鼠标短期和长期活动性分析视频跟踪软件
Summary
运动活性（LMA）是一个简单而容易地进行测量，在小鼠的行为。视频跟踪软件（VTS）和LMA的耦合允许的特异性和灵敏度的提高，尤其是当同手动，线相交时，LMA分析方法相比。此外，VTS允许长期跟踪鼠标LMA。
Abstract
自主活动（LMA）在老鼠及其他啮齿动物的行为是一种简单，方便地完成测量。视频目标跟踪软件（VTS）中的改进，使得它能够被连接到LMA的测试，方法与人工计分的线交叉时相比，显着提高的特异性和敏感性。此外，VTS使高通量实验。虽然类似的开放的现场测试（OFT）用于自动视频跟踪，LMA测试的独特之处在于它可以让老鼠留在他们家笼不利用导致焦虑的刺激，明亮的灯光明暗，在活跃期周期。传统上，LMA已被用于很短的时间（分钟），而较长的运动的研究（小时，天）是经常用的发射器和生物遥测植入。随着实时跟踪的选项，长，像短期LMA测试，现在可以进行录像。长期的LMA测试需要专门的但容易建造，笼，使食物和水（通常定位上笼顶）不妨碍摄像。重要的是，录像和VTS可量化的参数，如鼠标移动的路径，有困难或不可行的测量线道口和/或生物遥测。总之，LMA测试耦合到VTS中的鼠标移动，在一段较长的时间，它可以检查运动提供了一个更完整的描述。
Protocol
1。短期LMA设置和程序 短期LMA笼成立 测试短期LMA一个单一的容纳鼠标内的家笼。为了促进的VTS依赖图像分析，使用床上用品用鼠标颜色（ 即深色白老鼠和黑鼠浅色床上用品床上用品）的高对比度。 在短期LMA测试，覆盖有机玻璃笼有明确的¼11×¼英寸直径的孔，以支持适当通风。 短期LMA检测室 测试小鼠在干净的环境中，它?…
Representative Results
重复测量的结果的一个例子中，短期LMA测试图1中所示。这一数字表明，脂多糖（LPS）管理后，小鼠独立通风笼具（IVC）坐落在短期复苏的自发活动的速度比周围环境隔离罩（AEC）6安置在小鼠。虽然最初的运动活性的损失LPS注射组之间是相似的，恢复至基底LMA更迅速的下腔静脉安置老鼠的移动速度（ 图1b）和持续时间的移动（ 图1c）。重复测量模型的…
Discussion
小鼠疾病的症状包括嗜睡，利息损失在社会和环境方面的环境，全身乏力和厌食2-3。在小鼠和其他啮齿类动物，LMA可以被用来评估疾病的免疫激活后4-5。此外，长期LMA提供能够跟踪运动超过24小时（或更大）期间，允许分析治疗引起的昼夜行为或干扰睡眠/唤醒模式的改变。 LMA提供几个优势超过其他疾病的测试，因为它是无创，简单地进行重复测量或连续的方式，是微创混杂。与?…
Offenlegungen
The authors have nothing to disclose.
Acknowledgements
技术支持：支持这项研究是由美国国立卫生研究院（DK064862，NS058525和AA019357到GGF）。
Materials
| Name of Reagent/Material | Company | Catalog Number | Comments |
| Video camera | Sony | HDR-XR500V | Other, similar video cameras may work as well. The file storage format (codec/container) must be compatible with the VTS used. It is recommended when initially establishing a VTS to choose the VTS first and work with the vendor in choosing an appropriate camera, especially with respects to file format and video resolution compatibility. |
| Opaque cage dividers | Custom fabricated based on specific cage dimensions. To facilitate image analysis, dividers should be of a similar color to the cage bedding. Structurally fixed dividers are preferrable because they speed setup between multi-mice experiments, such as those involving repeated measurements. Sturdy transportable cage dividers can be constructed of 1/2 inch opaque plastic and joined via a series of cross laps. | ||
| Light or Dark bedding (opposite of mouse coat color) | Available from several laboratory animal supply distributors, depending on location. | ||
| Clear Plexiglas cage tops | Custom fabricated based on cage dimensions. | ||
| Automated video tracking software (EthoVision XT 7) | Noldus Information Technology | A variety of VTS manufacturers exist, use of other software may suffice. | |
| PC | Dell Inc. | See VTS software providers minimum PC .requirements | |
| White noise generator | Dohm | DS | A variety of white noise generators are available, depending on need. |
| Brooder clamp lights with red light bulbs | Several sizes and styles available. Number required depends on room size/lighting needs. Do not exceed 125 lumens in the testing room. | ||
| Lab bench or butcher paper | Paper should match or be very similar in color to the cage bedding. |
Referenzen
- Goulding, E. H., Schenk, A. K., Juneja, P., MacKay, A. W., Wade, J. M., Tecott, L. H. A robust automated system elucidates mouse home cage behavioral structure. Proc. Natl. Acad. Sci. U.S.A. 105, 20575-20582 (2008).
- Kent, S., Bluthé, R. M., Kelley, K. W., Dantzer, R. Sickness behavior as a new target for drug development. Trends Pharmacol. Sci. 13, 24-28 (1992).
- Dantzer, R. Cytokine, sickness behavior, and depression. Immunol. Allergy Clin. North Am. 29, 247-264 (2009).
- Godbout, J. P., et al. Exaggerated neuroinflammation and sickness behavior in aged mice following activation of the peripheral innate immune system. FASEB J. 19, 1329-1331 (2005).
- York, J. M., et al. The biobehavioral and neuroimmune impact of low-dose ionizing radiation. Brain Behav. Immun. 26, 218-227 (2012).
- York, J. M., et al. Individually ventilated cages cause chronic low-grade hypoxia impacting mice hematologically and behaviorally. Brain Behav. Immun. 26, 951-958 (2012).
- Jennings, M., et al. Refining rodent husbandry: the mouse. Report of the Rodent Refinement Working Party. Lab. Anim. 32, 233-259 (1998).
- York, J. M., Blevins, N. A., Baynard, T., Freund, G. G. Mouse testing methods in psychoneuroimmunology. Methods in Molecular Biology. 934, 243-276 (2012).
- Dantzer, R., O’Connor, J. C., Freund, G. G., Johnson, R. W., Kelley, K. W. From inflammation to sickness and depression: when the immune system subjugates the brain. Nat. Rev. Neurosci. 9, 46-56 (2008).
- Crawley, J. N. Behavioral phenotyping of rodents. Comp. Med. 53, 140-146 (2003).
- Karelina, K., et al. Social isolation alters neuroinflammatory response to stroke. Proc. Natl. Acad. Sci. U.S.A. 106, 5895-5900 (2009).
Diesen Artikel zitieren
York, J. M., Blevins, N. A., McNeil, L. K., Freund, G. G. Mouse Short- and Long-term Locomotor Activity Analyzed by Video Tracking Software. J. Vis. Exp. (76), e50252, doi:10.3791/50252 (2013).