Takeuchi Hiroko, Kurahashi Takashi
Department of Biophysical Engineering, Osaka University, Japan.
J Physiol. 2002 Jun 15;541(Pt 3):825-33. doi: 10.1113/jphysiol.2002.016600.
The effects of cyclic nucleotide monophosphate (cNMP) in the ciliary cytoplasm of the olfactory receptor cell were examined by using photolysis of caged cNMP loaded from the whole-cell patch clamp pipette. Illumination of the cilia induced an inward current at -50 mV. The current amplitude was voltage dependent and the polarity was reversed at +10 mV. The amplitude of the light-induced current was dependent on both light intensity and duration. The intensity-response relation was fitted well by the Hill equation with a coefficient (n(H)) of 4.99 +/- 2.66 (mean +/- S.D., n = 19) and the duration-response relation with a coefficient of 4.03 +/- 1.43 (n = 17). The activation time course of adenylyl cyclase was estimated by comparing the light-induced response with the odorant-induced response. Adenylyl cyclase was activated approximately 260 ms later from the onset of the odorant-stimulation. The light-induced current developed very sharply. This could be explained by the sequential openings of cAMP-gated and Ca2+-activated Cl- channels. At +100 mV, where Ca2+ influx is expected to be very small, the current rising phase became less steep. When the cells were stimulated by long steps of either odour or light, the odorant-induced current showed stronger decay than the light-induced response. This observation suggests that the molecular system regulating desensitization is situated upstream of cAMP production.
利用从全细胞膜片钳吸管加载的笼状环核苷酸单磷酸(cNMP)的光解作用,研究了嗅觉受体细胞纤毛细胞质中cNMP的作用。纤毛光照在-50 mV时诱导内向电流。电流幅度与电压有关,在+10 mV时极性反转。光诱导电流的幅度取决于光强度和持续时间。强度-反应关系用希尔方程拟合良好,系数(n(H))为4.99±2.66(平均值±标准差,n = 19),持续时间-反应关系系数为4.03±1.43(n = 17)。通过比较光诱导反应和气味剂诱导反应来估计腺苷酸环化酶的激活时间进程。气味剂刺激开始后约260毫秒,腺苷酸环化酶被激活。光诱导电流发展非常迅速。这可以通过环磷酸腺苷门控和钙离子激活的氯离子通道的顺序开放来解释。在+100 mV时,预计钙离子内流非常小,电流上升阶段变得不那么陡峭。当细胞受到长时间的气味或光刺激时,气味剂诱导的电流比光诱导的反应衰减更强。这一观察结果表明,调节脱敏的分子系统位于环磷酸腺苷产生的上游。