Yang S M, Doi T, Asako M, Matsumoto A, Yamashita T
Department of Otolaryngology, Kansai Medical University, Moriguchi, Osaka, Japan.
Auris Nasus Larynx. 2000 Jan;27(1):15-21. doi: 10.1016/s0385-8146(99)00068-1.
We investigated membrane electrophysiological features of dissociated vestibular ganglion neurons, using a voltage-sensitive dye and a multiple site optical imaging system. The neuronal nature of the cultured vestibular ganglion cells was confirmed by positive staining with the anti-neurofilament 200 kDa antibody, using immunocytochemical methods. Optical absorption of the dye which binds to the external surface of neuron membranes increased while the cells were depolarized during perfusion with 150 mM potassium solution. The relative ratio (deltaI/I) of optical absorption change was 0.23 +/- 0.08% (means +/- S.D., n = 16). These optical responses were wavelength dependent, therefore, the optical response apparently originated from the voltage-sensitive dye. Under our experimental conditions, photodynamic damage and pharmacological effects of the dye were either absent or insignificant. We therefore concluded that optical recording is a new, practical and non-invasive method to simultaneously monitor changes in membrane potential from cultured vestibular ganglion cells. Optical recording is expected to provide further insight into mechanisms of information processing by vestibular ganglion neurons.
我们使用电压敏感染料和多部位光学成像系统,研究了离体前庭神经节神经元的膜电生理特征。采用免疫细胞化学方法,用抗神经丝200 kDa抗体进行阳性染色,证实了培养的前庭神经节细胞的神经元特性。在用150 mM钾溶液灌注期间,当细胞去极化时,与神经元膜外表面结合的染料的光吸收增加。光吸收变化的相对比率(deltaI/I)为0.23±0.08%(平均值±标准差,n = 16)。这些光学反应是波长依赖性的,因此,光学反应显然源于电压敏感染料。在我们的实验条件下,染料的光动力损伤和药理作用不存在或不显著。因此,我们得出结论,光学记录是一种新的、实用的、非侵入性的方法,可同时监测培养的前庭神经节细胞膜电位的变化。光学记录有望为深入了解前庭神经节神经元的信息处理机制提供进一步的见解。