Simultaneous recording of receptor current and intraciliary Ca2+ concentration in salamander olfactory receptor cells
Simultaneous recording of receptor current and intraciliary Ca2+ concentration in salamander olfactory receptor cells
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DOI：
10.1111/j.1469-7793.2001.00637.x
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发表时间：
2001-09-15
影响因子：
5.5
通讯作者：
Matthews, HR
中科院分区：
文献类型：
--
作者：
Reisert, J;Matthews, HR
采用吸管技术结合激光荧光显微镜技术，以荧光指示剂fluo-3为荧光标记物，同时记录了气味诱导的蝾螈嗅感受器细胞电流和细胞内Ca 2+浓度。当暴露于浓度升高的气味中1 s时，吸管电流和fluo-3荧光均呈动态增加，且随时间变化趋势相似。此后，荧光信号衰减得更慢，由逐渐增加的气味浓度引起的fluo-3荧光与吸管电流的大小呈近似线性关系，延长的气味刺激引起吸管电流和纤毛内钙浓度的同步振荡，平均周期为4.5 ± 0.12 s。当刺激溶液中不加Na+时，电流和荧光信号的振荡周期比在Ringer溶液中刺激时的振荡周期延长了1.9 ± 0.2倍，这些结果支持了目前公认的嗅纤毛内Ca ~（2+）稳态的机制，并且与由长时间气味暴露诱导的振荡代表Ca 2+和环核苷酸浓度的耦合振荡的概念一致。
The suction pipette technique was combined with laser spot fluorescence microscopy to record simultaneously odour-induced current responses and intraciliary Ca2+ concentration from isolated salamander olfactory receptor cells loaded with the fluorescent Ca2+ indicator fluo-3.When exposed for 1 s to increasing odour concentrations both the suction pipette current and fluo-3 fluorescence increased dynamically, rising with a similar time course. Thereafter, the fluorescence signal decayed more slowly, outlasting the current response by 0.56 +/- 0.12 s.The fluo-3 fluorescence evoked by progressively increasing odour concentrations varied in an approximately linear manner with the magnitude of the suction pipette current.Prolonged odour stimulation evoked synchronous oscillations in both suction pipette current and intraciliary calcium concentration with a mean period of 4.5 +/- 0.3 s.When external Na+ was omitted from the stimulating solution the oscillation period for both the current and fluorescence signals was lengthened by a factor of 1.9 +/- 0.2 in comparison with the oscillation period when stimulated in Ringer solution.These results support the currently accepted mechanism for Ca2+ homeostasis within the olfactory cilia, and are consistent with the notion that the oscillations induced by prolonged odour exposure represent the coupled oscillation of Ca2+ and cyclic nucleotide concentrations.