大鼠前庭系统的外科迷路切除术研究
Summary
本协议描述了大鼠的手术迷路切除术, 是研究前庭系统的有效方法。
Abstract
为了研究前庭系统或前庭代偿过程, 已经开发了一些方法来引起前庭损伤, 包括手术或化学迷路切除术和前庭神经切断术。手术迷路切除术是一种相对简单、可靠、快速的方法。本文介绍了大鼠迷路切除术的手术技术。耳后切开术在全麻下进行, 以暴露外耳道和鼓膜, 在无镫的情况下, 鼓膜和听骨被取出。镫动脉位于镫和椭圆形窗口之间, 是一种脆弱的结构, 必须保留以获得明确的手术领域。一个孔 fenestrate 的前庭是由一个2.1 毫米的钻头, 比镫。然后, 100% 乙醇注射通过这个孔和吸气几次。显微镜下细致的解剖和细致的出血控制是获得可靠结果的必要条件。手术后立即看到前庭丢失的症状, 如眼球震颤、头部倾斜和滚动运动。rotarod 或旋转椅试验可用于客观、定量地评价前庭功能。
Introduction
前庭器官对平衡和眼睛控制是必不可少的。正常的前庭功能取决于两个内耳内前庭器官的对称传入信号。前庭虚或丢失诱发头晕、眼球震颤和体位失衡。急性损伤后, 前庭功能在几天内自发恢复, 这个过程称为前庭补偿1,2。前庭补偿的静态缺陷是一个恢复的过程中与自然休息活动的不平衡的同侧前庭核。动态缺陷的前庭补偿主要实现通过感官和行为替换 (使用视觉或体感输入)3。这些过程对神经可塑性研究有吸引力4,5。
在前庭代偿过程中, 研究了前庭系统和神经可塑性的机制, 如外科和化学迷路切除术和前庭神经切断术 5, 6 ,7,8。前庭神经切断术是一种诱导完全前庭丢失的方法, 但它是一个更困难和侵入性的程序, 可能诱发脑损伤8, 9.这种方法需要更大的手术技巧和花费更多的时间比迷路切除术。包括庆大霉素、arsanilate 和丁卡因在内的化学迷路切除术更容易, 并能产生可靠的结果10, 11, 12.然而, 耳蜗也可能受损, 前庭丢失可能会随着时间的推移而发展11。此外, 化学物质对大脑的影响, 应该保留, 以准确评估, 是不清楚的。外科迷路切除术首次在1842年的动物研究中被引入15, 并在1936年16 的大鼠中首次报告。该技术已用于许多动物研究5,17,18,19。外科迷路切除术是一种特定的、可靠的、相对简单的方法。13,14此外, 术后立即看到前庭损伤的症状。在这里, 我们描述了我们的手术技术的大鼠迷路切除术。
Protocol
这项研究是按照首尔国立大学医院 (14-0148-C1A1) 的机构动物护理和使用委员会的规定进行的, 由实验室动物护理评估和鉴定协会认证。国际。 注: 对 7-8 周龄 (200/250 克) 大 (SD) 雄性大鼠进行实验。实验开始前1周, 每只动物都适应实验室的条件。这些动物被安置在温度和湿度控制的房间里, 有一个恒定的12小时:12 小时的光: 黑暗的循环, 免费获得食物和水。 1. 迷?…
Representative Results
手术的成功由行为测试证实。所有动物都表现出单侧前庭功能丧失的典型行为。自发滚轧在手术后立即明显, 是由头部的空气粉扑引起的, 或者是在早期恢复阶段 (视频 1) 轻触身体。3 d 手术后, 动物们绕着损伤的一侧移动, 偶尔会落到左边。自发眼球震颤观察在 2 d (录影 2)。在3维内观察诱发眼球震颤。 …
Discussion
这项技术是一个有效的方法, 以创造突然, 永久性, 完整的前庭功能损失。这可以用来研究前庭病理, 如前庭神经炎, 声学肿瘤和梅尼埃的疾病。许多研究已经使用这种技术来研究前庭细胞核的神经元可塑性或相关的中心进程5,17,18,19。
最关键的步骤为成功的手术是 1) 保存的 stapedial …
Declarações
The authors have nothing to disclose.
Acknowledgements
这项研究得到了韩国卫生技术 & 发展研究所 (KHIDI) 的资助, 由大韩民国卫生部 & 福利部提供资金 (赠款号: HI15C2651)。
Materials
| ASPIRATOR KB-012 | KOH BONG & CO., LTD. | KB-012 | Medical aspirator |
| Blade: #15 | Fine Science Tools | #10015-00 | Blades for #7 Scalpel Handles, #15 |
| Carbon Steel Burrs | Fine Science Tools | #19007-05 | shaft diameter: 2.3 mm, length: 44 mm, package of 10 burrs |
| Carl Zeiss Surgical GmbH | Carl Zeiss | #6627100863 | Surgical microscope |
| Dumont #3c | Fine Science Tools | #11231-20 | Standard tip 0.17 x 0.10 mm, 11 cm |
| Dumont #5SF | Fine Science Tools | #11252-00 | |
| Dumont #7B | Fine Science Tools | #11270-20 | Serrated 0.17 x 0.10 mm, 11 cm |
| Extra Fine Bonn: straight | Fine Science Tools | #14084-08 | Iris scissors, best suited for microdissection under high magnification |
| Fine Iris Scissors: straight | Fine Science Tools | #14094-11 | Made from martensitic stainless steel, combined with molybdenum and vanadium |
| Finger Loop Ear Punch | Fine Science Tools | #24212-01 | 1 mm. Provides stability and control for researchers using the numbering system |
| Hartman | Fine Science Tools | #13002-10 | Tip width: 1 cm, serrated, 10 cm |
| Short Scalpel Handle #7 Solid | Fine Science Tools | #10003-12 | #7 short, 12 cm |
| Small Vessel Cauterizer | Fine Science Tools | #18000-03 | Replacement tip, straight knife, keeps bleeding to a minimum and therefore provides a surgical field clear of clamps and hemostats |
| Strong 207S | SAESHIN | 207S | Powerful torque at low speed, available with speed or on/off foot controller |
| Suction Tubes | JEUNGDO B&P CO., LTD. | H-1927-8 | Frazier, 18 cm |
| VICRYL | ETHICON | W9570T | Synthetic absorbable sterile surgical suture |
| Weitlaner-Locktite | Fine Science Tools | #17012-13 | Maximum spread: 4.5 cm, 2 x 3 blunt teeth, 11 cm |
| Zoletil | Virbac, France | Tiletamine-zolazepam | |
| Rompun | Bayer | Xylazine | |
| Rimadyl | Pfizer | Carprofen | |
| Septra | Pfizer | Trimethoprim-sulfonamide |
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Citar este artigo
Chang, M. Y., Park, M. K., Park, S. H., Suh, M., Lee, J. H., Oh, S. H. Surgical Labyrinthectomy of the Rat to Study the Vestibular System. J. Vis. Exp. (135), e57681, doi:10.3791/57681 (2018).