Robust Mouse Pattern Electroretinograms Derived Simultaneously From Each Eye Using a Common Snout Electrode
Robust Mouse Pattern Electroretinograms Derived Simultaneously From Each Eye Using a Common Snout Electrode
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DOI：
10.1167/iovs.14-13943
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发表时间：
2014-04-01
影响因子：
4.4
通讯作者：
Porciatti, Vittorio
中科院分区：
文献类型：
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作者：
Chou, Tsung-Han;Bohorquez, Jorge;Porciatti, Vittorio
目的。我们用双眼刺激和一个普通的非角膜电极同时记录了小鼠每只眼睛的图形视网膜电(PERG)。方法：71只氯胺酮/赛拉津麻醉小鼠(C57BL/6J，4个月龄)的每只眼睛同时记录了PERG，它们来自皮下针(活动的、鼻部的、参考的、头后部的、地面的、尾根的)，反应于两个定制的发光二极管片剂以微小不同的频率(OD，0.984赫兹；OS，0.992赫兹)产生的对比度反转(0.05周期/度，>；95%对比度)。采用单通道连续采集和锁相平均(OD，369个历元，492ms；OS，372个历元，496ms)提取每只眼的独立PERG信号。结果：双眼口鼻PERG波幅高(平均25.3µV，SD 6.6)，无眼间串扰。回答是可靠的(测试-重测变异性，会话内，14%，SD7；会话间，25%，SD9；眼球间不对称，9%，SD7；会话间，13%，SD5)。视网膜神经节细胞(RGCs)是双眼鼻部PERG的主要来源，3只小鼠视神经损伤后PERG信号消失。结论：视网膜神经节细胞是RGC功能的敏感指标，在青光眼和视神经疾病的小鼠模型中得到越来越多的应用。与目前的方法相比，双目鼻子PERG在简单性和速度方面都有了实质性的改进。它还克服了角膜电极的局限性，干扰了眼睛的侵入性程序，并便于基于双眼反应比较的实验。
PURPOSE. We recorded pattern electroretinograms (PERGs) simultaneously from each eye in mice using binocular stimulation and a common noncorneal electrode.METHODS. The PERG was derived simultaneously from each eye in 71 ketamine/xylazine anesthetized mice (C57BL/6J, 4 months old) from subcutaneous needles (active, snout; reference, back of the head; ground, root of the tail) in response to contrast-reversal of gratings (0.05 cycles/deg, > 95% contrast) generated on two custom-made light-emitting diode (LED) tablets alternating at slight different frequencies (OD, 0.984 Hz; OS, 0.992 Hz). Independent PERG signals from each eye were retrieved using one channel continuous acquisition and phase-locking average (OD, 369 epochs of 492 ms; OS, 372 epochs of 496 ms). The PERG was the average of three consecutive repetitions.RESULTS. Binocular snout PERGs had high amplitude (mean, 25.3 mu V, SD 6.6) and no measurable interocular cross-talk. Responses were reliable (test-retest variability within-session, 14%, SD 7; between sessions, 25%, SD 9; interocular asymmetry within-session, 9%, SD 7; between sessions, 13%, SD 5). Retinal ganglion cells (RGCs) were the main source of the binocular snout PERG, as optic nerve crush in three mice abolished the signal.CONCLUSIONS. The PERG, a sensitive measure of RGC function, is used increasingly in mouse models of glaucoma and optic nerve disease. Compared to current methods, the binocular snout PERG represents a substantial improvement in terms of simplicity and speed. It also overcomes limitations of corneal electrodes that interfere with invasive procedures of the eye and facilitates experiments based on comparison between the responses of the two eyes.