title: beta subunits modulate alternatively spliced, large conductance, calcium-activated potassium channels of avian hair cells.
abstract: Electrical tuning confers frequency selectivity onto sensory hair cells in the auditory periphery of frogs, turtles, and chicks. The resonant frequency is determined in large part by the number and kinetics of large conductance, calcium-activated potassium (BK) channels. BK channels in hair cells are encoded by the alternatively spliced slo gene and may include an accessory beta subunit. Here we examine the origins of kinetic variability among BK channels by heterologous expression of avian cochlear slo cDNAs. Four alternatively spliced forms of the slo-alpha gene from chick hair cells were co-expressed with accessory beta subunits (from quail cochlea) by transient transfection of human embryonic kidney 293 cells. Addition of the beta subunit increased steady-state calcium affinity, raised the Hill coefficient for calcium binding, and slowed channel deactivation rates, resulting in eight functionally distinct channels. For example, a naturally occurring splice variant containing three additional exons deactivated 20-fold more slowly when combined with beta. Deactivation kinetics were used to predict tuning frequencies and thus tonotopic location if hair cells were endowed with each of the expressed channels. All beta-containing channels were predicted to lie within the apical (low-frequency) 30% of the epithelium, consistent with previous in situ hybridization studies. Individual slo-alpha exons would be found anywhere within the apical 70%, depending on the presence of beta, and other alternative exons. Alternative splicing of the slo-alpha channel message provides intrinsic variability in gating kinetics that is expanded to a wider range of tuning by modulation with beta subunits.
authors.affiliation: The Center for Hearing Sciences, Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205-2195, USA.
authors.forename: K
authors.initials: K
authors.lastname: Ramanathan
authors.forename: T H
authors.initials: TH
authors.lastname: Michael
authors.forename: P A
authors.initials: PA
authors.lastname: Fuchs
mesh_terms: D017398:Alternative Splicing / Q000502:physiology*; D000595:Amino Acid Sequence; D000818:Animals; D002118:Calcium / Q000493:pharmacokinetics*; D002460:Cell Line; D004553:Electric Conductivity; D004558:Electric Stimulation; D006198:Hair Cells, Auditory / Q000737:chemistry / Q000502:physiology*; D006801:Humans; D015640:Ion Channel Gating / Q000502:physiology*; D007668:Kidney / Q000166:cytology; D007700:Kinetics; D051038:Large-Conductance Calcium-Activated Potassium Channel beta Subunits; D051036:Large-Conductance Calcium-Activated Potassium Channels; D008969:Molecular Sequence Data; D010802:Phylogeny; D015221:Potassium Channels / Q000235:genetics* / Q000378:metabolism; D024681:Potassium Channels, Calcium-Activated; D011784:Quail
chemical_list: D051038:Large-Conductance Calcium-Activated Potassium Channel beta Subunits; D051036:Large-Conductance Calcium-Activated Potassium Channels; D015221:Potassium Channels; D024681:Potassium Channels, Calcium-Activated; D002118:Calcium