Name: a rat model of central fatigue using a modified multiple platform method
Uploaded: 2018-08-14T13:30:00.000+00:00
Duration: 5 min 13 s
Description: Watch this Scientific Journal Video about A Rat Model of Central Fatigue Using a Modified Multiple Platform Method at JoVE.com

这项工作得到北京自然科学基金 (7162124 号) 和北京中医药大学新奥基金会的支持。

所有的动物都是按照中国关于实验室动物的道德使用和护理的法律来维护的。
1. 预建模前准备
<ol><li>
实验室准备
	<ol>
<li>在实验前至少30分钟运行紫外线灯。</li>
		<li>控制实验室温度在 25 3 摄氏度, 相对湿度约30% 左右。</li>
		<li>打开实验室灯在6:00 和关闭在18:00 建立一个 12 h/12 h 光/暗循环。</li>
	</ol>
</li>
	<li>
多平台箱结构
	<ol>
<li>构造一个不透明的塑料罐, 不盖 110 x 60 x 40 厘米3。</li>
		<li>固定十五个圆形平台 (h = 8 cm, d = 6.5 cm) 在坦克的底部, 有序地分布在三行和五列。在每个平台之间留出足够的空间, 各列之间大约有10厘米, 行之间有13厘米。</li>
		<li>在水箱的侧面设置一个出水口, 并安装一个水龙头。</li>
		<li>用一个挂在上面的食物盒做一个铁网盖。</li>
	</ol>
</li>
	<li>
分组和住房大鼠 注: 实验中使用8周大的雄性大鼠, 重约 200-210 克。在建模过程中, 老鼠成群地活着。<ol>
<li>用记号笔数老鼠的尾巴根。</li>
		<li>对大鼠进行称重, 排除极轻或重的老鼠, 将其余的随机分为模型和对照组。</li>
		<li>把老鼠轻轻地放进干净的笼子里, 让他们适应实验室至少3天。提供充足的水和食物供应。</li>
	</ol>
</li>
</ol>2. 用 MMPM 建模
注: 该过程从18:00 开始, 第二天以8:00 为单位, 每天总计14小时, 超过21天。为了避免干扰因素, 同一个人需要进行整个实验, 同时穿着相同的实验室大衣。实验中使用了10只大鼠。
<ol><li>把油箱放在平坦的表面,例如地板上。然后用大约7厘米的温暖 (25 3 °c) 的水填满油箱, 在平台平坦的下面大约1厘米。</li>
	<li>准备足够的食物和饮料为所有的老鼠在坦克1天。把饲料和水放进食物盒里, 把它挂在盖子上。 注意: 一些聪明的老鼠学会在食物盒上休息。如果是的话, 把他们送回坦克。</li>
	<li>把模型组的老鼠从笼子里拿出来, 抓住它们的尾巴, 然后轻轻地放进油箱里。在水中发射所有的老鼠, 而不是平台来激发他们对水的恐惧。确保每只老鼠都有一个平台站立, 而控制组的老鼠留在他们原来的笼子里, 有足够的食物和水。</li>
	<li>把油箱盖上监视老鼠以避免意外伤害。如果一只老鼠在水中停留超过1小时而不爬上平台, 从油箱中取出, 然后从测试中取下。</li>
	<li>14小时后, 把模型鼠从水箱里拿出来, 用烘干机擦干头发。如果老鼠的尾巴褪色, 就重新标记。把老鼠送回原来的笼子里, 给他们提供足够的食物和水。</li>
	<li>冲洗油箱的每个角落。抬高油箱一侧, 打开水龙头流出污水。</li>
	<li>用75% 乙醇喷雾消毒油箱, 并将其暴露在紫外线照射下。</li>
</ol>3. 模型评估: 行为测试
注意: 所有的测试都是在行为实验室进行的。在测试期间不允许噪音和额外光线, 以避免干扰。如果可能, 请使用同一个人进行每次测试。图像处理中灰度识别需要深色外套和手套。执行经常第一, 因为它对老鼠的行为影响最小。
<ol><li>
经常
	<ol>
<li>在打开的字段框上检查录音机, 确保它正确地连接到工作站上, 并覆盖盒子的每个角落。调整照明以消除框中的阴影。</li>
		<li>把老鼠搬进他们原来的笼子里的行为实验室。允许他们在测试前至少适应1小时。</li>
		<li>用75% 乙醇清洁和消毒盒子, 以确保以前的实验没有留下任何粪便或气味。</li>
		<li>从笼子里取出一只老鼠的背部, 轻轻地放进盒子的中央区域。迅速撤退的武器从箱子, 以免阻止射击。</li>
		<li>输入老鼠的号码, 开始录音。计数和记录大鼠垂直活动的频率, 包括饲养和攀登。</li>
		<li>5分钟后, 停止录音, 把老鼠从盒子里拿出来, 把它还给笼子。</li>
		<li>重复步骤 3.1.3 3.1.6 直到所有的老鼠都完成了测试。</li>
	</ol>
</li>
	<li>
EPM
	<ol>
<li>对经常 (步骤 3.1.1 3.1.2) 执行预检和驯化步骤。</li>
		<li>把老鼠从笼子里拿回来, 轻轻地放到两只胳膊的接合处。把老鼠朝左张开的手臂, 并迅速离开, 以免阻止射击。</li>
		<li>输入老鼠的号码, 开始录音。计数和记录不同臂入口的频率。如果老鼠在测试中掉下迷宫, 把它捡起来送回迷宫。记录数据分析的详细信息。</li>
		<li>5分钟后, 停止录音, 把老鼠拿出来, 把它还给笼子。</li>
		<li>去除粪便, 用75% 乙醇擦拭迷宫, 消除前鼠的气味。</li>
		<li>重复步骤 3.2.2 3.2.5 直到所有的老鼠都完成了测试。</li>
	</ol>
</li>
	<li>
ES 试验
	<ol>
<li>填充泳池 (70 x 30 x 110 厘米3) 与 80 cm 温暖 (25 @ 3 °c) 水。 注: 如果水箱中有恒温器, 水温应设定在37摄氏度左右, 这与大鼠的体温相似。如果没有, 将其设置为室温以保持其恒定。</li>
		<li>给每只有针束的老鼠装上重物, 然后轻轻地把它绑在尾巴根部。负荷重于大鼠体重的10%。</li>
		<li>抓住一只老鼠的尾巴, 把它扔进泳池。如果老鼠蜷缩或附着在墙上, 把它们分开, 把它们驱回水中。</li>
		<li>开始计时的时候, 当老鼠被放进水中, 停止时间时, 它的疲惫, 这是证明是失败的水与嘴和鼻子在水下超过十年代。 注意: 有时, 疲惫和溺水突然发生。一定要有足够的实验者记录和保存的动物在同一时间。</li>
		<li>把筋疲力尽的老鼠从水中取出, 不要打断别人。干他们的头发, 重新标记他们的数字, 并把他们送回笼子。</li>
		<li>一组完成后, 更换水箱中的水。在所有的老鼠都完成后, 清空泳池, 用乙醇和紫外光清洁和消毒。</li>
	</ol>
</li>
</ol>4. 模型评估: 中枢神经递质检测
<ol><li>麻醉大鼠腹腔内注射10% 水合氯醛 (3 毫升/千克) 直到昏迷。</li>
	<li>斩首老鼠。</li>
	<li>沿后内侧线进行纵切口, 打开颅骨两侧, 露出大脑。把头骨翻过来, 把大脑移开, 把大脑放在冰袋上。</li>
	<li>分离并除去下丘脑, 它是位于大脑基底中央部分的菱形区域, 与周围组织有明显的边界。把它放在不育的管子里, 用液氮冷冻。把所有的样品都存放在-80 摄氏度的冰箱里。</li>
	<li>用高效液相色谱法 (HPLC)8测定下丘脑中 DA 和 5 HT 的含量。</li>
</ol>

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作者没有什么可透露的。

MMPM 最初设计为睡眠剥夺9。老鼠被发射到水箱里, 平台固定在底部。由于对水的本能的恐惧, 老鼠仍然站在平台上, 没有睡眠发生。研究表明, 不同时间的睡眠剥夺导致大鼠行为和情绪的变化, 包括认知障碍10, 消极情绪11, 中心疲劳。一些研究人员证明, 采用单平台法 (SPM) 的慢性睡眠剥夺可诱发中枢性疲劳, 并有认知障碍和社会障碍12。其他研究表明, 连续几天间歇性剥夺会导致情绪紊乱和中枢性疲劳, 这可以用内啡肽13治疗。我们以前的研究证明, 与5天14天相比, 21 天的剥夺导致更多的中枢性疲劳, 而不是与压力相关的情绪障碍14。MMPM 的许多因素可能导致中枢性疲劳, 包括长时间站立、狭小的遮蔽空间、单调乏味和重复的环境以及缺乏睡眠。睡眠剥夺与中枢性疲劳之间的潜在相关性可能与下丘脑-垂体-肾上腺 (HPA) 轴在不同的水平, 其中单胺类神经递质的变化可能发挥关键作用。
在本协议中, 我们开发了一个使用 MMPM 的中心疲劳模型, 并通过行为测试和神经递质检测对其进行评估。首先, 我们在实验室创建一个 12 h/12 h 光/暗循环 (6:00-18:00) 来模拟大鼠的自然昼夜节律, 其平均睡眠时间为12.6 小时, 夜间约 2.4-4.2 小时, 白天 8.2-9.6 小时.然后将模型鼠放入多个平台盒中, 每天连续21天站立14小时 (18:00-8:00), 对照组设置为对比。实验结束时, 模型组中的大鼠表现出明显的疲劳状态, 包括钝毛、微弱的尾部颜色、暗淡的眼睛以及笼中活动的减少。
行为测试的结果显示身体和情绪方面的变化。经常被广泛应用于啮齿动物模型评估中, 以评估勘探行为和自愿活动16。啮齿类动物有 thigmotaxis 的本能, 即一旦它们被放进一个开阔的领域, 它们就会迅速地移动到墙附近的外环。同时, 他们对新的环境感到好奇, 并且渴望通过垂直饲养和横向运动来探索中心区域。两种动机的冲突反映了焦虑情绪17。结果表明, 模型大鼠的自愿活动下降的平均速度下降的基础上。此外, 模型大鼠的饲养频率与控制大鼠相比明显降低, 这可能意味着焦虑情绪。此外, 模型大鼠在进入外环时往往花更多的时间, 在探索中没有明显的偏好, 提示模型大鼠的空间认知障碍。该测验是评估焦虑的经典测试。焦虑的大鼠倾向于保持密切的手臂安全, 而不是探索开放的手臂18。结果表明, 模型大鼠与对照组相比, 在闭合臂上花费的时间较多, 张开臂的时间较短, 与不同臂的进入频率相似, 总体上验证了模型大鼠的焦虑情绪。在 ES 试验中, 模型大鼠的游泳时间比控制大鼠短得多, 提示疲劳导致的肌肉表现不佳。结果表明, 模型大鼠焦虑、认知障碍、肌肉表现不佳、自愿活动有限, 均表现为中枢性疲劳。
对于中枢神经系统, 中枢神经递质的所有变化都表明中枢疲劳。我们发现大容量的显著下降和下丘脑的 5 HT 的增加。5-HT 是由氨基酸色氨酸 (跨国激进党) 合成的单胺类神经递质。剧烈的活动会增加5多羟的世代, 释放更多的自由激进党进入血液;累积的 5 HT 作为回报, 抑制了电机系统的中央控制, 导致较差的肌肉验19。DA 是一种易激动的神经递质, 在运动神经活动开始时增加, 并通过疲劳20的出现而下降。DA 和 5-HT 关联并相互作用作为一个励磁抑制系统, 影响到电机系统的中心控制21。因此, DA 与 5-HT 比值的下降是中枢疲劳的一个重要指标。
《议定书》中有一些对成功至关重要的说明。首先, 对雄性大鼠的模型持续时间和条件进行了测试。温度偏好和睡眠持续时间在菌株和性别之间差异14。其次, 大鼠应该在一个笼子里生活, 最多有6只老鼠, 在整个实验中提供足够的食物和水。在建模的头两个星期里, 老鼠很恼火, 可能在坦克和笼子里打架。继续监视他们, 防止受伤的老鼠死亡。另外, 记得在从水箱中取出后, 把老鼠的头发晒干, 尤其是在冬天, 以防寒冷的情况。
虽然该模型是为中心疲劳设计的, 但增加复杂因素以扩大其使用是可行的。例如, 我们安装振动马达和弹簧到平台上模拟海浪和改变剥夺模式, 试图建立一个导航疲劳模型。通过调整模型持续时间, 可以在许多其他字段中使用。作为动物模型研究, 该研究有其局限性。首先, 模型的预测有效性没有证据。在今后的研究中, 我们应该对大鼠进行抗疲劳治疗, 并对其恢复进行评估, 以证明模型的有效性。此外, 目前对该模型的评价更侧重于消极情绪和中枢神经系统的变化;然而, 中央疲劳也体现为学习困难和社会回避12。行为测试如莫里斯水迷宫和社会互动测试可以进行, 以获得更全面的了解疾病。我们希望在这里引入的中心疲劳模型可以帮助探讨中枢疲劳的病理机制。

疲劳是威胁人类健康的主要因素之一1。在过去的几十年里, 各种研究已经证明疲劳是外围触发的, 但集中驱动, 总是伴随着情感和认知障碍。意大利生理学家 a. Mosso 首先提出了 "中心疲劳2" 这个词。它一般被定义为有限的自愿活动和认知功能障碍, 由于中枢神经系统的冲动传输功能障碍 (CNS)3。与周围肌肉疲劳相比, 中枢性疲劳强调中枢神经系统的变化, 以及由此产生的情绪/行为紊乱, 包括抑郁、焦虑、认知障碍和记忆力减退。一项研究表明, 许多因素会导致中枢性疲劳, 其中过度的体力活动和精神压力是相当不可缺少的4。至于发病机制, 如色氨酸-犬尿氨路径假说5解释某些途径的变化;然而, 还需要更深入的研究来揭示中枢性疲劳的中枢-周边相关性。
由于中心疲劳的基本机制尚不清楚, 有效的动物模型对进一步的研究具有十分重要的意义。现有的疲劳模型大多是由过度运动引起的, 如跑步机6和负重游泳7, 很少关注心理因素。为了更好地模拟中心疲劳的发展, 我们小组开发了 MMPM 的大鼠模型。在建模过程中, 老鼠仍然站在狭窄的平台上, 在多个平台盒长时间, 包括部分睡眠时间。与过度运动模型不同, MMPM 模型采用部分睡眠剥夺作为心理因素, 考虑到复杂的中枢性疲劳发病机制。
对于模型评价, 我们使用经常和测验来确定焦虑情绪和自愿活动。采用 ES 试验对周围肌肉的性能进行测量。此外, 我们采取大鼠的大脑和检测多巴胺 (DA)/血清素 (5 HT) 的内容, 在两个 hypothalamuses 观察中枢神经递质的差异。
下面提出的协议旨在模拟在人类生活中常见的情况下, 反复的体力活动和缺乏睡眠导致的中枢疲劳。然而, 通过调整模型的持续时间, 它可以用于许多其他领域, 如睡眠观察和应力研究。在今后的研究中, 我们希望这一模型能帮助发现更多的中枢神经系统变化及其与周边体系的联系, 揭示中枢性疲劳的发病机制。

<table><tbody><tr><td>multiple platform sleep deprivation water tank</td><td>Customization,it is provided by the neuroimmunological laboratory of Beijing University of Chinese Medicine</td><td></td><td>110cm x 60cm x 40cm. There are 15 plastic small platforms at the bottom. The small platform is 6.5cm in diameter and 8cm high</td></tr><tr><td>Wistar rats</td><td>Beijing Weitong Lihua Experimental Animal Technology Company</td><td>license number SYXK (Beijing) 2016-0011</td><td>Use 32 Wistar healthy male rats ,8 week old (200-210 g)</td></tr><tr><td>Agilent 1100LC high performance liquid chromatograph&amp;nbsp;</td><td>Agilent</td><td>&amp;nbsp;G1379A, G1311A, G1313A , G1316A&amp;nbsp;</td><td>&amp;nbsp;G1379A, G1311A type chromatographic pump, G1313A automatic sampler, G1316A column temperature box</td></tr><tr><td>DECADE II SDC electrochemical detector</td><td>Dutch ANTEC company</td><td></td><td>glassy carbon electrode, Ag/AgCl reference electrode, workstations (Clarity CHS)</td></tr><tr><td>Biofuge Stratos high-speed refrigeration centrifuge</td><td>HERAEUS</td><td></td><td></td></tr><tr><td>VCX130 ultrasonic fracturing instrument</td><td>SONICS</td><td></td><td></td></tr><tr><td>ACS-ZEAS electronic scale</td><td>Phos technology development, Beijing.</td><td></td><td>The weight of the weighing rats can be accurate to 0.1g.</td></tr><tr><td>Open Field Box</td><td>Customization,it is provided by the neuroimmunological laboratory of Beijing University of Chinese Medicine</td><td></td><td>wooden box of open field&amp;nbsp; 100 cm by 100 cm x 40 cm, inside wall and bottom as the gray.The bottom is divided into 25 equal area squares, each of which is 20cm x 20cm, and the 16 grids along the outer wall are the external ones, and the other 9 grids are central.The camera is mounted above the median.</td></tr><tr><td>Elevated Plus-maze</td><td>Beijing zhongshi dechuang technology development co. LTD.</td><td></td><td>The open arms and close&amp;nbsp; arms of the cross are composed of 30cm x 5cm x 15cm, and the central area is 5cm x 5cm, with a camera mounted above the center and 45cm high.</td></tr><tr><td>rat swimming bucket.</td><td>Zhenhua biological instrument equipment co., LTD. Anhui,China.</td><td></td><td>The volume of plastic drum is 70cm x 30cm x 110cm, which is used for swimming in rats.</td></tr><tr><td>Thermometer</td><td>Shiya instrument co., LTD., changzhou,China.</td><td></td><td>Control water temperature</td></tr><tr><td>Small water pump</td><td>Xincheng technology co., LTD., chengdu,China.</td><td></td><td>Used for water tank and swimming behavior.</td></tr><tr><td>Ethovition3.0 behavioral software.</td><td>Nuldus,Netherlands</td><td></td><td>Measurement analysis of rat behavior videos.</td></tr></tbody></table>

a rat model of central fatigue using a modified multiple platform method

本文采用改进的多平台法 (MMPM), 介绍了一种大鼠中心疲劳模型。多平台箱设计成一个水箱, 底部有狭窄的平台。模型鼠被投入了坦克并且在平台上站立了14小时 (18:00-8:00) 每天连续21天, 与空白控制小组设置为对比。建模结束时, 模型组大鼠表现出明显的疲劳状态。为了评估模型, 我们进行了几个行为测试, 包括开放场测试 (常), 高附加迷宫 (吸盘) 测试, 和详尽的游泳 (ES) 测试。结果表明: 模型大鼠焦虑、空间认知功能障碍、肌肉表现不佳、自愿活动减少, 证实了中枢性疲劳的诊断。中枢神经递质的变化也验证了结果。最后, 该模型成功地模拟了中心疲劳, 未来的研究与模型可以帮助揭示疾病的病理机制。

我们用 MMPM 描述了一种大鼠中心疲劳模型。24只大鼠随机分为对照组和模型组, 每组12只大鼠。该模型装置设计成一个水箱, 底部有狭窄的平台 (图 1)。模型鼠每天在平台上站立14小时, 包括部分睡眠时间, 21 天 (图 2)。
行为测试是在建模后进行的, 以评估大鼠的情绪和物理变化。通常的结果 (图 3) 表明, 与对照组相比 (n = 10), 在模型大鼠 (n = 10) 的饲养运动和自愿活动的平均速度 (p < 0.05, p < 0.01) 显著下降, 和外环入口的延迟明显增加 (p < 0.01)。该测试 (图 4) 显示, 21 天的建模减少了张开臂输入和持续时间的频率与对照组相比 (n = 10) (p < 0.05, p < 0.01), 同时增加了接近臂进入和持续时间的频率 (p < 0.05)。ES 试验 (图 5) 的结果表明, 模型组 (n = 10) 的游泳持续时间明显少于对照组 (n = 10) (p < 0.001)。
接下来, 我们检测到两个 hypothalamuses 中的 DA 和 5 HT 含量, 观察中枢神经递质的差异。结果 (图 6) 显示, 与对照组相比 (n = 10) (p < 0.05, p < 0.01), 下丘脑的 da 和 da 到5的比值显著降低 (n = 10), 而 5 HT 含量增加显著 (p < 0.05)。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/57362/57362fig1.jpg"> 图 1: 多平台框的示意图.(A) 前视图。(B) 最高视图。多平台盒是一个 coverless 塑料罐 (110 x 60 x 40 cm3) 与十五个丙烯酸平台固定在底部和一个水龙头的侧面。每个平台由一根柱子和一个圆形平面 (d = 6.5 cm) 平台组成, 大于柱子顶部。平台 (h = 8 厘米) 分布在三行和五列中。相邻的平台在列中相隔10厘米, 行中为13厘米。该罐最多可容纳15只老鼠。<a href="/files/ftp_upload/57362/57362fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/57362/57362fig2.jpg"> 图 2: 建模的照片.站在站台上的老鼠是建模的第十五天。它的干头发和暗淡的眼睛表明一个明显的疲劳状态。
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/57362/57362fig3.jpg"> 图 3: 经常分析.(A) 垂直活动频率的比较。数据显示为均值电子扫描电镜 (n = 10)。以不等方差 (F = 9.877, p = 0.006 < 0.05), 意义由独立样品 t 测试, t = 2.226, p = 0.049 < 0.05 决定。模型大鼠 (n = 10) 与对照组相比, 垂直活动 (饲养) 的频率下降 (n = 10)。(B) 对自愿活动的平均速度进行比较。数据显示为中 IQR (n = 10)。意义是由曼-惠特尼 U 测试, z =-2.685, p = 0.007 < 0.01 决定。模型大鼠自愿活动的平均速度与对照组相比下降。(C) 对外环入口的延迟进行比较。数据显示为均值电子扫描电镜 (n = 10)。以不等方差 (F = 5.748, p = 0.028 < 0.05), 重要性由 t 测试, t =-3.724, p = 0.03 < 0.01 决定。模型大鼠的外环入口潜伏期增加, 这意味着它们比控制大鼠花更多的时间进入出环。注: p< 0.05 (*);p <0.01 (**);p < 0.001 (**)。
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/57362/57362fig4.jpg"> 图 4: 对吸盘测试的分析.(A) 对张开臂输入频率的比较。数据显示为均值电子扫描电镜 (n = 10)。具有等方差 (F = 0.982, p = 0.348 > 0.05), 意义由 t 检验, t = 2.710, p = 0.014 < 0.05 决定。模型大鼠 (n = 10) 的张开臂输入频率与对照鼠 (n = 10) 相比下降。(B) 在张开臂上的持续时间比较。数据显示为均值电子扫描电镜 (n = 10)。具有等方差 (F = 0.100, p = 0.755 > 0.05), 意义由 t 检验, t = 3.304, p = 0.004 < 0.01 决定。模型大鼠的张开臂持续时间与控制大鼠相比下降, 这意味着模型大鼠在张开臂上的时间较少。(C) 接近臂输入频率的比较。数据显示为均值电子扫描电镜 (n = 10)。具有等方差 (F = 0.141, p = 0.712 > 0.05), 意义由 t 检验, t =-2.466, p = 0.024 < 0.05 决定。模型大鼠闭合臂项的频率随对照组的增大而增加。(D) 密切武装的持续时间比较。数据显示为均值电子扫描电镜 (n = 10)。以不等方差 (F = 4.796, p = 0.042 < 0.05), 重要性由 t 测试, t =-2.736, p = 0.0016 < 0.05 决定。模型大鼠的闭合臂持续时间与控制大鼠相比增加, 这意味着模型大鼠在闭合臂上花费的时间更多。注: p < 0.05 (*);p <0.01 (**);p < 0.001 (**)。<a href="/files/ftp_upload/57362/57362fig4large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/57362/57362fig5.jpg"> 图 5: 详尽的游泳试验分析.数据显示为中 IQR (n = 10), 并报告为p < 0.05 (*), p < 0.01 (**), p < 0.001 (**)。意义是由曼-惠特尼 U 测试, z =-3.326, p = 0.001 确定。模型大鼠的游泳时间 (n = 10) 比对照鼠明显短 (n = 10)。
<img alt="Figure 6" class="xfigimg" src="/files/ftp_upload/57362/57362fig6.jpg"> 图 6: 中央神经递质含量分析.(A) 对 DA 内容的比较。数据显示为均值电子扫描电镜 (n = 10)。具有等方差 (F = 0.088, p = 0.771 > 0.05), 意义由 t 检验, t = 3.717, p = 0.002 < 0.01 决定。模型大鼠 (n = 10) 与对照鼠 (n = 10) 相比, 两 hypothalamuses 的 DA 含量均呈下降趋势。(B) 5 HT 含量的比较。数据显示为均值电子扫描电镜 (n = 10)。以不等方差 (F = 5.282, p = 0.034 < 0.05), 重要性由 t 测试, t =-2.997, p = 0.012 < 0.05 决定。在模型大鼠中, hypothalamuses 的 5 HT 含量与对照大鼠相比均有下降趋势。比率比较。数据显示为中 IQR (n = 10)。意义是由曼-惠特尼 U 测试, z =-3.175, p = 0.001 确定。模型大鼠与对照大鼠相比, DA-5-HT 的比值显著降低。注: p < 0.05 (*);p <0.01 (**);p < 0.001 (**)。

Watch this Scientific Journal Video about A Rat Model of Central Fatigue Using a Modified Multiple Platform Method at JoVE.com

A Rat Model of Central Fatigue Using a Modified Multiple Platform Method

在这里, 我们提出了一个使用改进的多平台方法 (MMPM) 来引入大鼠中心疲劳模型的协议。

基于改进多平台法的大鼠中心疲劳模型

In this article, we introduced a rat model of central fatigue using the modified multiple platform method (MMPM). The Multiple Platform box was designed ...

a-rat-model-central-fatigue-using-modified-multiple-platform

Nanyang Technological University

Research

JoVE Journal

Neuroscience

8.9K 次观看.  Beijing University of Chinese Medicine. 在这里, 我们提出了一个使用改进的多平台方法 (MMPM) 来引入大鼠中心疲劳模型的协议。

视频: A Rat Model of Central Fatigue Using a Modified Multiple Platform Method

A Mouse Model of Fatigue Induced by Peripheral Irradiation

Cancer-related fatigue (CRF) is a distressing and costly condition that often affects patients receiving cancer treatments, including radiation therapy. Here we describe a method using targeted peripheral irradiation to induce fatigue-like behavior in mice. With appropriate shielding, the irradiation targets the lower abdominal/pelvic region of the mouse, sparing the brain, in an effort to model radiation treatment received by individuals with pelvic cancers. We deliver an irradiation dose that is sufficient to induce fatigue-like behavior in mice, measured by voluntary wheel-running activity (VWRA), without causing obvious morbidity. Since wheel running is a normal, voluntary behavior in mice, its use should have little confounding effect on other behavioral tests or biological measures. Hence, wheel running can be used as a feasible outcome measure in understanding the behavioral and biological correlates of fatigue. CRF is a complex condition with frequent comorbidities, and likely has causes related both to cancer and its various treatments. The methods described in this paper are useful for investigating radiation-induced changes that contribute to the development of CRF and, more generally, to explore the biological networks that can explain the development and persistence of a peripherally-triggered but centrally-driven behavior like fatigue.

We describe a method using targeted peripheral irradiation to induce fatigue-like behavior in mice. The selected non-lethal irradiation dose leads to a week-long reduction in voluntary wheel-running activity.

疲劳的小鼠模型的外围照射引起

Cancer-related fatigue (CRF) is a distressing and costly condition that often affects patients receiving cancer treatments, including radiation therapy. ...

科研

癌症研究

Force and Position Control in Humans - The Role of Augmented Feedback

During motor behaviour, humans interact with the environment by for example manipulating objects and this is only possible because sensory feedback is constantly integrated into the central nervous system and these sensory inputs need to be weighted in order meet the task specific goals. Additional feedback presented as augmented feedback was shown to have an impact on motor control and motor learning. A number of studies investigated whether force or position feedback has an influence on motor control and neural activation. However, as in the previous studies the presentation of the force and position feedback was always identical, a recent study assessed whether not only the content but also the interpretation of the feedback has an influence on the time to fatigue of a sustained submaximal contraction and the (inhibitory) activity of the primary motor cortex using subthreshold transcranial magnetic stimulation. This paper describes one possible way to investigate the influence of the interpretation of feedback on motor behaviour by investigating the time to fatigue of submaximal sustained contractions together with the neuromuscular adaptations that can be investigated using surface EMG. Furthermore, the current protocol also describes how motor cortical (inhibitory) activity can be investigated using subthreshold TMS, a method known to act solely on the cortical level. The results show that when participants interpret the feedback as position feedback, they display a significantly shorter time to fatigue of a submaximal sustained contraction. Furthermore, subjects also displayed an increased inhibitory activity of the primary cortex when they believed to receive position feedback compared when they believed to receive force feedback. Accordingly, the results show that interpretation of feedback results in differences on a behavioural level (time to fatigue) that is also reflected in interpretation-specific differences in the amount of inhibitory M1 activity.

Controlling an identical movement with position or force feedback results in different neural activation and motor behavior. This protocol describes how to investigate behavioral changes by looking at neuromuscular fatigue and how to evaluate motor cortical (inhibitory) activity using subthreshold TMS with respect to the interpretation of augmented feedback.

力和位置控制人类 - 增强反馈的作用

During motor behaviour, humans interact with the environment by for example manipulating objects and this is only possible because sensory feedback is ...

行为学

The Treadmill Fatigue Test: A Simple, High-throughput Assay of Fatigue-like Behavior for the Mouse

Fatigue is a prominent symptom in many diseases and disorders and reduces quality of life for many people. The lack of clear pathogenesis and failure of current interventions to adequately treat fatigue in all patients leaves a need for new treatment options. Despite the therapeutic need and importance of preclinical research in helping identify promising novel treatments, few preclinical assays of fatigue are available. Moreover, the most common preclinical assay used to assess fatigue-like behavior, voluntary wheel running, is not suitable for use with some strains of mice, may not be sensitive to drugs that reduce fatigue, and has relatively low throughput. The current protocol describes a novel, non-voluntary preclinical assay of fatigue-like behavior, the treadmill fatigue test, and provides evidence of its efficacy in detecting fatigue-like behavior in mice treated with a chemotherapy drug known to cause fatigue in humans and fatigue-like behavior in animals. This assay may be a beneficial alternative to wheel running, as fatigue-like behavior and potential interventions can be assessed in a greater number of mice over a shorter time frame, thus permitting faster discovery of new therapeutic options.

Fatigue is a common, undertreated and frequently poorly-understood symptom in many diseases and disorders. New preclinical assays of fatigue may help to improve current understanding and future treatment of fatigue. To that end, the current protocol provides a novel means of measuring fatigue-like behavior in the mouse.

跑步机疲劳试验:疲劳类似的行为，为鼠标操作简单，高通量检测

Fatigue is a prominent symptom in many diseases and disorders and reduces quality of life for many people. The lack of clear pathogenesis and failure of ...

Sit-to-stand-and-walk from 120% Knee Height: A Novel Approach to Assess Dynamic Postural Control Independent of Lead-limb

Individuals with sensorimotor pathology e.g., stroke have difficulty executing the common task of rising from sitting and initiating gait (sit-to-walk: STW). Thus, in clinical rehabilitation separation of sit-to-stand and gait initiation - termed sit-to-stand-and-walk (STSW) - is usual. However, a standardized STSW protocol with a clearly defined analytical approach suitable for pathological assessment has yet to be defined.
Hence, a goal-orientated protocol is defined that is suitable for healthy and compromised individuals by requiring the rising phase to be initiated from 120% knee height with a wide base of support independent of lead limb. Optical capture of three-dimensional (3D) segmental movement trajectories, and force platforms to yield two-dimensional (2D) center-of-pressure (COP) trajectories permit tracking of the horizontal distance between COP and whole-body-center-of-mass (BCOM), the decrease of which increases positional stability but is proposed to represent poor dynamic postural control.
BCOM-COP distance is expressed with and without normalization to subjects&#39; leg length. Whilst COP-BCOM distances vary through STSW, normalized data at the key movement events of seat-off and initial toe-off (TO1) during steps 1 and 2 have low intra and inter subject variability in 5 repeated trials performed by 10 young healthy individuals. Thus, comparing COP-BCOM distance at key events during performance of an STSW paradigm between patients with upper motor neuron injury, or other compromised patient groups, and normative data in young healthy individuals is a novel methodology for evaluation of dynamic postural stability.

Here, we present a novel protocol to measure positional stability at key events during the sit-to-stand-to-walk using the center-of-pressure to the whole-body-center-of-mass distance. This was derived from the force platform and three-dimensional motion-capture technology. The paradigm is reliable and can be utilized for the assessment of neurologically compromised individuals.

坐姿到站姿和步行距离120％膝盖高度:新方法来评估铅肢体的动态姿势控制独立

Individuals with sensorimotor pathology e.g., stroke have difficulty executing the common task of rising from sitting and initiating gait (sit-to-walk: ...

Measuring the Motor Aspect of Cancer-Related Fatigue using a Handheld Dynamometer

Cancer-related fatigue (CRF) is commonly reported by patients both during and after receiving treatment for cancer. Current CRF diagnoses rely on self-report questionnaires which are subject to report and recall biases. Objective measurements using a handheld dynamometer, or handgrip device, have been shown in recent studies to correlate significantly with subjective self-reported fatigue scores. However, variations of both the handgrip fatigue test and fatigue index calculations exist in the literature. The lack of standardized methods limits the utilization of the handgrip fatigue test in the clinical and research settings. In this study, we provide detailed methods for administering the physical fatigue test and calculating the fatigue index. These methods should supplement existing self-reported fatigue questionnaires and help clinicians assess fatigue symptom severity in an objective and quantitative manner.

Simple and accessible methods were developed to measure the motor aspect of cancer-related fatigue objectively and quantitatively. We describe, in detail, ways to administer the physical fatigue test using a simple handgrip device as well as methods to calculate fatigue indices.

使用手持测功机测量癌症相关疲劳的马达方面

Cancer-related fatigue (CRF) is commonly reported by patients both during and after receiving treatment for cancer. Current CRF diagnoses rely on ...

Revised and Neuroimaging-Compatible Versions of the Dual Task Screen

Dual task paradigms simultaneously assess motor and cognitive abilities, and they can detect subtle, residual impairments in athletes with recent mild traumatic brain injury (mTBI). However, past dual task paradigms have focused solely on lower extremity skills and have relied on cumbersome, expensive laboratory equipment - thus limiting their practicality for everyday mTBI evaluation. Subsequently, we developed the Dual Task Screen (DTS), which takes &#60;10 minutes to administer and score, uses low-cost portable equipment, and includes lower extremity (LE) and upper extremity (UE) subtasks. The purpose of this manuscript was twofold. First, we describe the administration protocol for the revised DTS, which we revised to address the limitations of the original DTS. Specifically, the revisions included additions of smart devices to acquire more detailed gait data and inclusion of single cognitive conditions to test for disrupted cognitive performance under dual task conditions. Importantly, the revised DTS is a measure intended for future clinical use, and we present representative results from three male athletes to illustrate the type of clinical data that can be acquired from the measure. Importantly, we have yet to evaluate the sensitivity and specificity of the revised DTS in athletes with mTBI, which is the next research initiative. The second purpose of this manuscript is to describe a neuroimaging-compatible version of the DTS. We developed this version so we could evaluate the neural underpinnings of single and dual task performance, for a better empirical understanding of the behavioral deficits associated with mTBI. Thus, this manuscript also describes the steps we took to enable simultaneous functional near-infrared spectroscopy (fNIRS) measurement during the DTS, along with how we acquired and completed first-level processing of the fNIRS data.

We developed the original Dual Task Screen (DTS) as a portable, low-cost measure that can evaluate athletes with sports-induced mild traumatic brain injury. We revised the original DTS for future clinical use and developed a neuroimaging-compatible version of the DTS to measure neural underpinnings of single and dual task performance.

双任务屏幕的修订和神经成像兼容版本

Dual task paradigms simultaneously assess motor and cognitive abilities, and they can detect subtle, residual impairments in athletes with recent mild ...

集成 VR 和生物传感器进行定量运动肌肉活动评估

Setup for the Quantitative Assessment of Motion and Muscle Activity During a Virtual Modified Box and Block Test

The ability to move allows us to interact with the world. When this ability is impaired, it can significantly reduce one's quality of life and independence and may lead to complications. The importance of remote patient evaluation and rehabilitation has recently grown due to limited access to in-person services. For example, the COVID-19 pandemic unexpectedly resulted in strict regulations, reducing access to non-emergent healthcare services. Additionally, remote care offers an opportunity to address healthcare disparities in rural, underserved, and low-income areas where access to services remains limited. 
Improving accessibility through remote care options would limit the number of hospital or specialist visits and render routine care more affordable. Finally, the use of readily available commercial consumer electronics for at-home care can enhance patient outcomes due to improved quantitative observation of symptoms, treatment efficacy, and therapy dosage. While remote care is a promising means to address these issues, there is a crucial need to quantitatively characterize motor impairment for such applications. The following protocol seeks to address this knowledge gap to enable clinicians and researchers to obtain high-resolution data on complex movement and underlying muscle activity. The ultimate goal is to develop a protocol for remote administration of functional clinical tests. 
Here, participants were instructed to perform a medically-inspired Box and Block task (BBT), which is frequently used to assess hand function. This task requires subjects to transport standardized cubes between two compartments separated by a barrier. We implemented a modified BBT in virtual reality to demonstrate the potential of developing remote assessment protocols. Muscle activation was captured for each subject using surface electromyography. This protocol allowed for the acquisition of high-quality data to better characterize movement impairment in a detailed and quantitative manner. Ultimately, these data have the potential to be used to develop protocols for virtual rehabilitation and remote patient monitoring.

作者聚焦:利用虚拟现实和肌电图增强远程康复

The protocol described here aims to enhance the quantitative evaluation of upper limb deficits, with the goal of developing additional technology for remote assessment both in the clinic and at home. Virtual reality and biosensor technologies are combined with standard clinical techniques to provide insights into the functioning of the neuromuscular system.

用于在虚拟改良的 Box and Block 测试期间定量评估运动和肌肉活动的设置

The ability to move allows us to interact with the world. When this ability is impaired, it can significantly reduce one's quality of life and ...

工程学

Determining The Electromyographic Fatigue Threshold Following a Single Visit Exercise Test

Theoretically, the electromyographic (EMG) fatigue threshold is the exercise intensity an individual can maintain indefinitely without the need to recruit more motor units which is associated with an increase in the EMG amplitude. Although different protocols have been used to estimate the EMG fatigue threshold they require multiple visits which are impractical for a clinical setting. Here, we present a protocol for estimating the EMG fatigue threshold for cycle ergometry which requires a single visit. This protocol is simple, convenient, and completed within 15-20 min, therefore, has the potential to be translated into a tool that clinicians can use in exercise prescription.

This protocol describes the electromyographic fatigue threshold which demarcates between nonfatiguing and fatiguing exercise workloads. This information could be used to develop a more individualized training program.

确定肌电疲劳阈值以下单次访问运动试验

Theoretically, the electromyographic (EMG) fatigue threshold is the exercise intensity an individual can maintain indefinitely without the need to recruit ...

基于改进多平台法的大鼠中心疲劳模型

摘要

探索更多视频

此视频中的章节

疲劳的小鼠模型的外围照射引起

力和位置控制人类 - 增强反馈的作用

跑步机疲劳试验:疲劳类似的行为，为鼠标操作简单，高通量检测

坐姿到站姿和步行距离120％膝盖高度:新方法来评估铅肢体的动态姿势控制独立

使用手持测功机测量癌症相关疲劳的马达方面

双任务屏幕的修订和神经成像兼容版本

用于在虚拟改良的 Box and Block 测试期间定量评估运动和肌肉活动的设置

用于在虚拟改良的 Box and Block 测试期间定量评估运动和肌肉活动的设置

用于在虚拟改良的 Box and Block 测试期间定量评估运动和肌肉活动的设置

确定肌电疲劳阈值以下单次访问运动试验