Cloning and Expression of a Small-Conductance Ca2+-Activated K+ Channel from the Mouse Cochlea

Coexpression with α9/α10 Acetylcholine Receptors

Liping Nie, Haitao Song, Mei Fang Chen, Nipavan Chiamvimonvat, Kirk Beisel, Ebenezer N. Yamoah, Ana E. Vázquez

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Functional interactions between ligand-gated, voltage-, and Ca 2+-activated ion channels are essential to the properties of excitable cells and thus to the working of the nervous system. The outer hair cells in the mammalian cochlea receive efferent inputs from the brain stem through cholinergic nerve fibers that form synapses at their base. The acetylcholine released from these efferent fibers activates fast inhibitory postsynaptic currents mediated, to some extent, by small-conductance Ca 2+-activated K+ channels (SK) that had not been cloned. Here we report the cloning, characterization, and expression of a complete SK2 cDNA from the mouse cochlea. The cDNAs of the mouse cochlea α9 and α10 acetylcholine receptors were also obtained, sequenced, and coexpressed with the SK2 channels. Human cultured cell lines transfected with SK2 yielded Ca2+-sensitive K+ current that was blocked by dequalinium chloride and apamin, known blockers of SK channels. Xenopus oocytes injected with SK2 in vitro transcribed RNA, under conditions where only outward K+ currents could be recorded, expressed an outward current that was sensitive to EGTA, dequalinium chloride, and apamin. In HEK-293 cells cotransfected with cochlear SK2 plus α9/α10 receptors, acetylcholine induced an inward current followed by a robust outward current. The results indicate that SK2 and the α9/α10 acetylcholine receptors are sufficient to partly recapitulate the native hair cell efferent synaptic response.

Original languageEnglish
Pages (from-to)1536-1544
Number of pages9
JournalJournal of Neurophysiology
Volume91
Issue number4
DOIs
StatePublished - Apr 2004

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Calcium-Activated Potassium Channels
Cochlea
Cholinergic Receptors
Organism Cloning
Dequalinium
Apamin
Outer Auditory Hair Cells
Complementary DNA
Cholinergic Fibers
Inhibitory Postsynaptic Potentials
HEK293 Cells
Egtazic Acid
Xenopus
Ion Channels
Nerve Fibers
Synapses
Nervous System
Brain Stem
Acetylcholine
Oocytes

All Science Journal Classification (ASJC) codes

  • Physiology
  • Neuroscience(all)

Cite this

Cloning and Expression of a Small-Conductance Ca2+-Activated K+ Channel from the Mouse Cochlea : Coexpression with α9/α10 Acetylcholine Receptors. / Nie, Liping; Song, Haitao; Chen, Mei Fang; Chiamvimonvat, Nipavan; Beisel, Kirk; Yamoah, Ebenezer N.; Vázquez, Ana E.

In: Journal of Neurophysiology, Vol. 91, No. 4, 04.2004, p. 1536-1544.

Research output: Contribution to journalArticle

Nie, Liping ; Song, Haitao ; Chen, Mei Fang ; Chiamvimonvat, Nipavan ; Beisel, Kirk ; Yamoah, Ebenezer N. ; Vázquez, Ana E. / Cloning and Expression of a Small-Conductance Ca2+-Activated K+ Channel from the Mouse Cochlea : Coexpression with α9/α10 Acetylcholine Receptors. In: Journal of Neurophysiology. 2004 ; Vol. 91, No. 4. pp. 1536-1544.
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