title: Heterogeneous expression of voltage-gated potassium channels of the shaker family (Kv1) in oligodendrocyte progenitors.
abstract: Outwardly rectifying K(+) channels determine the membrane conductance and influence the proliferation rate of glial progenitor cells. To analyze the molecular identity and the functional role of K(+) channels in glial progenitors of mouse brain, expression of shaker-type Kv1 genes was studied at three levels: (1) presence of Kv1 mRNAs, (2) biosynthesis of channel proteins and (3) electrophysiological and pharmacological properties of K(+) currents. mRNA expression of Kv1.1 to Kv1.6 genes was studied by single-cell reverse transcription-mediated polymerase chain reaction (RT-PCR) using degenerate primers to amplify the six Kv1 transcripts. Most cells expressed several mRNA combinations simultaneously. In more than half of the cells, messages for Kv1.2, Kv1.5 and Kv1.6 were found, while Kv1.1, Kv1.3 and Kv1.4 were detected in only a minority of cells. In contrast, at the level of protein expression - employing immunocytochemistry with subtype-specific antibodies - Kv1. 2 and Kv1.3 were undetectable (<2%), while almost all cells expressed Kv1.4 (85%), Kv1.5 (99%) and Kv1.6 (99%). Kv1.1 was present in a minor cell population (10%). Functional contribution of Kv1 proteins to progenitor membrane conductance was determined by analyzing the voltage-dependence of K(+) current activation and inactivation as well as their current sensitivities to the subtype-preferring blockers and toxins tetraethylammonium (TEA), 4-aminopyridine (4-AP), charybdotoxin (CTX), alpha-dendrotoxin (DTX) and mast-cell degranulating peptide (MCDP). From these results, it is concluded: first, glial progenitor cells can express all transcripts of the six Kv1 genes, but do not express all proteins; second, Kv1.4, Kv1.5 and Kv1.6 proteins are most abundant and were found in the majority of cells; and third, K(+) currents flow predominantly either through heteromeric channel complexes or through homomeric Kv1.5 ion pores, but not through homomeric Kv1.4 or Kv1.6 channels.
authors.affiliation: Max Delbrück Center for Molecular Medicine, Cellular Neurosciences, Robert-Rössle-Strasse 10, 13092, Berlin-Buch, Germany.
authors.forename: K
authors.initials: K
authors.lastname: Schmidt
authors.forename: D
authors.initials: D
authors.lastname: Eulitz
authors.forename: R W
authors.initials: RW
authors.lastname: Veh
authors.forename: H
authors.initials: H
authors.lastname: Kettenmann
authors.forename: F
authors.initials: F
authors.lastname: Kirchhoff
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D018999:Charybdotoxin / Q000494:pharmacology; D051668:Delayed Rectifier Potassium Channels; D004546:Elapid Venoms / Q000494:pharmacology; D004622:Embryo, Mammalian; D051662:Kv1.1 Potassium Channel; D051663:Kv1.2 Potassium Channel; D051664:Kv1.3 Potassium Channel; D051665:Kv1.4 Potassium Channel; D051666:Kv1.5 Potassium Channel; D008564:Membrane Potentials / Q000187:drug effects; D051379:Mice; D008815:Mice, Inbred Strains; D009457:Neuroglia / Q000166:cytology / Q000502:physiology*; D009836:Oligodendroglia / Q000166:cytology / Q000502:physiology*; D010640:Phenothiazines / Q000494:pharmacology; D015221:Potassium Channels / Q000235:genetics* / Q000502:physiology; D024642:Potassium Channels, Voltage-Gated; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology / Q000502:physiology*; D019789:Tetraethylammonium / Q000494:pharmacology; D014158:Transcription, Genetic
chemical_list: C062307:10-N-methylcarbamoyl-3,7-dimethylamino-10H-phenothiazine; D051668:Delayed Rectifier Potassium Channels; D004546:Elapid Venoms; C496947:Kcna1 protein, mouse; C496970:Kcna2 protein, mouse; C496974:Kcna3 protein, mouse; C496978:Kcna4 protein, mouse; C496982:Kcna5 protein, mouse; D051663:Kv1.2 Potassium Channel; D051664:Kv1.3 Potassium Channel; D051665:Kv1.4 Potassium Channel; D051666:Kv1.5 Potassium Channel; D010640:Phenothiazines; D015221:Potassium Channels; D024642:Potassium Channels, Voltage-Gated; D018999:Charybdotoxin; D051662:Kv1.1 Potassium Channel; D019789:Tetraethylammonium; C020273:dendrotoxin