Infrared video patch-clamp technique for spiral ganglion neurons in rat cochlear slices
Infrared video patch-clamp technique for spiral ganglion neurons in rat cochlear slices
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DOI：
10.1080/00016480802294385
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发表时间：
2009-01
影响因子：
1.4
通讯作者：
D. Zha;Ying Lin;Zhi-ming Wang;Lianjun Lu;T. Xue;Xue Gao;Yun-Qing Li;L. Qiao;Jian-Hua Qiu
中科院分区：
文献类型：
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作者：
D. Zha;Ying Lin;Zhi-ming Wang;Lianjun Lu;T. Xue;Xue Gao;Yun-Qing Li;L. Qiao;Jian-Hua Qiu
结论：耳蜗切片和红外视频膜片钳技术可用于实时观察。为进一步研究螺旋神经节神经元的电生理特性和听转导机制提供了良好的方法和平台。目的：建立与红外视频膜片钳技术相结合的离体大鼠耳蜗切片技术，探讨大鼠耳蜗外皮的电生理特性。材料与方法：SD大鼠按出生天数分为3组（0 ~ 2天、3 ~ 6天、7 ~ 14天）。快速制作SD大鼠耳蜗切片后，采用红外差干涉对比技术和膜片钳记录观察大鼠下丘脑神经网络的电生理特性，分析影响耳蜗切片质量和膜片钳记录的因素。结果：3 ~ 6日龄SD大鼠切片成功率最高，每个耳蜗可制备2 ~ 4片。保持部分颅骨与耳蜗的连接和蜗体的完整性是制作切片的关键。耳蜗轴、耳蜗叶片的位置和切片准备时间也是影响切片质量和细胞活性的重要因素。使用红外视频膜片钳技术可以很容易地发现状态良好的SGN细胞，以帮助评估密封测试过程。全细胞记录下SGN静息膜电位为- 45.6±5.3 mV (n=52)，可记录到Na+和K+电流。
Conclusion: Cochlear slice and infrared video patch-clamp techniques can be used in real-time observation. They provide a good method and platform for further study of the electrophysiological properties and auditory transduction mechanism of spiral ganglion neuron (SGN). Objective: To establish the isolated rat cochlear slice technique combined with the infrared video patch-clamp technique to explore the electrophysiological properties of the SGN. Materials and methods: SD rats were divided into three groups according to postnatal days (0–2 days, 3–6 days and 7–14 days). After quickly making SD rat cochlear slices, the electrophysiological properties of the SGN were observed using the infrared differential interference contrast technique and patch-clamp recording, and the factors that affect the cochlear slice quality and patch-clamp recording were analysed. Results: The successful slice rate was highest in 3–6-day-old SD rats and two to four slices could be prepared using each cochlea. It is crucial to maintain the connection of partial skull with the cochlea and the volute integrity when producing slices. The position of the cochlear axis and blade, and the slice preparation time were also important factors affecting slice quality and cell activity. SGN cells in good condition could easily be found using the infrared video patch-clamp technique to help the assessment of the seal test process. SGN resting membrane potential of whole-cell recording was −45.6±5.3 mV (n=52) and the currents of Na+ and K+ could be recorded.