse cellular functions including maintenance of the resting potential and regulation of excitability, sensory transduction, ion transport, and cell volume regulation, as well as metabolic regulation and apoptosis. In recent years K2P channel isoforms have been identified as important targets of several widely employed drugs, including: general anesthetics, local anesthetics, neuroprotectants, and anti-depressants. An important goal of future studies will be to identify the basis of drug actions and channel isoform selectivity. This goal will be facilitated by characterization of native K2P channel isoforms, their pharmacological properties and tissue-specific expression patterns. To this end the present review examines the biophysical, pharmacological, and functional characteristics of cloned mammalian K2P channels and compares this information with the limited data available for native K2P channels in order to determine criteria which may be useful in identifying ionic currents mediated by native channel isoforms and investigating their pharmacological and functional characteristics.

No abstract

No abstract

Multiple genes of the TASK subfamily of two-pore domain K(+) channels are reported to be expressed in rat glomerulosa cells. To determine which TASK isoforms contribute to native leak channels controlling resting membrane potential, patch-clamp studies were performed to identify biophysical and pharmacological characteristics of macroscopic and unitary K(+) currents diagnostic of recombinant TASK channel isoforms. Results indicate K(+) conductance (gK(+)) is mediated almost exclusively by a weakly voltage-dependent (leak) K(+) channel closely resembling TASK-3. Leak channels exhibited a unitary conductance approximating that expected for TASK-3 under the recording conditions employed, brief mean open times and a voltage-dependent open probability. Extracellular H(+) induced voltage-independent inhibition of gK(+), exhibiting an IC(50) of 56 nM: (pH 7.25) and a Hill coefficient of 0.75. Protons inhibited leak channel open probability (P(o)) by promoting a long-lived closed state (tau > 500 ms). Extracellular Zn(2+) mimicked the effects of H(+); inhibition of gK(+) exhibited an IC(50) of 41 microM: with a Hill coefficient of 1.26, inhibiting channel gating by promoting a long-lived closed state. Ruthenium red (5 microM: ) inhibited gK(+) by 75.6% at 0 mV. Extracellular Mg(2+) induced voltage-dependent block of gK(+), inhibiting unitary current amplitude without affecting mean open time. Bupivacaine induced voltage-dependent block of gK(+), exhibiting IC(50) values of 116 microM: at -100 mV and 28 microM: at 40 mV with Hill coefficients of 1 at both potentials. Halothane induced a voltage-independent stimulation of gK(+) primarily by decreasing the leak channel closed-state dwell time.

The effects of the protein synthesis inhibitors anisomycin and puromycin were measured on protein synthesis and phase shifting of the circadian rhythm in the isolated Aplysia eye. Anisomycin pulses induce phase delays proportional in magnitude to the duration and percentage of protein synthesis inhibition. The phase-response curve to anisomycin pulses consisted of delays induced throughout the subjective night. Delays were maximal between circadian times (CT) 18 and CT 2; pulses initiated between CT 2 and CT 12 did not phase shift. Puromycin induced phase delays and advances. Delays were proportional to the duration and percentage of protein synthesis inhibition, occurring with increasing magnitude throughout the subjective night (CT 12-2). Peptidyl-puromycin formation may contribute to the magnitude of the delay. Advances, occurring between CT 2 and CT 8, required a greater drug concentration and pulse duration than delays and appeared to result from an effect other than protein synthesis inhibition. Our results support the hypothesis of a phase-dependent requirement for protein synthesis during the subjective night in this circadian clock.