上海复旦大学附属金山医院动物伦理委员会批准了这项研究。将年轻的Sprague Dawley（SD）大鼠饲养在SPF动物室中，光照从08:00到20:00，黑暗从20:00到08:00，温度控制在22°C，室内相对湿度为40%-50%。他们可以免费获得食物和水。所有动物实验操作均符合实验动物福利伦理和动物实验安全规定。
1.新生儿缺氧缺血性脑病动物模型 21
<ol><li>将出生后7天（P7）的18只Sprague Dawley（SD）大鼠雄性幼崽随机分为以下几组:假（无按摩），模型和按摩（模型+按摩组）（每组n = 6）。</li>
	<li>麻醉<ol><li>用小动物气体麻醉机施用5%七氟烷气体，调节氧气流速至4，用4%诱导分数麻醉幼鼠。通过等待小老鼠仰卧、四肢停止移动、触摸时没有反应以及肌肉放松的时间点来确认动物已完全麻醉。</li>
		<li>在麻醉的幼鼠中加入宠物眼膏和眼药水，以防止眼睛干涩。</li>
	</ol></li>
	<li>显微手术过程<ol><li>首先，用碘伏对大鼠的颈部皮肤进行一次消毒，然后用75%乙醇消毒一次（执行此程序三次），并在幼鼠的颈部中间做一个~0.5厘米长的纵向切口。被动分离右侧皮下组织，暴露颈总动脉。</li>
		<li>模型和按摩组:小心地分离颈总动脉和迷走神经（图1A）。劈开颈总动脉后，用电凝枪凝固颈总动脉。确认没有出血后，逐层缝合皮肤并再次对手术切口进行消毒（图1C）。</li>
		<li>假手术组（无脑瘫，无按摩）:仅分离颈总动脉，不对血管进行任何治疗。将皮肤逐层缝合，然后再次对手术切口进行消毒。当观察幼鼠不受限制地爬行时，将幼鼠放回母鼠旁边。</li>
	</ol></li>
	<li>手术后用碘消毒。将完成手术的大鼠置于37°C绝缘垫上，不断观察伤口是否出血，眼睛是否有炎症。</li>
	<li>将模型和按摩组的年轻大鼠置于开放的缺氧室中，观察1小时直至它们醒来。 注意:当幼鼠可以自由爬行时，它们就可以接受缺氧治疗了。</li>
	<li>缺氧治疗<ol><li>从麻醉中恢复后，将年轻大鼠置于37°C的封闭缺氧装置中，92%N 2和8%O2气体条件（2L / min）缺氧3小时。缺氧治疗后，将清醒的幼鼠放回母鼠旁边。</li>
		<li>对手术器械进行消毒，并用酒精擦拭缺氧室。手术后，用碘伏对幼鼠进行消毒，每天监测伤口是否有感染迹象，并使用无菌填充物。在SPF动物房中完成整个手术过程，并将动物保持在SPF动物房中。</li>
		<li>模型建立成功后，在不采取任何干预措施的情况下，正常提升模型组。建模完成后，对按摩组进行手动干预。 注:假手术组正常喂养，未接受缺氧治疗。</li>
	</ol></li>
	<li>将每组大鼠放在无菌和高温灭菌的床上，紧挨着母鼠。观察它们1-2小时，直到母鼠喂养后代大鼠。每天观察，检查伤口是否出血，眼睛有无炎症;观察直到伤口完全愈合。</li>
</ol>2.按摩手法参数
注意:按摩的核心技术包括按压和摩擦作为主要干预技术。在实验之前，研究人员借鉴了技术测试人员的操作，该测试人员从天津腹部按摩专家王金贵那里收集了技术。
<ol><li>将按压和摩擦力分别设置为 6 N 和 3 N，分别为22、23、24、25。</li>
</ol>3. 按摩手法
注意: 确保研究人员的拇指按压穴位稳定且持久。
<ol><li>在手术后的第二天（P8），在不打扰母鼠的情况下将幼鼠带出笼子。在按摩组中，从出生第8天到第 40天按摩 大鼠。</li>
	<li>在按摩幼鼠之前，轻轻揉搓双手，使它们升温至与幼鼠皮肤几乎相同的温度。</li>
	<li>按照以下按摩顺序:肢体穴位 → 腹部穴位 →腹部。</li>
	<li>将幼鼠置于左手掌心，用四根手指的线面从头到尾轻轻触摸幼鼠的背部1分钟，使其安静。</li>
	<li>执行以下摩擦步骤。<ol><li>来回揉搓右手拇指和食指的腹部。</li>
		<li>从近端到远端摩擦前肢~30 秒，每次按曲池 （LI11）、外观 （SJ5） 和和谷 （LI4） 点 10 次，持续 1 秒。</li>
		<li>揉搓后腿，从近端到远端~30秒，并按杨灵拳（SP9），祖三里（ST36），三音娇（SP6）和太冲（LR3）点10次，每次1秒，总周期~5分钟。</li>
	</ol></li>
	<li>用右拇指按中湾（RN12）、天舒（ST25）、关源（CV4）和启海（CV6）。按压每个穴位 10 次，每次 ~1 秒，持续 1 分钟。</li>
	<li>保持幼鼠俯卧，用右手拇指按压并揉搓腹部的中湾点。以幼鼠的肚脐为中心，用拇指顺时针120x / min移动~5分钟。</li>
	<li>最后，用右手四根手指的线面轻轻触摸幼鼠的头部到尾巴，使其安静下来，然后将其放回笼子里，让母鼠喂食。</li>
</ol>4.穴位定位
注意:对于2周内出生的大鼠，大鼠的体重和长度与小鼠相同。因此，我们在相关文献中使用小鼠按摩方法对大鼠在8-15天内对大鼠进行按摩24。
<ol><li>两周至40天后，使用以下穴位定期进行大鼠按摩治疗大鼠:LI11，SJ5，LI4，SP9，ST36，SP6，LR3，RN12，ST25，CV4和CV626。 注:有关穴位的具体位置，请参阅 表1 和其他地方26 和 图2 中大鼠穴位定位的详细说明。</li>
</ol>5. 体重和行为测试
<ol><li>每天称量P7至P40的幼鼠。</li>
	<li>在P37至P40的平衡木上进行行为实验。使用平衡木（长35cm，宽1.5cm，高100cm）测试年轻大鼠的运动平衡功能。<ol><li>将小老鼠放在平衡木上，让它跑进平衡木另一端的黑盒子里。</li>
		<li>在P37-P39上对幼鼠进行平衡木训练，每天3次，并在P40上进行测试。记录三组幼鼠通过平衡木的时间以及它们的后肢滑动的次数（图1D）。 注意:使用二氧化碳窒息法对大鼠实施安乐死。</li>
	</ol></li>
</ol>

中医按摩技术:增强脑瘫大鼠的运动功能

作者没有要披露的利益冲突。

新生大鼠缺血缺氧模型是研究脑瘫最经典的模型27,28,29,30。我们从第8天到第40天对大鼠进行干预，这大约相当于从3个月到11.5岁的人类进行干预，这是人类生长发育的黄金期31。结果表明，按摩是改善脑瘫功能障碍的有效方法。先前的动物实验表明，按摩可以减少炎症并改善CP14,32年轻大鼠的运动和认知功能。一些临床研究表明，背部和膀胱经络按摩可改善 CP33 儿童的运动功能。这些结果还表明，按摩可以缩短脑瘫幼鼠通过平衡木的时间，减少后腿滑倒的次数。由此可见，腹部按摩可以改善CP幼鼠的运动功能。
中医认为，脑瘫属于五迟、五软症状，先天性肾气虚为根本因素，后天性脾胃无力为促因素。肾掌骨，生骨髓，使肾气充沛时，骨髓变得充沛，智力保持正常。脾控制着肌肉和四肢，所以如果脾脏健康，肌肉就会强壮，关节就会自由活动。因此，有必要补充脾脏，强肾，改善胃肠功能，活动四肢关节，改善功能。首先，中医理论认为，腹部是一切疾病的根源。腹部按摩可以上下清清，分阴分阳，除旧除新，丰富五脏，驱除外邪，消除内疾。其次，研究表明，脑瘫患儿体重较轻，容易营养不良，包括摄入量、能量或营养状况过多或不平衡34,35,36,37。生长迟缓和营养不良在脑瘫患儿中很常见，与更严重的运动功能障碍直接相关37.营养不良的风险与CP34,38,39患者的预后密切相关。目前，腹部按摩的主要研究领域是改善功能性消化不良、便秘、腹泻和婴儿厌食症等胃肠道症状33,40,41,42。最终，大量研究表明，刺激中湾、天舒、观原和七海可以治疗胃肠道疾病，促进吸收，改善营养状况43,44,45。腹穴可以治疗胃肠道相关疾病，梳理中枢充气机制，补充初气之力，促进气血生化。因此，我们将腹部按摩引入实验方法，以改善CP年轻大鼠的运动功能障碍。
腹部按摩以腹穴结合，肢体穴位为辅，如曲池、足三里等。四肢穴位的外侧支和关节与内脏相连，采用腹部按摩，以达到最佳治疗效果。一方面，实验动物研究表明，对缺氧缺血性脑瘫新生大鼠进行针灸祖三里可以改善CP46幼鼠的功能。另一方面，临床研究表明，在和谷47等穴位针灸可有效促进脑瘫的发展。 曲池和足三里是手经阳明大肠的穴位48 和足阳明经的生长抑素49，是我国脑卒中临床治疗改善肢体麻痹最常用的穴位50，51.此外，曲池和祖三里分别是大肠经和胃经的汇合点。从这一点开始，经络气到达脏腑，并与上下部分以及经络和侧支相连。此外，针灸加推拿在临床研究结果中显示出更好的改善儿童脑瘫效果52。综上所述，中医治疗可以帮助改善脑瘫患者的功能障碍。
尽管关于按摩如何改善脑瘫的研究很多，但按摩技术的多样性已成为一个巨大的挑战。按摩学校很多，实力、方向等因素的差异带来了诸多挑战。我们学习了多种按摩方法，探索了一套简单易行的按摩手法。但是，我们无法模拟各种按摩技术。其次，按摩的时间是幼鼠发育的重要阶段。它的重量和各种身体系统的快速生长导致我们不同阶段的大鼠的手指力量不断变化。必须继续努力研究按摩的稳定性。

脑性瘫痪（CP）是一种致残率和发病率高的疾病。脑性瘫痪的流行病学调查显示，每1000名婴儿中就有3-5名婴儿患有脑瘫1。脑瘫的典型症状是持续性中枢运动和姿势发育障碍2、感觉活动受限3、知觉、认知4、沟通5、行为障碍、癫痫和继发性肌肉骨骼问题1。到目前为止，这种终生致残的疾病还没有治愈方法。目前的治疗方法包括运动疗法6、水疗7、功能性电刺激8、肌肉外骨骼9、脐带血细胞疗法10、肉毒杆菌毒素注射和手术11。然而，临床康复耗时长，耗费精力大，康复需要患者的积极配合，难以在动物模型中复制。手术有局限性、适应证和不可逆性。口服药物治疗脑瘫患儿运动功能效果差12.肌肉外骨骼治疗和脐带血治疗经济成本高，难以负担，因此找到合适的治疗策略至关重要。
按摩是一种非侵入性、无副作用、被广泛接受、具有成本效益的儿童治疗方法。临床13 和动物实验14 均已证明可以改善脑瘫功能障碍。 中医认为按摩可以通过调节器官和疏通经络来改善脑瘫功能障碍15.按摩作为一种有效的中医治疗方法，需要现代医学研究方法解释其作用机理，才能推广到世界。然而，在脑瘫患儿相关机制的研究中，存在客观的研究局限性和伦理局限性。因此，研究改善动物脑瘫的相关按摩机制更加方便。虽然动物与儿童不同，但研究按摩对动物的影响需要坚持中医的基本理论。从临床和实验动物学的角度来看，我们将把这项技术从人体复制到动物身上，研究按摩的效果，这将有助于促进按摩疗法的应用。
首先，中医认为脑瘫属于"五迟"和"五软"的症状16。脑瘫的病因是先天性胎功能障碍、肝肾虚、气气不足或后天性营养缺乏15。肾脏主导着人体器官和组织的生长、发育或再生和修复，这些物质对身体的生长、发育、组织修复和复活至关重要。肾脏的精气是生长发育的基础。肾精不足会导致骨髓衰竭、气血紊乱、身体内脏、肌肉、骨骼缺乏营养。脑瘫患儿容易出现生长发育迟缓、四肢关节活动度下降和肌肉紧张增加。此外，长期患病可导致过度虚血瘀，引起经络阻塞，导致智力障碍、反应迟钝、运动障碍、语言模糊、吞咽无力、流口水等症状，如肢体运动障碍。
脾掌管血液循环和转化，是气血生化之源，是后天能力的基础。连接脏腑、胳膊和腿的人体经络是气血循环、上下沟通、调节阴阳的通路。按摩仁经可以增强肾功能，调气调血。在按摩手法中，利用足太阴脾经、足阳明胃经、手阳明大肠经穴位，改善脾胃功能，有助于促进气血循环。按摩利用经络、特定部位、穴位对儿童身体进行良性的物理刺激，引起由表面到内、由外到内的反应性变化，从而改善局部和整体血液循环，恢复大脑的正常兴奋性和抑制力，达到内脏和组织血液功能的相应平衡。根据这个理论，我们主要对腹部、中湾（RN12）、天舒（ST25）、观元（CV4）和气海（CV6）等穴位进行按摩。腹部按摩可以改善CP17,18,19患儿的临床症状，极有可能通过脑肠通路促进脑损伤修复20。因为脾胃是产后发育的基础，所以可以通过健脾、益胃、补气补血来促进脑损伤恢复。
我们在大鼠身上复制了这种技术，以尽可能模拟中医对脑瘫的干预，发现腹部按摩可以改善脑瘫大鼠的功能障碍。综上所述，我们提出了一套研究腹部按摩的方法，以研究通过腹部按摩改善CP的相关机制。由于按摩流派多，按摩机理研究难度大，因此需要规范按摩方法。

<table><tbody><tr><td>Animal anesthesia machine</td><td>RWD OF SCIENCE</td><td>R550</td><td></td></tr><tr><td>Experimental gloves</td><td>Sri Trang Gloves (Thailand) Co.,Ltd</td><td>0169</td><td></td></tr><tr><td>Isoflurane</td><td>RWD OF SCIENCE</td><td>R510-22-10</td><td></td></tr><tr><td>Rat</td><td>Shanghai Jihui Experimental Animal Breeding Co., Ltd.</td><td></td><td></td></tr><tr><td>YF-3 manipulation tester</td><td>Developed jointly by Shanghai Fudan University and Shanghai University of Traditional Chinese Medicine</td><td></td><td>Patent number of the People&#039;s Republic of China (ZL200710187403.1)</td></tr></tbody></table>

abdominal massage to improve motor dysfunction in rats with cerebral palsy

脑性瘫痪（CP）是一种致残率和发病率高的疾病。脑瘫的临床症状为运动功能障碍和姿势发育异常，常伴有认知障碍。按摩是一种传统的中医疗法，可以协调藏符，调节气血，使脏腑工作更顺畅，镇静阴阳。此外，它已成为临床上CP的有效方法。本文总结了一套简单规范的脑瘫幼鼠按摩操作方法，简单易懂。操作步骤如下:首先，按摩四个肢体穴位，包括曲池（LI11）和足三里（ST36）;二是按摩腹部穴位中婉（RN12）、天舒（ST25）、观元（CV4）、启海（CV6）;最后，按摩大鼠的腹部。这套按摩方法大大改善了CP年轻大鼠的运动功能，简单、规范、易操作。我们在动物模型中对这套操作方法进行了调整，以促进按摩的国际化和标准化。

在完成缺血缺氧建模27、28、29、30（实验在SPF动物室完成）后，取存活48 h后幼鼠的大脑寻找水肿、液化等现象，如图3所示。比较假手术组和按摩组幼鼠体重变化情况与模型组（附表S1）。模型组体重增加速度慢于按摩组（图4），说明按摩可改善CP幼鼠的生长发育。平衡木测试结果显示，模型组穿过平衡木的时间比假平衡木组和按摩木组长（图5A和补充表S2）。
此外，与假手术组相比，模型组的后肢滑脱次数显著增加（补充表S3）。相比之下，与模型组相比，按摩组的后肢滑倒次数显着减少（图5B 和 补充表S3）。按摩干预显著改善了CP大鼠的生长和运动功能。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/65625/65625fig01.jpg"> 图1:动物建模过程和相关行为测试 。 （A） 建模过程中迷走神经的剥离。（B）按摩5周龄大鼠。（C）缺血完成后的大鼠。（四）平衡木试验。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/65625/65625fig01large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/65625/65625fig02.jpg"> 图2:部分与腹部按摩相关的穴位展示26. <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/65625/65625fig02large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/65625/65625fig03.jpg"> 图 3:模型建立后 48 小时后一侧大脑的水肿。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/65625/65625fig03large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/65625/65625fig04.jpg"> 图4:按摩疗法对体重的影响。 从P7到P40测量大鼠幼崽（n = 6）的平均体重，对P7进行缺氧缺血干预。HIE模型后，模型组体重增长率显著下降，按摩疗法显著改善（p < 0.05）。数据以标准差±平均值表示;#p < 0.05 按摩组与模型组相比。*P < 0.001 模型组与假模型组相比。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/65625/65625fig04large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/65625/65625fig05.jpg"> 图5:按摩治疗对脑瘫大鼠运动功能障碍的影响。 （A）大鼠穿过光束所需的时间（n = 6）。（B）大鼠后肢滑脱的次数。根据图4A，B的结果，按摩组的运动性能明显优于模型组（p < 0.05）。数据以标准差±平均值表示;*p < 0.001 模型组与假手术组相比。#p < 0.01 按摩组与模型组相比。**p < 0.0001 模型组与假手术组相比。##p < 0.0001 按摩组与模型组相比。<a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/65625/65625fig05large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody><tr><td>李11</td>
			<td colspan="5">桡侧腕伸肌的内凹陷和</td>
		</tr><tr><td>SJ5型</td>
			<td colspan="5">肘部外凹陷 在虚形远端 5/6 点的桡尺凹陷处</td>
		</tr><tr><td>李4</td>
			<td colspan="5">连接腕部和肘部的线，伸肌后方 趾状肌腱</td>
		</tr><tr><td>SP9型</td>
			<td colspan="5">在第一背骨间，内侧至第二骨中间 掌骨，在前掌</td>
		</tr><tr><td>ST36型</td>
			<td colspan="5">在胫骨内侧髁下方的凹陷处， 胫骨和腓肠肌之间</td>
		</tr><tr><td>SP6型</td>
			<td colspan="5">在从 ST35 到前部的线上的近端第五点 脚踝皱褶侧</td>
		</tr><tr><td>磁共振成像</td>
			<td colspan="5">在假想线上远端五分之一的近端端点 连接SP9和胫腓骨内踝</td>
		</tr><tr><td>CV4型</td>
			<td colspan="5">在第一和第二凹陷之间的凹陷的近端 跖骨</td>
		</tr><tr><td>ST25型</td>
			<td colspan="5">在腹侧中线向下 3/5 处连接 脐部至耻骨结节</td>
		</tr><tr><td>CV6型</td>
			<td colspan="5">脐外侧，从脐到线的中点 在 3/10 的腹侧中线连接 脐部至耻骨结节</td>
		</tr></tbody></table>表1:各种穴位的解剖位置26. 
补充表S1:所有三组大鼠体重的原始数据。<a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/65625/65625_Supp%20Table%20S1%20weight.xls" target="_blank">请点击这里下载此文件。</a>
补充表S2:所有三组大鼠越过平衡木的时间的原始数据。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/65625/65625_Supp%20Table%20S2%20Time%20%20to%20cross%20%20the%20%20beam.xls" target="_blank">请点击这里下载此文件。</a>
补充表S3:三组大鼠后肢滑脱的原始数据。<a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/65625/65625_Supp%20Table%20S3%20Number%20%20of%20slips%20of%20the%20hind%20%20limbs.xls" target="_blank">请点击这里下载此文件。</a>

Watch this Scientific Journal Video about Exploring the Potential of Massage Therapy in Cerebral Palsy Using Animal Models at JoVE.com

作者聚焦:使用动物模型探索脑瘫按摩疗法的潜力 

在这里，我们在脑瘫大鼠模型中提出了一组按摩操作，可以显着改善脑瘫大鼠的运动功能。

腹部按摩改善脑瘫大鼠运动功能障碍

Cerebral palsy (CP) is a disease with a high disability rate and morbidity. The clinical symptoms of cerebral palsy are motor dysfunction and abnormal ...

我们概述了一种新颖的按摩技术，以缓解受伤的新大鼠的脑瘫症状。目前，关于使用按摩疗法改善脑瘫症状的动物研究很少。然而，我们发现年轻大鼠腹部按摩的有希望的结果可能会提高它们的操纵能力。关于按摩如何改善脑瘫的研究有限。然而，我们已经建立了动物模型和按摩技术，可以促进未来对脑瘫按摩机制的研究。推拿疗法对脑瘫至关重要。研究动物按摩技术稳定后的相关机制可以推进研究。中式按摩可以对脑瘫的生长发育产生积极影响，因此我们将重点介绍中式按摩在脑瘫中的应用。

abdominal-massage-to-improve-motor-dysfunction-rats-with-cerebral

Nanyang Technological University

Research

JoVE Journal

Neuroscience

Abdominal Massage to Improve Motor Dysfunction in Rats with Cerebral Palsy

1.9K 次观看.  Yunnan University of Traditional Chinese Medicine. 我们概述了一种新颖的按摩技术，以缓解受伤的新大鼠的脑瘫症状。目前，关于使用按摩疗法改善脑瘫症状的动物研究很少。然而，我们发现年轻大鼠腹部按摩的有希望的结果可能会提高它们的操纵能力。关于按摩如何改善脑瘫的研究有限。然而，我们已经建立了动物模型和按摩技术，可以促进未来对脑瘫按摩机制的研究。推拿疗法对脑瘫至关重要。研究动物按摩...

视频: 作者聚焦:使用动物模型探索脑瘫按摩疗法的潜力 

Dual Electrophysiological Recordings of Synaptically-evoked Astroglial and Neuronal Responses in Acute Hippocampal Slices

Astrocytes form together with neurons tripartite synapses, where they integrate and modulate neuronal activity. Indeed, astrocytes sense neuronal inputs through activation of their ion channels and neurotransmitter receptors, and process information in part through activity-dependent release of gliotransmitters. Furthermore, astrocytes constitute the main uptake system for glutamate, contribute to potassium spatial buffering, as well as to GABA clearance. These cells therefore constantly monitor synaptic activity, and are thereby sensitive indicators for alterations in synaptically-released glutamate, GABA and extracellular potassium levels. Additionally, alterations in astroglial uptake activity or buffering capacity can have severe effects on neuronal functions, and might be overlooked when characterizing physiopathological situations or knockout mice. Dual recording of neuronal and astroglial activities is therefore an important method to study alterations in synaptic strength associated to concomitant changes in astroglial uptake and buffering capacities. Here we describe how to prepare hippocampal slices, how to identify stratum radiatum astrocytes, and how to record simultaneously neuronal and astroglial electrophysiological responses. Furthermore, we describe how to isolate pharmacologically the synaptically-evoked astroglial currents.

The preparation of acute brain slices from isolated hippocampi, as well as the simultaneous electrophysiological recordings of astrocytes and neurons in stratum radiatum during stimulation of schaffer collaterals is described. The pharmacological isolation of astroglial potassium and glutamate transporter currents is demonstrated.

双突触诱发的星形胶质细胞和神经元的反应，在急性海马脑片电生理记录

Astrocytes form together with neurons tripartite synapses, where they integrate and modulate neuronal activity. Indeed, astrocytes sense neuronal inputs ...

科研

神经科学

Utilizing 3D Printing Technology to Merge MRI with Histology: A Protocol for Brain Sectioning

Magnetic resonance imaging (MRI) allows for the delineation between normal and abnormal tissue on a macroscopic scale, sampling an entire tissue volume three-dimensionally. While MRI is an extremely sensitive tool for detecting tissue abnormalities, association of signal changes with an underlying pathological process is usually not straightforward. In the central nervous system, for example, inflammation, demyelination, axonal damage, gliosis, and neuronal death may all induce similar findings on MRI. As such, interpretation of MRI scans depends on the context, and radiological-histopathological correlation is therefore of the utmost importance. Unfortunately, traditional pathological sectioning of brain tissue is often imprecise and inconsistent, thus complicating the comparison between histology sections and MRI. This article presents novel methodology for accurately sectioning primate brain tissues and thus allowing precise matching between histology and MRI. The detailed protocol described in this article will assist investigators in applying this method, which relies on the creation of 3D printed brain slicers. Slightly modified, it can be easily implemented for brains of other species, including humans.

The overall goal of this protocol is to accurately align magnetic resonance imaging (MRI) image volumes with histology sections via the creation of customized 3D-printed brain holders and slicer boxes.

利用3D打印技术来合并MRI与组织学:一个大脑切片协议

Magnetic resonance imaging (MRI) allows for the delineation between normal and abnormal tissue on a macroscopic scale, sampling an entire tissue volume ...

The Use of Trace Eyeblink Classical Conditioning to Assess Hippocampal Dysfunction in a Rat Model of Fetal Alcohol Spectrum Disorders

Neonatal rats were administered a relatively high concentration of ethyl alcohol (11.9% v/v) during postnatal days 4-9, a time when the fetal brain undergoes rapid organizational change and is similar to accelerated brain changes that occur during the third trimester in humans. This model of fetal alcohol spectrum disorders (FASDs) produces severe brain damage, mimicking the amount and pattern of binge-drinking that occurs in some pregnant alcoholic mothers. We describe the use of trace eyeblink classical conditioning (ECC), a higher-order variant of associative learning, to assess long-term hippocampal dysfunction that is typically seen in alcohol-exposed adult offspring. At 90 days of age, rodents were surgically prepared with recording and stimulating electrodes, which measured electromyographic (EMG) blink activity from the left eyelid muscle and delivered mild shock posterior to the left eye, respectively. After a 5 day recovery period, they underwent 6 sessions of trace ECC to determine associative learning differences between alcohol-exposed and control rats. Trace ECC is one of many possible ECC procedures that can be easily modified using the same equipment and software, so that different neural systems can be assessed. ECC procedures in general, can be used as diagnostic tools for detecting neural pathology in different brain systems and different conditions that insult the brain.

Trace eyeblink classical conditioning (ECC) was used to assess hippocampal-dependent associative learning in adult rats that were administered a high concentration (11.9% v/v) of alcohol during early neonatal brain development. In general, ECC procedures are sound diagnostic tools for detecting brain dysfunction across many psychological and biomedical settings.

使用的跟踪经典条件反射，评估胎儿酒精谱系障碍大鼠模型海马功能障碍

Neonatal rats were administered a relatively high concentration of ethyl alcohol (11.9% v/v) during postnatal days 4-9, a time when the fetal brain ...

A Rat Model of Mild Intrauterine Hypoperfusion with Microcoil Stenosis

Intrauterine hypoperfusion/ischemia is one of the major causes of intrauterine/fetal growth restriction, preterm birth, and low birth weight. Most studies of this phenomenon have been performed in either models with severe intrauterine ischemia or models with gradient degree of intrauterine hypoperfusion. No study has been performed in a model on uniform mild intrauterine hypoperfusion (MIUH). Two models have been used for studies of MIUH: a model based on suture ligation of either side of the arterial arcade formed with the uterine and ovarian arteries, and a transient model based on clipping the bilateral ovarian arteries and aorta having patency. Those two rodent models of MIUH have some limitations, e.g., not all fetuses are subjected to MIUH, depending on their position in the uterine horn. In our MIUH model, all fetuses are subjected to a comparable level of intrauterine hypoperfusion. MIUH was achieved by mild stenosis of all four arteries feeding the uterus, i.e., the bilateral uterine and ovarian arteries.
Arterial stenosis was induced by metal microcoils wrapped around the feeding arteries. Producing arterial stenosis with microcoils allowed us to control, optimize, and reproduce decreased blood flow with very little inter-animal variability and a low mortality rate, thus enabling accurate evaluation. When microcoils with an inner diameter of 0.24 mm were used, the blood flow in both the placenta and fetus was mildly decreased (approximately 30% from the pre-stenosis level in the placenta). The offspring of our MIUH model clearly demonstrates long-lasting alterations in neurological, neuroanatomical and behavioral test results.

Mild intrauterine hypoperfusion was produced by artery stenosis with metal microcoils wrapped around the uterine and ovarian arteries in rats at embryonic day 17. This procedure produced prenatal hypoperfusion and intrauterine growth restriction.

轻度宫内灌注圈狭窄的大鼠模型

Intrauterine hypoperfusion/ischemia is one of the major causes of intrauterine/fetal growth restriction, preterm birth, and low birth weight. Most studies ...

Sub-acute Cerebral Microhemorrhages Induced by Lipopolysaccharide Injection in Rats

Cerebral microhemorrhages (CMHs) are common in aged patients and are correlated to various neuropsychiatric disorders. The etiology of CMHs is complex, and neuroinflammation is often observed as a co-occurrence. Here, we describe a sub-acute CMHs rat model induced by lipopolysaccharide (LPS) injection, as well as a method for detecting CMHs. Systemic LPS injection is relatively simple, economical, and cost-effective. One major advantage of LPS injection is its stability to induce inflammation. CMHs caused by LPS injection could be detected by gross observation, hematoxylin and eosin (HE) staining, Perl&#39;s Prussian staining, Evans blue (EB) double-labeling, and magnetic resonance imaging-susceptibility weighted imaging (MRI-SWI) technology. Finally, other methods of developing CMHs animal models, including their advantages and/or disadvantages, are also discussed in this report.

We present a protocol to induce and detect CMHs caused by LPS injection in Sprague-Dawley rats, which may be utilized in future research investigations on the pathogenesis of CMHs.

脂多糖注射液致大鼠亚急性脑 Microhemorrhages

Cerebral microhemorrhages (CMHs) are common in aged patients and are correlated to various neuropsychiatric disorders. The etiology of CMHs is complex, ...

Single Synapse Indicators of Glutamate Release and Uptake in Acute Brain Slices from Normal and Huntington Mice

Synapses are highly compartmentalized functional units that operate independently on each other. In Huntington&#39;s disease (HD) and other neurodegenerative disorders, this independence might be compromised due to insufficient glutamate clearance and the resulting spill-in and spill-out effects. Altered astrocytic coverage of the presynaptic terminals and/or dendritic spines as well as a reduced size of glutamate transporter clusters at glutamate release sites have been implicated in the pathogenesis of diseases resulting in symptoms of dys-/hyperkinesia. However, the mechanisms leading to the dysfunction of glutamatergic synapses in HD are not well understood. Improving and applying synapse imaging we have obtained data shedding new light on the mechanisms impeding the initiation of movements. Here, we describe the principle elements of a relatively inexpensive approach to achieve single synapse resolution by using the new genetically encoded ultrafast glutamate sensor iGluu, wide-field optics, a scientific CMOS (sCMOS) camera, a 473 nm laser and a laser positioning system to evaluate the state of corticostriatal synapses in acute slices from age appropriate healthy or diseased mice. Glutamate transients were constructed from single or multiple pixels to obtain estimates of i) glutamate release based on the maximal elevation of the glutamate concentration [Glu] next to the active zone and ii) glutamate uptake as reflected in the time constant of decay (TauD) of the perisynaptic [Glu]. Differences in the resting bouton size and contrasting patterns of short-term plasticity served as criteria for the identification of corticostriatal terminals as belonging to the intratelencephalic (IT) or the pyramidal tract (PT) pathway. Using these methods, we discovered that in symptomatic HD mice ~40% of PT-type corticostriatal synapses exhibited insufficient glutamate clearance, suggesting that these synapses might be at risk to excitotoxic damage. The results underline the usefulness of TauD as a biomarker of dysfunctional synapses in Huntington mice with a hypokinetic phenotype.

We present a protocol to evaluate the balance between glutamate release and clearance at single corticostriatal glutamatergic synapses in acute slices from adult mice. This protocol uses the fluorescent sensor iGluu for glutamate detection, a sCMOS camera for signal acquisition and a device for focal laser illumination.

正常小鼠和亨廷顿小鼠急性脑切片中谷氨酸释放和服用的单突触指标

Synapses are highly compartmentalized functional units that operate independently on each other. In Huntington&#39;s disease (HD) and other ...

Developmental Enhancement in Cerebral Palsy Rats via Spinal Manipulations

Enhancing the Development and Growth of Infant Cerebral Palsy Rats Using Selective Spinal Manipulations

Cerebral palsy (CP) is a refractory pediatric disease with a high prevalence, high disability rate, and difficult treatment. A variety of treatments are currently used for CP. The treatment involves drug and non-drug therapy. Traditional Chinese medicine external therapy is a very distinctive treatment method in non-drug therapy. As one of the external therapies of traditional Chinese medicine, massage is used in treating cerebral palsy and has good efficacy, small side effects, and strong operability. As a part of TCM external therapy, selective spinal manipulation can effectively promote the growth and development of infant rats with cerebral palsy.The operation was mainly divided into four steps: first, the rubbing method was applied to the spine and both sides of the spine for 1 min. The pressing and kneading method was applied to the spine for 5 min, and the muscles on both sides of the spine for 5 min. Second, pressing and kneading the sensitive local acupoints in the spine for 2 min were performed. Thirdly, the affected limb was treated by twisting method for 1 min. Fourth, the rubbing method was applied to a midline from the forehead to the back of the brain for 1 min. This study aimed to use selective spinal manipulation to treat infant rats with cerebral palsy. The weight, Rotarod test, Foot-fault score, and growth hormone of infant rats with cerebral palsy were detected to understand the effect of selective spinal manipulation on the growth and development of infant rats with cerebral palsy. The results showed that it can promote weight gain, improve balance ability and motor function, promote growth and development of infant cerebral palsy rats, promote growth hormone secretion, and increase the temperature of sensitive parts of the back.

This study illustrates the effect of selective spinal manipulation on the growth and development of infant rats with cerebral palsy, emphasizing the specific procedure and standardized protocol. Body weight measurement, Rotarod test, Foot-fault score, other behavioral tests, and growth hormone detection were performed to evaluate the protocol.

Cerebral palsy (CP) is a refractory pediatric disease with a high prevalence, high disability rate, and difficult treatment. A variety of treatments are ...

基于 Ding's Roll 操作治疗大鼠模型肌肉损伤的按摩器

Mimicking Ding's Roll Method on Notexin-Induced Muscle Injury in Rats

Ding's roll method is one of the most commonly used manipulations in traditional Chinese massage (Tuina) clinics and one of the most influential contemporary Tuina manipulations in China. It is based on the traditional rolling method commonly used in the one finger Zen genre and named Ding's roll method. Due to its anti-inflammatory and blood circulation-promoting effects, Ding's rolling method has sound therapeutic effects on myopathy. Because of the large area of force applied to human skin, Ding's roll method is challenging to perform on experimental animals with small skin areas, such as rats and rabbits. Additionally, the strength of Tuina applied to the human body differs from that applied to experimental animals, so it may happen that the strength is too high or too low to achieve the therapeutic effect of Tuina during the experiment. This experiment aims to create a simple massager suitable for rats based on Ding's rolling manipulation parameters (strength, frequency, Tuina duration). The device can standardize manipulation in animal experiments and reduce the variation in Tuina force applied to different animals due to subjective factors. A rat model of notexin-induced skeletal muscle injury was established, and plasma injury markers creatine kinase (CK) and fatty acid binding protein 3 (FABP3) were used to assess the therapeutic effect of Tuina on skeletal muscle injury. The results showed that this Tuina massager could reduce the levels of CK and FABP3 expression and slow down the degree of skeletal muscle injury. Therefore, the Tuina massager described here, mimicking Ding's roll method, contributes to standardizing Tuina manipulation in experimental research and is of great help for subsequent research on the molecular mechanism of Tuina for myopathy.

作者聚焦：使用自制按摩器对大鼠进行推拿按摩 

This protocol describes a simple device that mimics Ding's roll method, establishes a rat model of skeletal muscle injury, and uses hematoxylin-eosin staining to observe the pathology of damaged tissue and enzyme-linked immunosorbent assay to detect changes in serum damage markers.

腹部按摩改善脑瘫大鼠运动功能障碍

副本

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此视频中的章节

双突触诱发的星形胶质细胞和神经元的反应，在急性海马脑片电生理记录

利用3D打印技术来合并MRI与组织学:一个大脑切片协议

使用的跟踪经典条件反射，评估胎儿酒精谱系障碍大鼠模型海马功能障碍

轻度宫内灌注圈狭窄的大鼠模型

脂多糖注射液致大鼠亚急性脑 Microhemorrhages

正常小鼠和亨廷顿小鼠急性脑切片中谷氨酸释放和服用的单突触指标

Enhancing the Development and Growth of Infant Cerebral Palsy Rats Using Selective Spinal Manipulations

模仿丁氏滚动法治疗大鼠肌损伤

模仿丁氏滚动法治疗大鼠肌损伤

模仿丁氏滚动法治疗大鼠肌损伤