可逆冷却诱导 Deactivations 对行走猫的障碍记忆的皮层贡献研究
1Cerebral Systems Laboratory,University of Western Ontario, 2Graduate Program in Neuroscience,University of Western Ontario, 3Brain and Mind Institute,University of Western Ontario, 4Department of Physiology and Pharmacology,University of Western Ontario, 5Department of Psychology,University of Western Ontario
Summary
复杂的运动在自然主义的环境需要仔细的肢体协调涉及顶叶皮层的区域。下面的协议描述了可逆冷却诱导失活的使用, 以证明顶骨面积5的作用, 在记忆制导避障的步行猫。
Abstract
在复杂的、自然主义的地形上, 关于环境障碍的感官信息可以用来快速调整运动动作以躲避。例如, 在猫, 关于一个即将到来的障碍的视觉信息可以调节步避。运动适应也可以发生独立的视觉, 因为突然的触觉输入到腿部由一个预期的障碍可以修改的步的所有四条腿, 以避免。这种复杂的运动协调涉及上结构, 如顶叶皮质。该协议描述了使用可逆的, 冷却诱导的皮层失活, 以评估顶叶皮层贡献的记忆引导障碍运动在猫。小的冷却回路, 被称为 cryoloops, 是特殊的形状, 以关闭分散的兴趣区域, 以评估其贡献的公开行为。这种方法已被用来阐明的作用, 5 在记忆引导的障碍, 避免在猫的顶叶区。
Introduction
在自然的、不平坦的地形上, 关于障碍物的感官信息可以通过视觉或触觉获得, 可以迅速地改变运动以躲避。步进运动的细致协调涉及多个皮层区域1,2。例如, 电机皮层3,4和顶叶皮层5,6,7在复杂的运动任务中被牵连, 如避障。在足动物中, 避免障碍所需的阶跃调制必须伸展至前腿和后腿。如果向前运动被延迟在前腿和 hindleg 障碍间隙之间 (可能出现作为动物踏入小心地通过一个复杂, 自然主义的环境跟踪猎物), 关于在记忆中维护的障碍的信息被用来指导一旦走了, hindleg 就会越过障碍物。
实验技术旨在解除离散皮层区域可以用来研究皮层贡献的记忆引导障碍运动。冷却诱发的皮质失活提供了一种可逆的, 可靠的, 可重现的方法来评估皮质对显性行为的贡献8。Cryoloops 由不锈钢管制成, 是特定于皮层的兴趣, 确保高度选择性和离散的基因座的失活。一旦植入, 冷冻甲醇泵通过管腔的 cryoloop 冷却的皮层区域直接下方的循环到 < 20 ° c。在这个临界温度下, 突触的传递在大脑皮层的区域被抑制。这种钝化可以通过停止甲醇的流动来逆转。该方法已用于研究皮层对感官处理和行为的贡献9,10,11,12,13,14,15,16,17, 以及跳眼移动的马达控制18和内存引导障碍运动19。
该协议的目的是使用可逆冷却诱导 deactivations 评估的参与, 顶叶皮层区的运动协调在猫。特别地, 记忆引导障碍运动被检查与或没有活跃顶叶皮层。这些方法已经被用来成功地证明了顶骨面积5在记忆制导避障中的作用, 在行走 cat19。
Protocol
所有的程序都是按照国家研究委员会的《动物保护和使用实验指南》 (第八版; 2011) 和加拿大动物保护委员会关于试验动物的护理和使用指南 (1993) 进行的, 并由西安大略大学动物学委员会动物保育委员会批准。 下面的过程可以应用于实验研究皮层贡献的运动控制在步行猫。 1. 仪器 构造用于评估障碍物记忆的仪器。注: 该仪器由2.43 米长, 29 厘米?…
Representative Results
该协议已成功地用于检查顶叶皮层贡献的障碍记忆在步行猫19。在这项研究中, cryoloops 在5和7在三成人 (> 6 月龄) 雌性猫 (图 5A) 上被植入了双侧壁区。在没有冷却 (温暖、控制条件) 的情况下, 或者当5或7区被双边钝化时, 在触觉障碍记忆模式中对动物进行了评估。 该研究的结果表明, 当5区…
Discussion
所描述的范例采用冷却诱导的离散皮层区 deactivations 的 cryoloop, 以研究记忆制导障碍的运动在猫。视觉和触觉障碍记忆范例是相当简单的动物执行, 因为他们利用自然主义的运动行为, 发生在最小的努力, 当一个动物的动机是跟随一个移动的食物来源。因此, 大部分的培训阶段是专门适应动物的测试室和冷却设备。大多数动物都需要反复接触, 佩戴安全带, 并通过皮带拴住, 然后才能舒适地和自然地在…
Divulgazioni
The authors have nothing to disclose.
Acknowledgements
我们衷心感谢加拿大卫生研究院 (NSERC) 和加拿大科学与工程研究所的支持。波斯特得到了亚历山大·格雷厄姆加拿大研究生奖学金 (NSERC) 的支持。
Materials
| Camera | IDS Imaging Development Systems GmbH | Model: UI-5240CP-C-HQ | |
| Intake tubing | Restek | 25306 | Unflanged end is submerged in the methanol reservoir while the flanged end is connected to the pump |
| Pump | Fluid Metering, Inc. | Model: QG 150 | |
| Nalgene Dewar vacuum flask | Sigma-Aldrich | F9401 | |
| Teflon tubing | Ezkem | A051754 | |
| Microprobe thermometer | Physitemp | Model: BAT-12 | |
| Flanged tube end fittings | Valco Instruments Co. Inc. | CF-1BK | Assorted colours available for colour coding. Packages include the same number of washers as fittings |
| Washers | Valco Instruments Co. Inc. | CF-W1 | Extra washers |
| Flanging kit | Pro Liquid GmbH | 201553 | |
| Tubing connector | Restek | 25323 | |
| Tubing cutter | Restek | 25069 | |
| Male thermocouple connector | Omega | SMPW-T-M | Used to make cable connection to thermometer |
| Thermocouple wire | Omega | PP-T-24S | Used to make cable connection to thermometer |
| MATLAB | MathWorks | n/a |
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Citazione di questo articolo
Wong, C., Lomber, S. G. Reversible Cooling-induced Deactivations to Study Cortical Contributions to Obstacle Memory in the Walking Cat. J. Vis. Exp. (130), e56196, doi:10.3791/56196 (2017).