Tomaru Akihiro, Kurahashi Takashi
Department of Biophysical Engineering, Graduate School of Engineering Science, Osaka University, Toyonaka, Osaka 560-8531, Japan.
J Neurophysiol. 2005 Apr;93(4):1880-8. doi: 10.1152/jn.00303.2004. Epub 2004 Nov 17.
Spike discharges of single olfactory receptor cells (ORCs) were recorded with the whole cell patch-clamp method applied to slice preparation. In parallel, activities of transduction channels were measured under the voltage-clamp condition. When cells were stimulated by odorants, 54 out of 306 cells exhibited inward current responses (10 mM cineole in the puffer pipette). The amplitude of the inward current was dependent on the stimulus period, reflecting the time integration for the stimulus dose, and the relation could be fitted by the Hill equation. Under the current-clamp condition, current injection induced spike discharges. In cells showing repetitive firings, the firing frequency was dependent on the amount of injected current. The relation was fitted by the Michaelis-Menten equation showing saturation. When cells were responsive to the odorant and had abilities to discharge repetitive spikes, the depolarizing responses were accompanied by repetitive spikes. In those cells, the spike frequency was dose-dependent, expressing saturation similar to the result obtained by current injection. Since both transduction channel and spike generative steps expressed saturation in their dose dependences, we explored what step(s) actually determines saturation in ORC signaling processes. By examining dose-response relations of both the current and spikes in the same cells, saturating dose was found to be dependent largely on that of the transduction step. This suggests that the dynamic range is fundamentally determined by the transduction system. In addition, a simple model derived from the nonlinearity of the plasma membrane could explain that a critical level of dynamic range was, at least in part, modified by the membrane nonlinearity.
采用全细胞膜片钳方法,在脑片制备上记录单个嗅觉受体细胞(ORC)的锋电位发放。同时,在电压钳条件下测量转导通道的活性。当细胞受到气味剂刺激时,306个细胞中有54个表现出内向电流反应(在微电极吸管中加入10 mM桉叶油素)。内向电流的幅度取决于刺激时长,反映了对刺激剂量的时间积分,这种关系可用希尔方程拟合。在电流钳条件下,电流注入可诱发锋电位发放。在表现出重复放电的细胞中,放电频率取决于注入电流的大小。这种关系可用显示饱和的米氏方程拟合。当细胞对气味剂有反应且有重复发放锋电位的能力时,去极化反应会伴随重复锋电位。在这些细胞中,锋电位频率呈剂量依赖性,表现出与电流注入结果相似的饱和现象。由于转导通道和锋电位产生步骤在剂量依赖性上均表现出饱和,我们探究了在ORC信号传导过程中究竟是哪个步骤决定了饱和现象。通过检查同一细胞中电流和锋电位的剂量反应关系,发现饱和剂量很大程度上取决于转导步骤的剂量。这表明动态范围从根本上是由转导系统决定的。此外,一个基于质膜非线性推导出来的简单模型可以解释,动态范围的临界水平至少部分是由膜非线性改变的。