β 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 β 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-α gene from chick hair cells were co-expressed with accessory β subunits (from quail cochlea) by transient transfection of human embryonic kidney 293 cells. Addition of the β 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 β. Deactivation kinetics were used to predict tuning frequencies and thus tonotopic location if hair cells were endowed with each of the expressed channels. All β-containing channels were predicted to lie within the apical (low-frequency) 30% of the epithelium, consistent with previous in situ hybridization studies. Individual slo-α exons would be found anywhere within the apical 70%, depending on the presence of β, and other alternative exons. Alternative splicing of the slo- α channel message provides intrinsic variability in gating kinetics that is expanded to a wider range of tuning by modulation with β subunits.