基于钙离子成像的多波长激光诱发体外小鼠螺旋神经节细胞信号转导的实验研究

In vitro study on signal transduction in mice spiral ganglion cell stimulated by multi-wavelength laser based on calcium imaging

目的探究在安全、可接受的不同波长激光信号作用下小鼠螺旋神经节细胞信号转导功能的反应。方法离体培养小鼠螺旋神经节细胞,给予特异性荧光指示剂染色标记、光学显微镜下形态观察、钙离子Fura-2荧光激发、在成像视野中选取形态结构完整的测试细胞以及固定光纤等处理,采用可见光(波长450 nm)和近红外光(波长808 nm、1065 nm)三种波长的激光信号对螺旋神经节细胞分别辐照,用钙离子成像仪对细胞内钙离子浓度进行监测。结果小鼠螺旋神经节细胞在波长450 nm激光辐照下细胞内钙离子浓度明显上升,而在其他两种波段的激光照射下,螺旋神经节细胞未呈现钙离子浓度的明显变化。螺旋神经节细胞这种反应的强弱与光纤所处位置有关,越靠近光纤口的细胞产生的钙离子浓度变化越明显,而远离光纤口的细胞,尽管发生钙离子浓度变化的次数基本一致,但每一次变化的幅度较弱。结论小鼠螺旋神经节细胞在光信号作用下具有诱发信号转导反应的可能,而且该反应具有激光波长选择性。

Objective To research the auditory nerve transduction effects under multi-wavelength pulsed laser stimulations within a safe and acceptable signal range.Methods The real-time detection of intracellular calcium concentration was adopted by specific fluorescent indicator staining based on calcium imager.The spiral ganglion cells of mice were cultured in vitro.After fluorescent indicating,morphologic observation under optical microscope,Fura-2 calcium ion fluorescence excitation,intact morphology cells selection,fixing the optical fiber,the spiral ganglion cells were irradiated by different wavelength laser,including visible light(450 nm)and near infrared light(808 nm,1065 nm).The intracellular calcium concentration was monitored by calcium ion imaging.Results When 450 nm laser stimulated spiral ganglion cells,the intracellular calcium concentration was strongly increased,however,for other wavelength laser stimulation,there was no obvious relative response.And the sensitivity expression of the nerve cells under laser was related with the location of laser fiber.Cells closer to the fiber produced more obvious changes in calcium ion concentration,while for cells farther away from the fiber,the change amplitudes were weaker although the number of changes in calcium ion concentration was consistent.Conclusion The spiral ganglion cells of mice can induce a signal transduction response under the action of laser,and the response has laser wavelength selectivity.