新生小鼠缺氧缺血期间的连续视频脑电图
Summary
本手稿描述了一种在经历缺氧缺血的新生小鼠中使用多个深度电极进行连续视频脑电图记录的方法。
Abstract
缺氧缺血是新生儿癫痫发作的最常见原因。动物模型对于了解新生儿癫痫发作和缺氧缺血的机制和生理学至关重要。本文介绍了一种在新生小鼠中连续进行视频脑电图（EEG）监测的方法，以检测癫痫发作并分析缺氧缺血期间的脑电图背景。同时使用视频和脑电图可以描述癫痫发作的符号学和确认癫痫发作。该方法还允许分析实验时间段内的功率谱图和EEG背景模式趋势。在这种缺氧缺血模型中，该方法允许在损伤前进行脑电图记录，以获得规范基线，并在损伤和恢复期间进行。总监测时间受到限制，因为无法将幼崽与母亲分开超过四个小时。虽然，我们在本文中使用了缺氧缺血性癫痫发作的模型，但这种用于新生儿视频脑电图监测的方法可以应用于啮齿动物的各种疾病和癫痫发作模型。
Introduction
缺氧缺血性脑病 （HIE） 是一种每年影响 1000 名新生儿中 1.5 名新生儿的疾病，是新生儿癫痫发作的最常见原因1，2。存活下来的婴儿有患各种神经残疾的风险，如脑瘫、智力残疾和癫痫3，4，5。
动物模型在理解和研究缺氧缺血和新生儿癫痫发作的病理生理学方面起着关键作用6，7。改进的Vannucci模型用于在出生后第10天诱导缺氧缺血（HI）（p10）7，8。这个年龄段的小鼠幼崽在神经学上大致翻译为完整的足月人类新生儿9。
与该损伤模型结合使用的连续视频脑电图 （EEG） 监测可进一步了解和表征新生儿缺氧缺血性癫痫发作。以前的研究已经使用各种方法来分析啮齿动物的新生儿癫痫发作，包括视频记录，有限的脑电图记录和遥测脑电图记录10，11，12，13，14，15，16。在下面的手稿中，我们深入讨论了在缺氧缺血期间记录小鼠幼崽连续视频脑电图的过程。这种用于新生儿小鼠幼崽连续视频脑电图监测的技术可以应用于各种疾病和癫痫发作模型。
Protocol
所有动物研究均获得弗吉尼亚大学机构动物护理和使用委员会（IACUC）的批准。 1. 电极建筑/电缆建筑 使用单极绝缘不锈钢丝（0.005″裸直径，0.008″涂层）制成与母插座连接器（母插座连接器0.079）连接的电极。 使用特殊的定制电缆将动物连接到放大器。 将一个 4 针公连接器（公连接器 0.079″）连接到 4 通道单位增益阻抗匹配运算放大器（运算放大器）。将1…
Representative Results
癫痫发作符号学 新生儿缺氧缺血暴露导致小鼠全身性和局灶性癫痫发作（图1A-C）。视频脑电图记录允许将电图结果与视频行为相关联。这些行为使用先前发表的新生儿啮齿动物行为癫痫发作评分（BSS）进行评分16。除BSS外，我们还根据行为是局灶性/单侧，双侧还是混合性对事件进行分类（图1B…
Discussion
我们已经提出了一种在缺氧缺血性癫痫发作期间对新生小鼠进行连续视频脑电图监测的模型。结合脑电图进行视频分析，可表征癫痫发作的符号学特征。脑电图分析允许提取功率谱图和背景振幅分析。
正确和小心地放置电极在此方案中至关重要，因为电极放置过程中的伤害或不正确的放置会显着影响结果。在损伤前评估正常的基线脑电图活动至关重要，因为在电极放置过程?…
Divulgations
The authors have nothing to disclose.
Acknowledgements
我们承认以下资金来源：NIH NINDS – K08NS101122 （JB）， R01NS040337 （JK）， R01NS044370 （JK）， 弗吉尼亚大学医学院 （JB）。
Materials
| SURGERY | |||
| Ball Point Applicator | Metrex Research | 8300-F | i-bond applicator |
| Cranioplast (Powder/Resin) | Coltene | H00383 | Perm Reline/Power |
| I-Bond | Kulzer GmbH, Germany | ||
| LOOK Silk Suture | Surgical Specialities Corporation | SP115 | LOOK SP115 Black Braided Silk Non absorbable surgical suture |
| RS-5168 Botvin Forceps | Roboz Surgical Instrument | RS5168 | Forcep for surgery/ligation |
| RS-5138 Graefe Forceps | Roboz Surgical Instrument | RS5138 | Forcep for surgery/ligation |
| UV light for I-Bond | Blast Lite By First Media | BL778 | UV ligth for I-bond |
| Vannas Microdissecting Scissor | Roboz Surgical Instrument | RS5618 | Scissor for ligation |
| Vet Bond | 3M Vetbond | 1469SB | Vet Glue |
| HYPOXIA | |||
| Hypoxidial | Starr Life Science | ||
| Oxygen sensor | Medical Products | MiniOxI- oxygen analyzer/sensor for hypoxia rig | |
| EEG RECORDING | |||
| Female receptacle connector 0.079" | Mill-Max Manufacturing Corp | 832-10-024-10-001000 | Ordered from Digikey |
| Grass Amplifier | Natus Neurology Incorporated | Grass Product | |
| LabChart Pro | ADI Instruments | Software to run the system | |
| Male Socket Connector 0.079" | Mill-Max Manufacturing Corp | 833-43-024-20-001000 | Ordered from Digikey |
| Operational Amplifier | Texas Instruments, Dallas, TX, USA | TLC2274CD | TLC2274 Quad Low‐Noise Rail‐to Rail Operational Amplifier |
| Operational Amplifier | Texas Instruments, Dallas, TX, USA | TLC2272ACDR | TLC2274 Quad Low‐Noise Rail‐to Rail Operational Amplifier |
| Stainless Steel wire | A-M Systems | 791400 | 0.005" Bare/0.008" Coated 100 ft |
| Ultra-Flexible Wire | McMaster-Carr | 9564T1 | 36 Gauze wire of various color |
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Citer Cet Article
Wagley, P. K., Williamson, J., Skwarzynska, D., Kapur, J., Burnsed, J. Continuous Video Electroencephalogram during Hypoxia-Ischemia in Neonatal Mice. J. Vis. Exp. (160), e61346, doi:10.3791/61346 (2020).