这项工作得到了河北省科技计划项目（NO.E2020100001和NO.22372502D）和石家庄市高水平科技创新创业人才项目（NO.07202203）的支持。

动物实验经河北夷陵医学研究院动物研究伦理委员会批准（批准文号：N2022148）。将体重18-22 g的雄性C57BL/6J小鼠（见 材料表）安置在河北夷陵医学研究所新药评价中心。他们每天获得正常的食物和干净的水，并暴露在人造光下12小时。房间的温度控制在20-26°C，相对湿度为40%-70%。
1.慢性缺氧小鼠模型的建立（图1）
<ol><li>在开始实验之前，准备正常大气压下的动物笼子和连续低氧环境的笼子。利用自动气体控制输送系统用纯氧和氮气的混合物冲洗腔室，从而建立连续的低氧环境。 注意： 该系统被编程为控制电磁阀开关，从而确保在时间和浓度方面精确输送气体。</li>
	<li>将小鼠随机分为三组：对照组（Con），模型组（CH）和电针灸组（EA + CH）。将对照和模型/电针灸小鼠分别放置在两个笼子中，每个笼子有10只小鼠。将光照周期保持在 12 小时/12 小时（亮/暗）。 注意：对照组（Con）未诱导治疗或缺氧。模型组（CH）由慢性缺氧小鼠组成。电针灸组（EA + CH）包括用电针治疗的缺氧诱导小鼠。</li>
	<li>对于慢性缺氧，利用数字氧气计调节气体流速并保持 10% 的氧气浓度，建立低氧室的参数。在上午9：00将动物置于低氧室中，并在下午5：00将其移出，从而每天连续暴露于低氧8小时，持续3个月。 注意： 在设置氮气输送以降低氧气浓度时，建议缓慢进行以防止立即过量引入氮气，因为这会导致动物死亡。</li>
	<li>使用组织学检查和行为测试评估慢性缺氧诱导的认知功能障碍模型：开放场地测试15 和水迷宫测试16。</li>
</ol>2.麻醉（图2）
<ol><li>准备小动物麻醉机（见 材料表）和恒温加热垫。 注意：在麻醉期间，动物容易出现体温过低，这强调了使用恒温加热垫进行绝缘的必要性。</li>
	<li>将小鼠置于麻醉诱导箱中，并用2%-2.5%异氟醚在氧气中快速诱导约1分钟。 注意：这种短期预处理是确保小鼠能够在低浓度剂量下长时间茁壮成长的关键步骤。</li>
	<li>一旦它们的兴奋性降低，捏住鼠标的脚趾以检查其反射。然后，将鼠标转移到恒温加热垫（37°C）上。</li>
	<li>将麻醉流速调节至约0.5%浓度。将麻醉机连接到鼠标的嘴和鼻子。在确保维持麻醉的同时进行电针治疗。 注意：当小鼠停止眨眼时，麻醉效果得到确认。麻醉效果可以持续至少30分钟。</li>
</ol>3.电针治疗
<ol><li>为有效改善认知功能障碍，应根据中医理论和临床经验，选择特定的穴位，如百会（GV20）、神庭（GV24）、双侧祖三里（ST36）（图3）。在建模过程完成前 2 周进行电针治疗。<ol><li>将 GV20 穴位定位在前额的中线上，在连接耳尖的线的中点7.针灸针插入的深度必须为 2 毫米。</li>
		<li>将 GV24 穴位定位在前额中线鼠标眼睛中点正上方 1.3 毫米处17.针灸针插入的深度必须为 2 毫米。</li>
		<li>将 ST36 穴位定位在膝关节外侧，腓骨头下方约 2 毫米处18,19。针灸针插入的深度必须为3-4毫米。</li>
	</ol></li>
	<li>为该过程准备一次性针灸针（见 材料表）和电针灸设备（见 材料表）（图4）。</li>
	<li>将小鼠置于俯卧位，在0.5%异氟烷的轻度麻醉下，确保其头部和四肢固定。用右手握住一根不锈钢针（直径：0.18 毫米;长度：7 毫米），用拇指、食指和中指。</li>
	<li>在 GV20 和 GV24 穴位横向进行针灸 2 mm 深度，用左手抬起小鼠头部的皮肤。通过触摸鼠标膝关节外侧的腓骨头并用左手拇指按压皮肤，垂直刺穿 ST36 穴位 3-4 毫米深。 注意： 对于位于头部的穴位，建议按 GV24 和 GV20 的顺序插入针头。该订单有助于操作便利。穴位是离散的解剖位置，而不是静止的穴位。因此，针头插入角度的轻微偏差对治疗效果没有影响，在临床环境中接受电针治疗的患者中也类似。</li>
	<li>将电子针灸装置连接到针头上，GV20和左侧ST36连接到一个电极组，GV24和右侧ST36连接到另一个电极组（图4）。选择连续波模式，电流强度为 2 mA，频率为 2 Hz20,21。通过观察穴位的局部轻度震颤和鼠标的安静耐受性来确认理想的治疗方法。<ol><li>连接电针灸器械时，连接针的近端。这有助于最大限度地减少连接线重量造成的影响，从而防止针头脱落。如果需要，使用胶带固定水平插入的针头和连接线。</li>
	</ol></li>
	<li>每天进行每日治疗，每次30分钟，连续6天，每个治疗周期之间休息一天。</li>
</ol>4. 开放场测试（图5）
注：开放场测试是一种常规方法，用于评估实验动物在新奇和不熟悉环境中的自主行为、探索行为、认知能力和焦虑行为22.它由一个露天反应箱和一个记录装置组成。
<ol><li>要进行测试，请准备一个尺寸为 50 厘米× 50 厘米× 30 厘米的白壁立方体，底部分为 25 个相等的正方形，尺寸为 10 厘米× 10 厘米。</li>
	<li>将鼠标放入空旷的野外反应箱中进行驯化。让鼠标在适应期探索测试室并熟悉新环境。在使小鼠适应实验环境1小时后进行露天试验。 注意：这保证了由环境变化引起的焦虑或压力的最大化，从而在随后的行为评估中实现更精确的结果。</li>
	<li>将鼠标放在盒子的中央，让鼠标适应环境2分钟后监测10分钟。<ol><li>使用视频跟踪系统（见 材料表）记录鼠标的移动轨迹、行进的总距离、在中心区域花费的时间、穿过中心区域的速度以及测试期间进入中心区域的次数。</li>
		<li>按照视频跟踪系统产品手册中的说明进行相关操作。每只鼠标都经过一次测试，并从盒子内的同一位置开始探索。</li>
		<li>每次测试后，用 75% 乙醇清洁开放的场地箱，以防止使用鼠标时因气味干扰而导致的任何错误结果。</li>
	</ol></li>
</ol>5. 水迷宫（图5）
注：在涉及小鼠的实验中，水迷宫测试经常被用作行为评估工具，以评估其空间学习和记忆能力23。
<ol><li>准备一个直径为120厘米，深度为30厘米的圆形水箱。将水箱分为四个相等的象限：I、II、III 和 IV。如果在实验中使用黑鼠，请使用白色水箱;对于白鼠，请使用黑色水箱。</li>
	<li>在圆形水箱周围放置窗帘，以防止鼠标在测试过程中看到研究人员。</li>
	<li>在水箱的顶面放置不同的标记，作为空间定位的视觉提示。确保这些标记在整个实验过程中保持静止，以保持一致性。</li>
	<li>在水箱的象限III中放置一个直径为10厘米的圆形平台作为指定的目标区域。确保平台可以轻松移动并固定在任何所需位置。</li>
	<li>在整个实验过程中，将水引入水箱，同时保持22-24°C的温度范围。<ol><li>确保水位始终保持在目标平台上方 1 厘米。在水中加入 20% 浓度的无毒二氧化钛，以在黑色小鼠和白色背景之间实现鲜明对比。这种对比度有助于相机记录鼠标的移动和相关参数。</li>
	</ol></li>
	<li>通过依次将每只鼠标放置在象限 I、II、III 和 IV 中，进行为期 5 天的连续空间探索测试。<ol><li>将鼠标朝向墙壁。远离迷宫，以防止鼠标使用实验者的位置作为参考点。记录鼠标找到平台所需的时间。</li>
		<li>如果鼠标未能在 90 秒内找到水下平台，请将鼠标引导到平台并提供 30 秒的学习时间。此外，将延迟期记录为 90 秒。</li>
		<li>如果鼠标在 90 秒内找到水下平台，让它在平台上停留 10 秒进行学习，然后再将其从水箱中取出。</li>
		<li>用毛巾擦干鼠标并将其放回笼子。</li>
		<li>每 20 分钟轮换每个鼠标在每个象限中的位置。使用视频跟踪系统（见 材料表）记录每只鼠标的游泳距离、速度和找到平台所需的时间（延迟期），按照产品手册中的说明执行相关操作。</li>
		<li>在第 1 天将平台设置在水面以上 1 厘米处。在第 2-5 天将平台放置在水面以下 1 厘米的深度。</li>
	</ol></li>
	<li>在第 6 天，将平台从目标象限中移除并进行空间探索测试。<ol><li>将鼠标放在象限 I 中自由探索 90 秒。计算机记录鼠标的游泳轨迹、在目标象限停留的时间以及它穿过平台的次数。 注意：为了尽量减少人为因素引起的实验误差，在水迷宫实验中保持参考点的位置固定是很重要的。此外，实验者在将小鼠放入水中后应立即撤退。实验完成后，应用毛巾擦干小鼠并放回笼子中以保持温暖。</li>
	</ol></li>
</ol>6.苏木精和伊红（HE）染色 （图6）
注意：海马区域的组织学检查有助于评估缺氧模型的建立并确定针灸治疗的疗效。
<ol><li>行为实验后，用腹膜内注射20mg / kg戊巴比妥钠麻醉小鼠，并用10%多聚甲醛溶液灌注（见 材料表）以确保完全灌注。分离脑组织，在室温（RT）下浸入10%多聚甲醛溶液中3天，达到固定效果。</li>
	<li>将大脑样本放入嵌入盒中。随后，用流水洗涤处理过的脑样品6小时。</li>
	<li>使用自动组织处理器使用一系列浓度递增的酒精溶液对样品进行脱水，即 60% 乙醇 1 小时、70% 乙醇 1 小时、90% 乙醇 1 小时、95% 乙醇 2 小时，最后 100% 乙醇 2 小时。</li>
	<li>将组织标本浸入二甲苯中2小时以达到透明度。随后，在脱水过程完成后，将透化样品转移到加热至60°C的石蜡中3小时。最后，将它们嵌入自动处理器中。</li>
	<li>使用旋转切片机获得 4 μm 切片。随后，对切片进行苏木精染色3-8分钟，然后进行曙红染色1-3分钟。</li>
	<li>依次将染色切片转移到纯酒精和二甲苯的分离容器中。然后，用中性胶密封并固定染色切片，准备在光学显微镜下进行病理检查。</li>
	<li>使用载玻片扫描仪（参见 材料表）扫描切片。随后，利用观察软件获得海马区域的HE染色结果。比较神经元的排列和神经元核的凝聚。</li>
</ol>

针灸慢性缺氧小鼠的认知改善

作者没有什么可透露的。

针灸是一种起源于2000多年前中国的非药物医学实践，涉及将细针插入身体的特定穴位，称为穴位。这些点被认为通过通道或经络连接，身体的生命能量或“气”通过这些通道或经络流动24.通过刺激这些穴位，针灸旨在恢复身体的平衡与和谐。它已被证明可以有效治疗各种疾病，包括慢性疼痛、焦虑/抑郁、消化问题、月经来潮和呼吸系统疾病 25,26,27,28,29。</s...

全球人口目前正面临严重的老龄化问题，导致认知障碍的患病率迅速增加。全球认知障碍的发病率约为每 1000 人年 53.97 例1。血管功能障碍或循环/呼吸系统疾病引起的慢性脑缺氧仍然是年龄相关性痴呆的主要危险因素之一2。先前的研究表明，脑缺氧可以通过改变 BACE1 表达来增加淀粉样蛋白β沉积3。此外，缺氧与神经胶质细胞失调和神经炎症有关 4,5。尽管这个问题越来越严重，但目前缺乏有效的西药来预防慢性缺氧引起的认知能力下降。非药物中医，特别是针灸，已被用于治疗认知障碍数千年，并在缓解神经退行性疾病方面显示出可喜的效果6,7。百慧穴、神庭穴、祖三里穴位是治疗认知功能障碍的有效穴位8,9。临床研究表明，电针疗法可显著改善血管性认知障碍患者的蒙特利尔认知评估（MoCA）和简易精神状态检查（MMSE）评分，有效改善认知功能障碍8。尽管研究表明针灸可以显着增强动脉结扎大鼠的记忆能力——急性脑缺氧模型10，急性脑缺氧模型，但没有关于针灸对任何具有慢性缺氧诱导认知障碍的啮齿动物模型的影响的报道。缺乏对该机制的研究极大地阻碍了其临床应用。
先前的研究表明，将大鼠置于缺氧环境中 8 周可以显着提高大脑中的氧化应激和炎症水平，导致记忆功能下降11.本研究旨在探讨针灸对啮齿动物模型的影响，以加深我们的理解。然而，值得注意的是，在啮齿动物的针灸治疗期间通常需要麻醉，因为在反复刺激期间可能会引起躁动。长时间麻醉会显着影响小鼠的认知功能，因为大多数麻醉药物会抑制神经活动并阻碍信息处理，导致行为缺陷12。几项研究表明，施用 2.5% 七氟醚持续 6 小时会显着损害小鼠的空间记忆、学习能力和注意力13。此外，有证据表明，高剂量麻醉可能导致小鼠神经元死亡或神经损伤14。因此，必须确定一种合适的方法，以尽量减少麻醉的总使用量。在这项研究中，我们介绍了一种有效的针灸方法来治疗认知障碍小鼠，以及评估其记忆能力的行为测试。重要的是，我们提出了一种改进的预处理麻醉技术，可以有效减少实验期间施用的麻醉总剂量。

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			<td>10% paraformaldehyde solution</td>
			<td>Bioroyee (Beijing) Biotechnology Co., Ltd</td>
			<td>RL3234</td>
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			<td>C75BL/6J mice</td>
			<td>BEIJING HFK BIOSCIENCE CO.,LTD</td>
			<td>No.110322220103041767</td>
			<td>Gender: Male,&#160; Weight: 18&#8211;22 g</td>
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			<td>Electroacupuncture device</td>
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			<td>KWD-808 I</td>
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			<td>Suzhou Medical Appliance Factory</td>
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			<td>Isoflurane</td>
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			<td>Small animal anesthesia machine</td>
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			<td>YL-LE-A106</td>
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acupuncture treatment in a mouse model of chronic hypoxia-induced cognitive dysfunction

中枢神经紊乱的治疗一直对医学领域构成重大挑战。针灸是一种植根于传统中医的非药物实践，需要将细针插入身体的精确穴位，通常用于治疗各种疾病。最近，针灸已成为治疗一系列神经系统疾病（包括焦虑症和呼吸系统疾病）的有前途的治疗干预措施。然而，针灸治疗慢性缺氧引起的认知功能障碍的潜力尚未得到探索。本文提出了一个全面的方案，用于建立慢性缺氧诱导的认知障碍小鼠模型、进行轻度麻醉、进行针灸治疗以及使用开放场测试和水迷宫评估行为变化和记忆能力。分步协议提供了有关准确定位和定位穴位和针头以改善认知的详细说明。通过采用该协议，研究人员可以进行系统研究，以彻底评估针灸治疗认知功能障碍的潜力。

在开阔场实验中表征小鼠运动轨迹 轨迹图显示，正常组中的小鼠表现出在陌生环境中探索的强烈倾向。他们的活动轨迹主要集中在角落，同时覆盖整个空地（左图）。相比之下，长期缺氧模型组的小鼠表现出探索新环境的欲望显着减弱。它们主要在角落里徘徊，没有表现出任何朝向空旷场地中心（中间面板）的探索行为。针灸治疗后，缺氧诱导的小鼠的探索活动得到改善，并且它?...

Watch this Scientific Journal Video about Acupuncture Treatment in a Mouse Model of Chronic Hypoxia-Induced Cognitive Dysfunction at JoVE.com

作者聚焦：改进增强小鼠针刺研究的麻醉方案

在这里，我们描述了一种在慢性缺氧小鼠模型上实施轻度麻醉和针灸治疗的方案，并进行行为测试以评估治疗后的认知改变。

针灸治疗慢性缺氧诱导认知功能障碍小鼠模型

The treatment of central nervous disorders has consistently posed significant challenges to the medical field. Acupuncture, a non-pharmacological practice ...

选择正确的麻醉方案对于进行小鼠针灸至关重要。由于与长时间麻醉相关的潜在认知障碍，它代表了一个重要的初始挑战，需要仔细考虑，以尽量减少手术过程中小鼠的认知功能风险。在这项研究中，我们提出了一种改进的方案，有助于最大限度地减少因过度麻醉剂量引起的功能和行为异常，并提高实验的准确性。未来，我们的实验室旨在研究针灸的潜在机制，尽管它对行为和病理学有可观察到的治疗效果。我们的研究将集中在解开针灸上，以更深入地了解其机制。

acupuncture-treatment-mouse-model-chronic-hypoxia-induced-cognitive

Research

JoVE Journal

Behavior

Acupuncture Treatment in a Mouse Model of Chronic Hypoxia-Induced Cognitive Dysfunction

807 次观看.  Heilongjiang University of Chinese Medicine. 选择正确的麻醉方案对于进行小鼠针灸至关重要。由于与长时间麻醉相关的潜在认知障碍，它代表了一个重要的初始挑战，需要仔细考虑，以尽量减少手术过程中小鼠的认知功能风险。在这项研究中，我们提出了一种改进的方案，有助于最大限度地减少因过度麻醉剂量引起的功能和行为异常，并提高实验的准确性。未来，我们的实验室旨在研究针灸的潜在机制，尽管它?...

视频: 作者聚焦：改进增强小鼠针刺研究的麻醉方案

Using Chronic Social Stress to Model Postpartum Depression in Lactating Rodents

Exposure to chronic stress is a reliable predictor of depressive disorders, and social stress is a common ethologically relevant stressor in both animals and humans. However, many animal models of depression were developed in males and are not applicable or effective in studies of postpartum females. Recent studies have reported significant effects of chronic social stress during lactation, an ethologically relevant and effective stressor, on maternal behavior, growth, and behavioral neuroendocrinology. This manuscript will describe this chronic social stress paradigm using repeated exposure of a lactating dam to a novel male intruder, and the assessment of the behavioral, physiological, and neuroendocrine effects of this model. Chronic social stress (CSS) is a valuable model for studying the effects of stress on the behavior and physiology of the dam as well as her offspring and future generations. The exposure of pups to CSS can also be used as an early life stress that has long term effects on behavior, physiology, and neuroendocrinology.

This article describes the use of chronic resident intruder social stress as an ethologically relevant paradigm to model postpartum depression and anxiety in lactating rodents.

哺乳期鼠害使用慢性社会压力型号产后抑郁症

Exposure to chronic stress is a reliable predictor of depressive disorders, and social stress is a common ethologically relevant stressor in both animals ...

科研

行为学

The Crossmodal Congruency Task as a Means to Obtain an Objective Behavioral Measure in the Rubber Hand Illusion Paradigm

The rubber hand illusion (RHI) is a popular experimental paradigm. Participants view touch on an artificial rubber hand while the participants' own hidden hand is touched. If the viewed and felt touches are given at the same time then this is sufficient to induce the compelling experience that the rubber hand is one's own hand. The RHI can be used to investigate exactly how the brain constructs distinct body representations for one's own body. Such representations are crucial for successful interactions with the external world. To obtain a subjective measure of the RHI, researchers typically ask participants to rate statements such as "I felt as if the rubber hand were my hand". Here we demonstrate how the crossmodal congruency task can be used to obtain an objective behavioral measure within this paradigm.
The variant of the crossmodal congruency task we employ involves the presentation of tactile targets and visual distractors. Targets and distractors are spatially congruent (i.e. same finger) on some trials and incongruent (i.e. different finger) on others. The difference in performance between incongruent and congruent trials - the crossmodal congruency effect (CCE) - indexes multisensory interactions. Importantly, the CCE is modulated both by viewing a hand as well as the synchrony of viewed and felt touch which are both crucial factors for the RHI.
The use of the crossmodal congruency task within the RHI paradigm has several advantages. It is a simple behavioral measure which can be repeated many times and which can be obtained during the illusion while participants view the artificial hand. Furthermore, this measure is not susceptible to observer and experimenter biases. The combination of the RHI paradigm with the crossmodal congruency task allows in particular for the investigation of multisensory processes which are critical for modulations of body representations as in the RHI.

We demonstrate how an objective measure can be employed in the widely employed rubber hand illusion paradigm. This measure is obtained by modifying the well-established crossmodal congruency task. This task allows the investigation of multisensory processes which are critical for modulations of body representations as in the rubber hand illusion.

一致性的任务作为一种手段来获取目标行为的措施中的橡胶手错觉范式Crossmodal

The rubber hand illusion (RHI) is a popular experimental paradigm. Participants view touch on an artificial rubber hand while the participants' own hidden ...

A Protocol of Manual Tests to Measure Sensation and Pain in Humans

Numerous qualitative and quantitative techniques can be used to test sensory nerves and pain in both research and clinical settings. The current study demonstrates a quantitative sensory testing protocol using techniques to measure tactile sensation and pain threshold for pressure and heat using portable and easily accessed equipment. These techniques and equipment are ideal for new laboratories and clinics where cost is a concern or a limiting factor. We demonstrate measurement techniques for the following: cutaneous mechanical sensitivity on the arms and legs (von-Frey filaments), radiant and contact heat sensitivity (with both threshold and qualitative assessments using the Visual Analog Scale (VAS)), and mechanical pressure sensitivity (algometer, with both threshold and the VAS). The techniques and equipment described and demonstrated here can be easily purchased, stored, and transported by most clinics and research laboratories around the world. A limitation of this approach is a lack of automation or computer control. Thus, these processes can be more labor intensive in terms of personnel training and data recording than the more sophisticated equipment. We provide a set of reliability data for the demonstrated techniques. From our description, a new laboratory should be able to set up and run these tests and to develop their own internal reliability data.

The goal of this procedure is to demonstrate a battery of quantitative techniques for sensory and pain measurement in humans. The equipment and techniques described are commonly found in pain clinics or are easy to obtain.

手动测试的协议来衡量感觉和疼痛人类

Numerous qualitative and quantitative techniques can be used to test sensory nerves and pain in both research and clinical settings. The current study ...

Visualizing Motion Patterns in Acupuncture Manipulation

Acupuncture manipulation varies widely among practitioners in clinical settings, and it is difficult to teach novice students how to perform acupuncture manipulation techniques skillfully. The Acupuncture Manipulation Education System (AMES) is an open source software system designed to enhance acupuncture manipulation skills using visual feedback. Using a phantom acupoint and motion sensor, our method for acupuncture manipulation training provides visual feedback regarding the actual movement of the student's acupuncture manipulation in addition to the optimal or intended movement, regardless of whether the manipulation skill is lifting, thrusting, or rotating. Our results show that students could enhance their manipulation skills by training using this method. This video shows the process of manufacturing phantom acupoints and discusses several issues that may require the attention of individuals interested in creating phantom acupoints or operating this system.

Here, we present a protocol for using the Acupuncture Manipulation Education System (AMES) in the training of acupuncture manipulation skills using phantom acupoints.

可视化的运动模式在针刺手法

Acupuncture manipulation varies widely among practitioners in clinical settings, and it is difficult to teach novice students how to perform acupuncture ...

Chronic Sleep Deprivation in Mouse Pups by Means of Gentle Handling

Sleep is critical for proper development and neural plasticity. Moreover, abnormal sleep patterns are characteristic of many neurodevelopmental disorders. Studying how chronic sleep restriction during development can affect adult behavior may add to our understanding of the emergence of behavioral symptoms of neurodevelopmental disorders. While there are many methods that can be used to restrict sleep in rodents including forced locomotion, constant disruption, presentation of an aversive stimulus, or electric shock, many of these methods are very stressful and cannot be used in neonatal mice. Here, we describe gentle handling, a sleep deprivation technique that can be used chronically throughout development and into adulthood to achieve sleep restriction. Gentle handling involves close observation of the mice throughout the sleep deprivation period and requires the researcher to gently prod the animals whenever they are inactive or display behaviors associated with sleep. Coupled with EEG recordings, gentle handling could be used to selectively disrupt a specific phase of sleep such as rapid eye movement (REM) sleep. The technique of gentle handling is a powerful tool for the study of the effects of chronic sleep restriction even in neonatal mice that circumvents many of the more stressful procedures used for sleep deprivation.

We describe a sleep deprivation technique known as gentle handling where investigators gently prod mice any time sleep behavior is observed. This method is a powerful tool that allows researchers to study the effects that chronic sleep restriction throughout development can have on future brain physiology and behavior.

小鼠幼犬慢性睡眠剥夺的温和处理方法

Sleep is critical for proper development and neural plasticity. Moreover, abnormal sleep patterns are characteristic of many neurodevelopmental disorders. ...

Novel Object Recognition and Object Location Behavioral Testing in Mice on a Budget

Ethologically relevant behavioral testing is a critical component of any study that uses mouse models to study the cognitive effects of various physiological or pathological changes. The object location task (OLT) and the novel object recognition task (NORT) are two effective behavioral tasks commonly used to reveal the function and relative health of specific brain regions involved in memory. While both of these tests exploit the inherent preference of mice for the novelty to reveal memory for previously encountered objects, the OLT primarily evaluates spatial learning, which relies heavily on hippocampal activity. The NORT, in contrast, evaluates non-spatial learning of object identity, which relies on multiple brain regions. Both tasks require an open-field-testing arena, objects with equivalent intrinsic value to mice, appropriate environmental cues, and video recording equipment and the software. Commercially available systems, while convenient, can be costly. This manuscript details a simple, cost-effective method for building the arenas and setting up the equipment necessary to perform the OLT and NORT. Furthermore, the manuscript describes an efficient testing protocol that incorporates both OLT and NORT and provides typical methods for data acquisition and analysis, as well as representative results. Successful completion of these tests can provide valuable insight into the memory function of various mouse model systems and appraise the underlying neural regions that support these functions.

Here we provide a protocol which includes comprehensive instructions for the economical establishment of murine object location and novel object recognition behavioral testing, including the design, cost, and construction of required equipment as well as execution of behavioral testing, data collection, and analysis.

一种新的小鼠目标识别与目标定位行为测试

Ethologically relevant behavioral testing is a critical component of any study that uses mouse models to study the cognitive effects of various ...

Analysis of Learning and Memory Ability in an Alzheimer's Disease Mouse Model using the Morris Water Maze

A Morris water maze (MWM) experiment forces experimental animals to swim and learn to find a platform hidden in the water. It is widely used in scientific research to assess the learning and memory of animals. Due to the extensive use of the MWM test, visual experimental protocols are essential for researchers. This manuscript uses the latest studies to introduce the protocol of the MWM test. Alzheimer&#8217; Disease (AD) is characterized by a progressive loss of memory and cognitive function. An alternative and complementary treatment used for AD is Manual Acupuncture (MA). To assess the learning and memory ability of AD model mice, the MWM test was conducted. The visible platform trial, hidden platform trial, probe trial, and reversal trial of MWM were used to evaluate spatial learning and memory ability. In the visible platform trial, the swimming speed and escape latency of mice in different groups was not significantly different. In the hidden platform and reversal trials, the AD group showed a long escape latency. The escape latency decreased significantly after the MA treatment. Low platform crossover number and the proportion of time in the SW quadrant in the probe trial increased after the MA treatment (p &#60; 0.05 or p &#60; 0.01). The results of the MWM tests suggest that MA can effectively improve the spatial learning and memory abilities of AD model mice. Rigorous experimental operations provided assurance of the reliability of the results.

Herein, a protocol to conduct the Morris water maze tests to evaluate the ability of learning and memory of Alzheimer&#8217;s Disease model mice and to assess the effect of manual acupuncture for treating them is described.

使用莫里斯水迷宫对阿尔茨海默氏病小鼠模型学习和记忆能力的分析

A Morris water maze (MWM) experiment forces experimental animals to swim and learn to find a platform hidden in the water. It is widely used in scientific ...

Nest Building Behavior as an Early Indicator of Behavioral Deficits in Mice

Nest building is an innate behavior in male and female rodents, even when raised in laboratory settings. As such, many researchers provide rodents synthetic and/or natural materials (such as twine, tissue, cotton, paper, and hay) as a gauge of their overall well-being and as an ancillary assessment to predict the possible decline in cognition. Typically, changes in nesting behaviors, such as failure to create a nest, indicate a change in health or welfare. In addition, nesting behavior is sensitive to many environmental and physiological challenges, as well as many genetic mutations underlying pathological disease states. The following protocol describes a nesting behavior paradigm that explores the usage of four types of nesting material. In addition, the protocol utilizes intraclass correlations to demonstrate that inter-rater reliability is higher when nests are constructed out of shredded paper compared to other common nesting materials such as cotton squares, paper twists, and soft cob bedding. The chosen methodology and statistical considerations (i.e., intraclass correlation) for this assay may be of interest for those conducting experiments assessing the quality of living of mice.&#160;

Here, we present a protocol to quantify nest building behavior in mice, which is known to be impaired in several neurological disorders and diseases. This protocol examines the utility of four materials and offers the opportunity to quantify the rater agreement in scoring, improving the validity and reliability of the assay.

巢穴建筑行为作为小鼠行为缺陷的早期指标

Nest building is an innate behavior in male and female rodents, even when raised in laboratory settings. As such, many researchers provide rodents ...

Chronic Post-Ischemia Pain Model for Complex Regional Pain Syndrome Type-I in Rats

Complex regional pain syndrome type-I (CRPS-I) is a neurological disease that causes severe pain among patients and remains an unresolved medical condition. However, the underlying mechanisms of CRPS-I have yet to be revealed. It is known that ischemia/reperfusion is one of the leading factors that causes CRPS-I. By means of prolonged ischemia and reperfusion of the hind limb, the rat chronic post-ischemia pain (CPIP) model has been established to mimic CRPS-I. The CPIP model has become a well-recognized animal model for studying the mechanisms of CRPS-I. This protocol describes the detailed procedures involved in the establishment of the rat model of CPIP, including anesthesia, followed by ischemia/reperfusion of the hind limb. Characteristics of the rat CPIP model are further evaluated by measuring the mechanical and thermal hypersensitivities of the hind limb as well as the nocifensive responses to acute capsaicin injection. The rat CPIP model exhibits several CRPS-I-like manifestations, including hind limb edema and hyperemia in the early stage after establishment, persistent thermal and mechanical hypersensitivities, and increased nocifensive responses to acute capsaicin injection. These characteristics render it a suitable animal model for further investigation of the mechanisms involved in CRPS-I.

Provided here is a protocol that details steps to establish an animal model of chronic post-ischemia pain (CPIP). This is a well-recognized model mimicking human complex regional pain syndrome type-I. Mechanical and thermal hypersensitivities are further evaluated, as well as capsaicin-induced nocifensive behaviors observed in the CPIP rat model.

大鼠复杂区域疼痛综合征I型慢性缺血后疼痛模型

Complex regional pain syndrome type-I (CRPS-I) is a neurological disease that causes severe pain among patients and remains an unresolved medical ...

A Chronic Sleep Fragmentation Model using Vibrating Orbital Rotor to Induce Cognitive Deficit and Anxiety-Like Behavior in Young Wild-Type Mice

Sleep disturbance is generally common in populations as a chronic disease or a complained event. Chronic sleep disturbance is proposed to be closely linked to the pathogenesis of diseases, especially neurodegenerative diseases. We recently found that 2 months of sleep fragmentation initiated Alzheimer&#8217;s disease (AD)-like behavioral and pathological changes in young wild-type mice. Herein, we present a standardized protocol to achieve chronic sleep fragmentation (CSF). Briefly, CSF was induced by an orbital rotor vibrating at 110 rpm and operating with a repetitive cycle of 10 s-on, 110 s-off, during light-ON phase (8:00 AM&#8211;8:00 PM) continuously for up to 2 months. Impairments of spatial learning and memory, anxiety-like but not depression-like behavior in mice as consequences of CSF modeling, were evaluated with Morris water maze (MWM), Novel object recognition (NOR), Open field test (OFT) and Forced swimming test (FST). In comparison with other sleep manipulations, this protocol minimizes the handling labors and maximizes the modeling efficiency. It produces stable phenotypes in young wild-type mice and can be potentially generated for a variety of research purposes.

Presented here is a protocol for chronic sleep fragmentation (CSF) model achieved by an electrically controlled orbital rotor, which could induce confirmed cognitive deficit and anxiety-like behavior in young wild-type mice. This model can be applied to explore the pathogenesis of chronic sleep disturbance and related disorders.

使用振动轨道转子诱导年轻野生型小鼠认知缺陷和焦虑样行为的慢性睡眠碎片模型

Sleep disturbance is generally common in populations as a chronic disease or a complained event. Chronic sleep disturbance is proposed to be closely ...