Chabbert C, Chambard J M, Sans A, Desmadryl G
Institut National de la Santé et de la Recherche Médicale U432, Neurobiologie et Développement du Système Vestibulaire, 34095 Montpellier cedex 5, France.
J Neurophysiol. 2001 Mar;85(3):1017-26. doi: 10.1152/jn.2001.85.3.1017.
The nature and electrophysiological properties of Ca(2+)-independent depolarization-activated potassium currents were investigated in vestibular primary neurons acutely isolated from postnatal mice using the whole cell configuration of the patch-clamp technique. Three types of currents were identified. The first current, sensitive to TEA (I(TEA)) and insensitive to 4-aminopyridine (4-AP), activated at -40 mV and exhibited slow activation (tau(ac), 38.4 +/- 7.8 ms at -30 mV, mean +/- SD). I(TEA) had a half activation potential [V(ac(1/2))] of -14.5 +/- 2.6 mV and was inactivated by up to 84.5 +/- 5.7% by 10-s conditioning prepulses with a half inactivation potential [V(inac(1/2))] of -62.4 +/- 0.2 mV. The second current, sensitive to 4-AP (maximum block around 0.5 mM) and to alpha-dendrotoxin (I(DTX)) appeared at -60 mV. Complete block of I(DTX) was achieved using either 20 nM alpha-DTX or 50 nM margatoxin. This current activated 10 times faster than I(TEA) (tau(ac), 3.5 +/- 0.8 ms at -50 mV) with V(ac(1/2)) of -51.2 +/- 0.6 mV, and inactivated only slightly compared with I(TEA) (maximum inactivation, 19.7 +/- 3.2%). The third current, also sensitive to 4-AP (maximum block at 2 mM), was selectively blocked by application of blood depressing substance (BDS-I; maximum block at 250 nM). The BDS-I-sensitive current (I(BDS-I)) activated around -60 mV. It displayed fast activation (tau(ac), 2.3 +/- 0.4 ms at -50 mV) and fast and complete voltage-dependent inactivation. I(BDS-I) had a V(ac(1/2)) of -31.3 +/- 0.4 mV and V(inac(1/2)) of -65.8 +/- 0.3 mV. It displayed faster time-dependent inactivation and recovery from inactivation than I(TEA). The three types of current were found in all the neurons investigated. Although I(TEA) was the major current, the proportion of I(DTX) and I(BDS-I) varied considerably between neurons. The ratio of the density of I(BDS-I) to that of I(DTX) ranged from 0.02 to 2.90 without correlation with the cell capacitances. In conclusion, vestibular primary neurons differ by the proportion rather than the type of the depolarization-activated potassium currents they express.
采用膜片钳技术的全细胞记录模式,对出生后小鼠急性分离的前庭初级神经元中与Ca(2+)无关的去极化激活钾电流的性质和电生理特性进行了研究。鉴定出三种类型的电流。第一种电流对TEA敏感(I(TEA)),对4-氨基吡啶(4-AP)不敏感,在-40 mV时激活,表现出缓慢激活(在-30 mV时,tau(ac)为38.4±7.8 ms,平均值±标准差)。I(TEA)的半激活电位[V(ac(1/2))]为-14.5±2.6 mV,通过10 s的预处理脉冲可使高达84.5±5.7%的电流失活,半失活电位[V(inac(1/2))]为-62.4±0.2 mV。第二种电流对4-AP敏感(在0.5 mM左右达到最大阻断),对α-树眼镜蛇毒素敏感(I(DTX)),在-60 mV时出现。使用20 nM的α-树眼镜蛇毒素或50 nM的玛格毒素可实现对I(DTX)的完全阻断。该电流的激活速度比I(TEA)快10倍(在-50 mV时,tau(ac)为3.5±0.8 ms),V(ac(1/2))为-51.2±0.6 mV,与I(TEA)相比,仅轻微失活(最大失活率为19.7±3.2%)。第三种电流也对4-AP敏感(在2 mM时达到最大阻断),通过施加降血压物质(BDS-I;在250 nM时达到最大阻断)可被选择性阻断。BDS-I敏感电流(I(BDS-I))在-60 mV左右激活。它表现出快速激活(在-50 mV时,tau(ac)为2.3±0.4 ms)和快速且完全的电压依赖性失活。I(BDS-I)的V(ac(1/2))为-31.3±0.4 mV,V(inac(1/2))为-65.8±0.3 mV。与I(TEA)相比,它表现出更快的时间依赖性失活和失活后恢复。在所研究的所有神经元中均发现了这三种类型的电流。尽管I(TEA)是主要电流,但I(DTX)和I(BDS-I)的比例在不同神经元之间差异很大。I(BDS-I)与I(DTX)的密度比在0.02至2.90之间,与细胞电容无关。总之,前庭初级神经元的差异在于它们所表达的去极化激活钾电流的比例而非类型。