Department of Neuroscience, University of Virginia School of Medicine, Charlottesville, Virginia, USA.
J Neurophysiol. 2012 Jul;108(1):175-86. doi: 10.1152/jn.00098.2012. Epub 2012 Apr 11.
Inner ear hair cells respond to mechanical stimuli with graded receptor potentials. These graded responses are modulated by a host of voltage-dependent currents that flow across the basolateral membrane. Here, we examine the molecular identity and the function of a class of voltage-dependent ion channels that carries the potassium-selective inward rectifier current known as I(K1). I(K1) has been identified in vestibular hair cells of various species, but its molecular composition and functional contributions remain obscure. We used quantitative RT-PCR to show that the inward rectifier gene, Kir2.1, is highly expressed in mouse utricle between embryonic day 15 and adulthood. We confirmed Kir2.1 protein expression in hair cells by immunolocalization. To examine the molecular composition of I(K1), we recorded voltage-dependent currents from type II hair cells in response to 50-ms steps from -124 to -54 in 10-mV increments. Wild-type cells had rapidly activating inward currents with reversal potentials close to the K(+) equilibrium potential and a whole-cell conductance of 4.8 ± 1.5 nS (n = 46). In utricle hair cells from Kir2.1-deficient (Kir2.1(-/-)) mice, I(K1) was absent at all stages examined. To identify the functional contribution of Kir2.1, we recorded membrane responses in current-clamp mode. Hair cells from Kir2.1(-/-) mice had significantly (P < 0.001) more depolarized resting potentials and larger, slower membrane responses than those of wild-type cells. These data suggest that Kir2.1 is required for I(K1) in type II utricle hair cells and contributes to hyperpolarized resting potentials and fast, small amplitude receptor potentials in response to current inputs, such as those evoked by hair bundle deflections.
内耳毛细胞对机械刺激产生分级受体电位。这些分级反应受跨基底外侧膜流动的多种电压依赖性电流调制。在这里,我们研究了一类电压依赖性离子通道的分子特征和功能,该通道携带已知为 I(K1) 的钾选择性内向整流电流。I(K1) 已在各种物种的前庭毛细胞中被鉴定,但它的分子组成和功能贡献仍不清楚。我们使用定量 RT-PCR 显示,内向整流基因 Kir2.1 在胚胎第 15 天至成年期的小鼠耳石中高度表达。我们通过免疫定位证实了毛细胞中 Kir2.1 蛋白的表达。为了研究 I(K1) 的分子组成,我们记录了对 50-ms 步长从 -124 到 -54 mV 的电压依赖性电流,步长为 10 mV,增加 10 mV。野生型细胞具有快速激活的内向电流,反转电位接近 K(+)平衡电位,全细胞电导为 4.8 ± 1.5 nS(n = 46)。在 Kir2.1 缺陷型(Kir2.1(-/-))小鼠的耳石毛细胞中,在所有检查的阶段均不存在 I(K1)。为了确定 Kir2.1 的功能贡献,我们在电流钳模式下记录膜反应。Kir2.1(-/-) 小鼠的毛细胞具有明显(P < 0.001)更去极化的静息电位和更大、更慢的膜反应,而野生型细胞则没有。这些数据表明,Kir2.1 是 II 型耳石毛细胞中 I(K1) 的必需,并且有助于对电流输入(如毛束偏斜引起的电流输入)产生超极化的静息电位和快速、小幅度的受体电位。