title: Stable expression and characterization of the human brain potassium channel Kv2.1: blockade by antipsychotic agents.
abstract: We have cloned the cDNA encoding the voltage-dependent K+ channel Kv2.1 from human brain (hKv2.1). RNase protection and RT-PCR (reverse transcriptase-PCR) experiments reveal abundant Kv2.1 transcripts in human brain with virtually no expression detectable in human heart. hKv2.1 has been stably transfected into a human glioblastoma cell line, and transformed cells display large, slowly activating outward currents. The kinetics, steady-state activation and inactivation parameters, and external tetraethylammonium sensitivity were all similar to those described previously for hKv2.1 channels transiently expressed in Xenopus oocytes or other mammalian cell lines. A number of dopamine receptor antagonist/antipsychotic agents were shown to block hKv2.1. Trifluoperizine, trifluperidol and pimozide produced time-dependent blockade of hKv2.1 with IC50 values of approx. 1-2 microM. The diphenylbutylpiperidine fluspirilene was shown to be 4-5-fold more potent than the other agents tested inhibiting hKv2.1 current with an IC50 value of 297 nM. The block produced by fluspirilene was both time- and frequency-dependent. Furthermore, fluspirilene (1 microM) shifted the midpotential of the hKv2.1 steady-state inactivation curve by approx. 15 mV in the hyperpolarizing direction. These results demonstrate the usefulness of this transfection system for the pharmacological characterization of hKv2. 1. Fluspirilene proved to be a relatively potent blocker of hKv2.1 and may provide a useful starting point for the development of more potent and selective agents active against this brain K+ channel.
authors.affiliation: Hoechst Marion Roussel, Inc., Cincinnati, OH 45215, USA.
authors.forename: B
authors.initials: B
authors.lastname: Wible
authors.forename: M K
authors.initials: MK
authors.lastname: Murawsky
authors.forename: W J
authors.initials: WJ
authors.lastname: Crumb
authors.forename: D
authors.initials: D
authors.lastname: Rampe
mesh_terms: D000818:Animals; D014150:Antipsychotic Agents / Q000494:pharmacology*; D001923:Brain Chemistry / Q000502:physiology; D003001:Cloning, Molecular; D005260:Female; D005484:Fluspirilene / Q000494:pharmacology; D015870:Gene Expression / Q000502:physiology; D005909:Glioblastoma; D006801:Humans; D015640:Ion Channel Gating / Q000502:physiology; D008969:Molecular Sequence Data; D009206:Myocardium / Q000737:chemistry; D009865:Oocytes / Q000502:physiology; D018408:Patch-Clamp Techniques; D010868:Pimozide / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D026902:Potassium Channel Blockers; D015221:Potassium Channels / Q000235:genetics*; D024661:Potassium Channels, Inwardly Rectifying; D012333:RNA, Messenger / Q000032:analysis; D014268:Trifluoperazine / Q000494:pharmacology; D014272:Trifluperidol / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000737:chemistry / Q000187:drug effects / Q000502:physiology; D014981:Xenopus; D000098854:Kcnj10 Channel
chemical_list: D014150:Antipsychotic Agents; D000098854:Kcnj10 Channel; D026902:Potassium Channel Blockers; D015221:Potassium Channels; D024661:Potassium Channels, Inwardly Rectifying; D012333:RNA, Messenger; D010868:Pimozide; D014268:Trifluoperazine; D005484:Fluspirilene; D014272:Trifluperidol