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Neuroscience
用于同时测量疼痛和瘙痒相关行为的切面注射模型
用于同时测量疼痛和瘙痒相关行为的切面注射模型
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
Cheek Injection Model for Simultaneous Measurement of Pain and Itch-related Behaviors

用于同时测量疼痛和瘙痒相关行为的切面注射模型

Full Text
12,793 Views
04:59 min
September 27, 2019

DOI: 10.3791/58943-v

Yu Yamanoi1,2,3,4, Hiroki Kittaka1,2,5, Makoto Tominaga1,2,3,6

1Thermal Biology Group,Exploratory Research Center on Life and Living Systems (ExCELLS), 2Division of Cell Signaling,National Institute for Physiological Sciences, 3Department of Physiological Sciences,Graduate University for Advanced Studies (SOKENDAI), 4Research Laboratory,Ikedamohando Co., Ltd., 5Department of Molecular Biomedical Sciences, College of Veterinary Medicine,North Carolina State University, 6Institute for Environmental and Gender-Specific Medicine,Juntendo University

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Summary

通常,小鼠颈部注射模型用于评估尿原引起的划伤行为。然而,该模型只提供有关瘙痒的信息,而不是疼痛的信息。在这里,在小鼠中引入了脸颊注射模型,可用于同时测量疼痛和瘙痒相关行为。

Transcript

我们的数据表明,藻原减少抓伤行为,但辣椒素对抓伤行为的影响仍不清楚。因此,同时测量每一定量的疼痛是重要的。传统方法仅为发痒或疼痛的感觉提供信息,但脸颊注射模型分别同时提供发痒和疼痛的信息。

脸颊注射可能很难在第一,因为它需要练习区分抓伤,擦拭和梳理行为。注射和行为分析的视频演示有助于掌握技术。对于面颊注射实验,在评估前一天,用剪发器剃每只鼠标脸颊上的毛皮,并在温度、湿度和声音控制的环境中将以每秒 30 到 60 帧速率的摄像机安装到三脚架上。

将四个笼子放在相机的下面,以指示的方向放置,并在每个笼子之间放置屏幕,以防止老鼠看到对方。剃须后一天,将一只鼠标放入每个录音笼中一小时,使动物适应记录条件。如果要使用局部治疗,请此时应用。

当动物们适应时,将感兴趣的测试物质装入一个0.5毫升的注射器中,每只鼠标装有29或30针。在适应期结束时,在注射前开始录像。然后将测试物质的10微升溶液在觉醒动物的剃光脸颊中。

然后将注射的小鼠返回到各自的笼子,然后继续录像30分钟。在实验结束时,将视频加载到合适的视频编辑软件程序中。每次鼠标向脸颊抬起后腿,在一次或几秒钟内划伤数次,然后将腿作为一个划痕回合放下。

老鼠使用单前腿进行擦拭,两个前腿用于梳理,但只有擦拭行为算作与疼痛相关的行为。对于颈部注射实验,在实验前一天,用剪发器剃每个老鼠脖子上的皮毛。剃须一天后,在记录笼中适应小鼠一小时,然后将10微升的感兴趣的测试物质在清醒条件下,在剃光的颈部中。

将注射的动物返回到记录笼中,并记录小鼠行为30分钟。然后计算颈部的抓伤回合和任何面部擦拭行为,正如刚刚展示的。在颈部注射模型中,组胺诱发强烈的抓伤行为,而车辆诱发轻度抓伤。

在脸颊注射模型中,辣椒素不诱发刮伤行为,但诱发擦伤行为,被认为与疼痛有关。注射到脸颊后,组胺主要诱发刮伤行为,有微弱的擦拭行为。抗组胺试剂的应用可抑制抓痒,表明组胺引起的抓痒行为是由组胺对组胺受体的反应引起的。

在这个实验中,使用痒调解器诱发刮伤,但不是擦拭行为,表明经测试的调剂起到普鲁里托根的作用,但在小鼠中不具有藻原,类似于它在人类身上的影响。这种方法允许疼痛和疼痛的感觉很容易和清楚地区分与一个程序。详细的行为分析可以通过专注于痒或疼痛的感觉来完成。

痒和疼痛行为有不同的反应。此外,这些技术并不难。脸颊注射和观察程序可以让我们的各种训练同行掌握。

我们很欣赏耶鲁大学拉莫特教授介绍的面颊注射模型。

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