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J Neurophysiol 91: 1536-1544, 2004. First published December 3, 2003; doi:10.1152/jn.00630.2003
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Cloning and Expression of a Small-Conductance Ca2+-Activated K+ Channel From the Mouse Cochlea: Coexpression with {alpha}9/{alpha}10 Acetylcholine Receptors

Liping Nie1, Haitao Song1, Mei-Fang Chen2, Nipavan Chiamvimonvat3, Kirk W. Beisel4, Ebenezer N. Yamoah1 and Ana E. Vázquez1

1 Department of Otolaryngology, University of California, Davis, California 95616; 2 Department of Neurology, University of California, Davis, California 95616; 3 Division of Cardiovascular Medicine, University of California, Davis, California 95616; 4 Department of Biomedical Sciences, Creighton University, Omaha, Nebraska 68178

Submitted 2 July 2003; accepted in final form 30 November 2003

Functional interactions between ligand-gated, voltage-, and Ca2+-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 Ca2+-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 {alpha}9 and {alpha}10 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 {alpha}9/{alpha}10 receptors, acetylcholine induced an inward current followed by a robust outward current. The results indicate that SK2 and the {alpha}9/{alpha}10 acetylcholine receptors are sufficient to partly recapitulate the native hair cell efferent synaptic response.

Address for reprint requests and other correspondence: A. E. Vázquez, Department of Otolaryngology, Center for Neuroscience, University of California, Davis, 1515 Newton Court, Room 209, Davis, CA 95616 (E-mail: avazquez{at}ucdavis.edu).




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