Name: dural stimulation and periorbital von frey testing in mice as a preclinical model of headache
Uploaded: 2021-07-29T15:00:00
Description: Watch this Scientific Journal Video about Dural Stimulation and Periorbital von Frey Testing in Mice As a Preclinical Model of Headache at JoVE.com

这项研究得到了美国国立卫生研究院的支持（NS104200和NS072204至GD）。

所有程序都是在德克萨斯大学达拉斯分校动物护理和使用委员会的事先批准下进行的。本研究使用ICR（CD-1）（30-35g）和C57 / BL6（25-30g）小鼠，年龄在6-8周。
1. 硬脑膜浸泡器
<ol><li>通过修改市售的用于单侧注射的市售内套管和注入器来创建小鼠注射器/注射器，该盖子可调节并插入/延伸到内径（I.D.）为0.18 mm，外径（O.D.）为0.35 mm的28 G导向套管下方（图1A</strong...

<ol>
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P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Migraine+affects+1+in+10+people+worldwide+featuring+recent+rise:+A+systematic+review+and+meta-analysis+of+community-based+studies+involving+6+million+participants.">Migraine affects 1 in 10 people worldwide featuring recent rise: A systematic review and meta-analysis of community-based studies involving 6 million participants.</a> Journal of the Neurological Sciences. 372, 307-315 (2017).</li><li>Burch, R. C., Loder, S., Loder, E., Smitherman, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+prevalence+and+burden+of+migraine+and+severe+headache+in+the+United+States:+updated+statistics+from+government+health+surveillance+studies.">The prevalence and burden of migraine and severe headache in the United States: updated statistics from government health surveillance studies.</a> Headache. 55 (1), 21-34 (2015).</li><li>Ashina, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Migraine.">Migraine.</a> New England Journal of Medicine. 383 (19), 1866-1876 (2020).</li><li>Ashina, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Migraine:+integrated+approaches+to+clinical+management+and+emerging+treatments.">Migraine: integrated approaches to clinical management and emerging treatments.</a> Lancet. 397 (10283), 1505-1518 (2021).</li><li>Jacobs, B., Dussor, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurovascular+contributions+to+migraine:+Moving+beyond+vasodilation.">Neurovascular contributions to migraine: Moving beyond vasodilation.</a> Neuroscience. 338, 130-144 (2016).</li><li>Koyuncu Irmak, D., Kilinc, E., Tore, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Shared+Fate+of+Meningeal+Mast+Cells+and+Sensory+Neurons+in+Migraine.">Shared Fate of Meningeal Mast Cells and Sensory Neurons in Migraine.</a> Frontiers in Cellular Neuroscience. 13, 136 (2019).</li><li>Levy, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Migraine+pain,+meningeal+inflammation,+and+mast+cells.">Migraine pain, meningeal inflammation, and mast cells.</a> Current Pain and Headache Reports. 13 (3), 237-240 (2009).</li><li>Levy, D., Labastida-Ramirez, A., MaassenVanDenBrink, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Current+understanding+of+meningeal+and+cerebral+vascular+function+underlying+migraine+headache.">Current understanding of meningeal and cerebral vascular function underlying migraine headache.</a> Cephalalgia. 39 (13), 1606-1622 (2019).</li><li>Phebus, L. A., Johnson, K. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dural+inflammation+model+of+migraine+pain.">Dural inflammation model of migraine pain.</a> Current Protocols in Neuroscience. , (2001).</li><li>Fried, N. T., Maxwell, C. R., Elliott, M. B., Oshinsky, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Region-specific+disruption+of+the+blood-brain+barrier+following+repeated+inflammatory+dural+stimulation+in+a+rat+model+of+chronic+trigeminal+allodynia.">Region-specific disruption of the blood-brain barrier following repeated inflammatory dural stimulation in a rat model of chronic trigeminal allodynia.</a> Cephalalgia. 38 (4), 674-689 (2018).</li><li>Avona, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dural+calcitonin+gene-related+peptide+produces+female-specific+responses+in+rodent+migraine+models.">Dural calcitonin gene-related peptide produces female-specific responses in rodent migraine models.</a> The Journal of Neuroscience. 39 (22), 4323-4331 (2019).</li><li>Burgos-Vega, C. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-invasive+dural+stimulation+in+mice:+A+novel+preclinical+model+of+migraine.">Non-invasive dural stimulation in mice: A novel preclinical model of migraine.</a> Cephalalgia. 39 (1), 123-134 (2019).</li><li>Avona, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Meningeal+CGRP-Prolactin+interaction+evokes+female-specific+migraine+behavior.">Meningeal CGRP-Prolactin interaction evokes female-specific migraine behavior.</a> Annals of Neurology. 89 (6), 1129-1144 (2021).</li><li>Deuis, J. R., Dvorakova, L. S., Vetter, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methods+used+to+evaluate+pain+behaviors+in+rodents.">Methods used to evaluate pain behaviors in rodents.</a> Frontiers in Molecular Neuroscience. 10, 284 (2017).</li><li>Lipton, R. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cutaneous+allodynia+in+the+migraine+population.">Cutaneous allodynia in the migraine population.</a> Annals of Neurology. 63 (2), 148-158 (2008).</li><li>Goadsby, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Migraine,+allodynia,+sensitisation+and+all+of+that.">Migraine, allodynia, sensitisation and all of that.</a> European Neurology. 53, 10-16 (2005).</li><li>Langford, D. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coding+of+facial+expressions+of+pain+in+the+laboratory+mouse.">Coding of facial expressions of pain in the laboratory mouse.</a> Nature Methods. 7 (6), 447-449 (2010).</li><li>Mogil, J. S., Pang, D. S. J., Silva Dutra, G. G., Chambers, C. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+development+and+use+of+facial+grimace+scales+for+pain+measurement+in+animals.">The development and use of facial grimace scales for pain measurement in animals.</a> Neuroscience &amp; Biobehavioral Reviews. 116, 480-493 (2020).</li><li>Vuralli, D., Wattiez, A. S., Russo, A. F., Bolay, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+and+cognitive+animal+models+in+headache+research.">Behavioral and cognitive animal models in headache research.</a> The Journal of Headache and Pain. 20 (1), 11 (2019).</li><li>Mason, B. N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+migraine-like+photophobic+behavior+in+mice+by+both+peripheral+and+central+CGRP+mechanisms.">Induction of migraine-like photophobic behavior in mice by both peripheral and central CGRP mechanisms.</a> The journal of Neuroscience. 37 (1), 204-216 (2017).</li><li>Dixon, W. J., Mood, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+method+for+obtaining+and+analyzing+sensitivity+data.">A method for obtaining and analyzing sensitivity data.</a> The Journal of the American Statistical Association. 43 (241), 109-126 (1948).</li><li>Dixon, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+up-and-down+method+for+small+samples.">The up-and-down method for small samples.</a> The Journal of the American Statistical Association. 60, (1965).</li><li>Bonin, R. P., Bories, C., De Koninck, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simplified+up-down+method+(SUDO)+for+measuring+mechanical+nociception+in+rodents+using+von+Frey+filaments.">A simplified up-down method (SUDO) for measuring mechanical nociception in rodents using von Frey filaments.</a> Molecular Pain. 10, 26 (2014).</li><li>Ramachandran, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurogenic+inflammation+and+its+role+in+migraine.">Neurogenic inflammation and its role in migraine.</a> Seminars in Immunopathology. 40 (3), 301-314 (2018).</li><li>Edvinsson, L., Haanes, K. A., Warfvinge, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Does+inflammation+have+a+role+in+migraine.">Does inflammation have a role in migraine.</a> Nature Reviews Neurology. 15 (8), 483-490 (2019).</li><li>Stokely, M. E., Orr, E. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+effects+of+calvarial+damage+on+dural+mast+cells,+pial+vascular+permeability,+and+cerebral+cortical+histamine+levels+in+rats+and+mice.">Acute effects of calvarial damage on dural mast cells, pial vascular permeability, and cerebral cortical histamine levels in rats and mice.</a> Journal of Neurotrauma. 25 (1), 52-61 (2008).</li><li>Theoharides, T. C., Donelan, J., Kandere-Grzybowska, K., Konstantinidou, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+mast+cells+in+migraine+pathophysiology.">The role of mast cells in migraine pathophysiology.</a> Brain Research Reviews. 49 (1), 65-76 (2005).</li><li>Conti, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Progression+in+migraine:+Role+of+mast+cells+and+pro-inflammatory+and+anti-inflammatory+cytokines.">Progression in migraine: Role of mast cells and pro-inflammatory and anti-inflammatory cytokines.</a> European Journal of Pharmacology. 844, 87-94 (2019).</li><li>Rea, B. 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尽管缺乏组织损伤，但硬脑膜中局部伤害感受系统的适应不良变化被认为是偏头痛发作头痛阶段的关键因素25，26。在这里，该研究提出了一种方法，其中对硬脑膜的微创刺激可以诱导面部触觉过敏。阐明硬膜伤害感受器激活所涉及的机制和事件而不对颅骨和组织造成损害，可以更准确地反映临床前模型中的偏头痛机制。
开颅术和?...

偏头痛仍然是世界范围内的一个主要公共卫生问题。世界卫生组织将其列为世界上第六大流行疾病，折磨着地球人口的不到15%1 ，并对社会造成巨大的社会经济负担2，3。治疗方案及其疗效并不理想，只能缓解症状，不会显著改变偏头痛发生率以下的病理生理学事件4，5。缺乏治疗成功可能是由于偏头痛是一种多因素疾病，其病理学知之甚少，导致治疗靶点数量有限。在动物模型中完全捕获偏头痛也具有挑战性，特别是考虑到偏头痛诊断是基于与患者进行口头交流，这些患者描述了他们对偏头痛特征（如先兆，头痛，畏光和异常疼痛）的经历。尽管如此，重要的是要注意，偏头痛治疗的最新进展目前优于许多神经系统疾病的治疗，这些疾病已被临床前模型充分验证。例如，靶向降钙素基因相关肽或其受体的单克隆抗体和小分子在改善偏头痛患者的生活质量方面非常成功，并可能改变偏头痛的临床管理。虽然在理解这种疾病方面取得了进展，但仍有许多问题有待阐明。
根据临床前动物模型和人类研究，人们普遍认为偏头痛是由脑膜内伤害性纤维的异常激活引起的，这些纤维通过三叉神经和上颈背根神经节6，7，8，9，10发出信号。尽管有这一理论，许多研究仍然使用全身给药来了解偏头痛的潜在促成机制。虽然药物的全身给药大大加强了我们的理解，但这些发现并没有直接评估目标组织内的局部作用是否在偏头痛中起作用。相反，有几项研究采取了刺激硬脑膜的方法;然而，这些实验需要通过侵入性开颅术植入套管，并将恢复时间延长11，12。由于这些局限性，我们开发了一种微创方法来局部刺激硬脑膜，其中缺乏开颅手术消除了手术后的恢复，并允许在清醒的动物中立即进行测试12，13，14。这些注射在轻度异氟醚麻醉下进行，并在小鼠矢状和兰类缝合线的交界处施用。
已经开发了几种方法来评估啮齿动物的伤害感受行为反应15。据报道，大约80%的偏头痛患者的皮肤异常痛16，17 ，代表了用于啮齿动物的潜在转化终点。在临床前模型中，冯弗雷细丝在啮齿动物爪子足底区域的应用已被用于评估临床前偏头痛模型中的疼痛行为。这种方法的主要局限性是它不测试头区域。面部鬼脸评分已用于通过分析诱导疼痛刺激后的面部表情来捕获啮齿动物的疼痛行为18，19。然而，其局限性包括仅捕获对急性刺激的反应，而不是慢性口面部疼痛状况。面部修饰和减少饲养也被认为是偏头痛临床前模型中行为反应的输出20，21。前者的局限性包括难以将疼痛反应与正常的常规修饰和其他感觉（如瘙痒）区分开来。在后者的情况下，在将啮齿动物引入新环境后，饲养行为通常会迅速下降。虽然这些行为终点中的每一个在理解导致疼痛状况的各种机制方面都很有价值，但迫切需要偏头痛等疼痛疾病的临床前模型包括专门捕获头部超敏反应的终点。评估硬膜刺激后眶周皮肤的触觉超敏反应是一种可以更好地了解导致偏头痛的机制的方法，其中感觉症状主要是头部性。在这里，我们描述了一种将物质施用到小鼠硬脑膜上的方法，作为偏头痛的临床前模型。在硬膜应用之后，我们还提出了一种详细的方法，用于测试眶周触觉超敏反应，使用Dixon上下法中应用的校准von Frey细丝。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>4 oz Hot Paper Cups</td>
			<td>Choice Paper Company</td>
			<td>5004W</td>
			<td>https://www.webstaurantstore.com/choice-4-oz-white-poly-paper-hot-cup-case/5004W.html</td>
		</tr>
		<tr>
			<td>Absorbent Underpads</td>
			<td>Fisherbrand</td>
			<td>14-206-65</td>
			<td>https://www.fishersci.com/shop/products/fisherbrand-absorbent-underpads-8/p-306048</td>
		</tr>
		<tr>
			<td>C313I/SPC Internal 28 G cannula</td>
			<td>P1 Technologies (formerly Plastics One)</td>
			<td>8IC313ISPCXC</td>
			<td>I.D. 18 mm, O.D. 35 mm</td>
		</tr>
		<tr>
			<td>Gastight Model 1701 SN Syringes</td>
			<td>Hamilton</td>
			<td>80008</td>
			<td>https://www.hamiltoncompany.com/laboratory-products/syringes/80008</td>
		</tr>
		<tr>
			<td>Ismatec Pump Tubing, 0.19 mm</td>
			<td>Cole-Palmer</td>
			<td>EW-96460-10</td>
			<td>https://www.coleparmer.com/i/ismatec-pump-tubing-2-stop-tygon-s3-e-lab-0-19-mm-id-12-pk/9646010</td>
		</tr>
		<tr>
			<td>Stand with chicken wire</td>
			<td>Custom</td>
			<td></td>
			<td>The galvanized steel chicken wire dimensions are 0.25 in. x 19-gauge</td>
		</tr>
		<tr>
			<td>Testing Rack with individual&#160; Chambers</td>
			<td>Custom</td>
			<td></td>
			<td>Each chamber should have a division between each mouse and lids to contain the mouse. The chambers should also be large enough to hold a 4 oz. paper cup.</td>
		</tr>
		<tr>
			<td>von Frey Filaments</td>
			<td>Touch test/Stoelting</td>
			<td>58011</td>
			<td>https://www.stoeltingco.com/touch-test.html</td>
		</tr>
	</tbody>
</table>

dural stimulation and periorbital von frey testing in mice as a preclinical model of headache

颅脑膜由硬脑膜、蛛网膜和软脑膜组成，被认为主要服务于神经系统的结构功能。 例如，它们保护大脑免受颅骨的侵害，并锚定/组织皮层的血管和神经元供应。然而，脑膜也与偏头痛等神经系统疾病有关，偏头痛期间经历的疼痛归因于局部无菌炎症和随后局部伤害感受传入物的激活。在脑膜的各层中，硬脑膜在偏头痛的病理生理学中特别重要。它是高度血管化的，含有局部伤害感受神经元，并且是各种常驻细胞（如免疫细胞）的家园。局部脑膜微环境的细微变化可能导致硬膜血管周围伤害感受器的激活和致敏，从而导致偏头痛。研究试图通过使用体内电生理学，成像技术或行为模型来解决硬脑膜传入物如何被激活/致敏，但这些通常需要非常侵入性的手术。该协议提出了一种在小鼠硬脑膜上相对非侵入性地应用化合物的方法，以及一种在硬膜刺激后使用眶周von Frey测试测量头痛样触觉敏感性的合适方法。该方法保持硬脑膜和颅骨的完整性，并通过未融合的矢状和兰类缝合线交界处的0.65mm改性套管注射物质，从而减少侵入性技术的混杂效应。这种临床前模型将使研究人员能够研究广泛的硬膜刺激及其在偏头痛病理进展中的作用，例如伤害感受器激活，免疫细胞激活，血管变化和疼痛行为，同时保持颅骨和脑膜的无损伤条件。

这种注射方法用于在小鼠的硬脑膜上施用刺激，以便可能发生随后的行为测试。用这个模型测量的最常见的行为输出是通过von Frey 12，13，14评估的皮肤面部超敏反应。在这里，我们展示了如何使用该模型来评估偏头痛病理学的潜在性别特异性贡献（图3）。
该手术已用于检查硬膜催乳...

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Dural Stimulation and Periorbital von Frey Testing in Mice As a Preclinical Model of Headache

偏头痛最显着的症状是严重的头部疼痛，据推测这是由支配脑膜的感觉神经元介导的。在这里，我们提出了一种以微创方式局部将物质应用于硬脑膜的方法，同时使用面部超敏反应作为输出。

小鼠硬脑膜刺激和眶周冯弗雷测试作为头痛的临床前模型

The cranial meninges, comprised of the dura mater, arachnoid, and pia mater, are thought to primarily serve structural functions for the nervous system.  ...

在没有组织损伤的情况下，脑膜内发生哪些事件导致头痛？与先前在大鼠中使用硬脑膜刺激模型不同，它允许使用转基因小鼠的广泛目录。主要的斗争是通过用注射器触诊颅骨来定位兰氏体和矢状缝合线的交点。首先，通过修改市售的内部套管和带有熔融石英塑料盖的注入器来创建小鼠注入器，该盖子可调节并插入28号导轨套管中。使用卡钳调整熔融石英塑料盖在注入器上，而无需弯曲注入器。将注入器调整为从注入器尖端到硅塑料盖边缘测量的6毫米长度。对其他小鼠品系进行先导注射，并调整长度以精确注射硬脑膜。然后，将注入器的长端连接到八英寸或更大的塑料管上，以容纳五微升的体积。插入管子，同时确保完全覆盖金属部分和位于注入器上的塑料塞子的顶部，以防止气泡积聚在管路中。接下来，填充微量注射器并将管子的另一端连接到10微升玻璃微量注射器上，确保注射器的金属部分紧密密封。然后，用五微升PBS或合成间质液或其他选择的载体回灌注射器，以防止气泡形成。如果需要，用车辆淹没线路以消散预成型气泡。或者，预先填充注射器，连接后，将流体推过连接的管路。一旦车辆填充的生产线高效工作，将五微升溶液装入汉密尔顿注射器。一旦小鼠完全麻醉，将无菌眼药膏涂抹在眼睛上并剃掉动物的头部，然后用聚维酮碘和乙醇消毒皮肤。用一只手稳定动物的头部，另一只手用装有药物的注射器握住注入器。然后，使用注入器，仔细探测，定位并验证小鼠颅骨上矢状缝合线和兰氏缝合线的交界处。一旦找到缝合线并且注入器就位，缓慢而轻轻地来回摆动注入器，直到注入器穿过皮肤并下降到连接处，一直到塑料塞子，确保将整个6毫米注入器尖端插入连接处。然后，慢慢推动注射器的柱塞，将五微升液体排出到硬脑膜中。在测试当天，适应房间后，从相应的腔室中取出一只鼠标和杯子。将杯子保持在水平位置，使体重均匀分布，因为不相等的重量分布可能会改变或阻止动物的反应。将装有鼠标的杯子放在测试架下方桌子上的吸收垫上。然后，对于眶周冯弗雷测试，将07克冯弗雷丝放在眼睛之间的面部中心，并对细丝施加足够的压力，使冯弗雷头发弯曲成C形。保持细丝接触约三到五秒钟，或直到鼠标使用头部或爪子滑动掉细丝。要遵循Dixon上下方法，如果鼠标对灯丝有反应，请应用下一个重量较低的细丝，否则，应用重量较高的细丝。该手术用于检查硬膜催乳素对机械诱发的面部超敏反应的影响。该研究的结果表明，雌性ICR小鼠对五微克硬膜催乳素的反应显着降低了面部戒断阈值。低十倍剂量的5微克催乳素也显示出面部戒断阈值降低，类似于高剂量催乳素。催乳素注射产生自发性疼痛相关行为，通过鬼脸进行评估。硬脑膜5微克催乳素在雌性小鼠中引起显着的鬼脸，并进一步证明了硬膜催乳素在雌性偏头痛样行为中的明确作用 行为测试使用von Frey丝或面部鬼脸来确定刺激在没有损伤的情况下是否引起头痛反应。该技术现在可用于研究脑膜中基于炎症的机制，而不用担心由颅骨切开术和随后的炎症引起的警告。

Research

JoVE Journal

Neuroscience

Combining Transcranial Magnetic Stimulation and fMRI to Examine the Default Mode Network

结合经颅磁刺激和功能磁共振成像检查默认模式网络

The default mode network is a group of brain regions that are active when an individual is not focused on the outside world and the brain is at "wakeful ...

科研

神经科学

TMS: Using the Theta-Burst Protocol to Explore Mechanism of Plasticity in Individuals with Fragile X Syndrome and Autism

TMS：使用的Theta突发协议，以探索个人与脆性X综合征与孤独症的可塑性Mechasnism

Fragile X Syndrome (FXS), also known as Martin-Bell Syndrome, is a genetic abnormality found on the X chromosome.1,2 Individuals suffering from FXS ...

The NeuroStar TMS Device: Conducting the FDA Approved Protocol for Treatment of Depression

NeuroStar TMS的设备进行美国FDA批准用于治疗抑郁症的议定书“

The Neuronetics NeuroStar Transcranial Magnetic Stimulation (TMS) System is a class II medical device that produces brief duration, pulsed magnetic ...

Electrode Positioning and Montage in Transcranial Direct Current Stimulation

经颅直接电流刺激电极定位和蒙太奇

Transcranial direct current stimulation (tDCS) is a technique that has been intensively investigated in the past decade as this method offers a ...

Measuring Changes in Tactile Sensitivity in the Hind Paw of Mice Using an Electronic von Frey Apparatus

Measuring inflammation-induced changes in thresholds of hind paw withdrawal from mechanical pressure is a useful technique to assess changes in pain ...

The Olfactory System as a Model to Study Axonal Growth Patterns and Morphology In Vivo

The Olfactory System as a Model to Study Axonal Growth Patterns and Morphology In Vivo

嗅觉系统作为模型来研究轴突生长的模式和形态<em&gt;在体内</em

The olfactory system has the unusual capacity to generate new neurons throughout the lifetime of an organism. Olfactory stem cells in the basal portion of ...

A Simple and Inexpensive Method for Determining Cold Sensitivity and Adaptation in Mice

测定冷灵敏度和适应小鼠一种简单廉价的方法

Cold hypersensitivity is a serious clinical problem, affecting a broad subset of patients and causing significant decreases in quality of life.  The cold ...

Simultaneous ex vivo Functional Testing of Two Retinas by in vivo Electroretinogram System

Simultaneous ex vivo Functional Testing of Two Retinas by in vivo Electroretinogram System

同时<em&gt;体外</em两个由视网膜&gt;功能测试<em&gt;在体内</em&gt;电图系统

An In vivo electroretinogram (ERG) signal is composed of several overlapping components originating from different retinal cell types, as well as noise ...

An In Ovo Model for Testing Insulin-mimetic Compounds

An In Ovo Model for Testing Insulin-mimetic Compounds

用于测试胰岛素模拟化合物的蛋模型

Elevated blood glucose levels in type 2 diabetes mellitus (T2DM), a complex and multifactorial metabolic disease, are caused by insulin resistance and ...

Transcranial Direct Current Stimulation (tDCS) in Mice

Transcranial Direct Current Stimulation tDCS in Mice

经颅直流电刺激 (tDCS) 在小鼠中的作用

Transcranial direct current stimulation (tDCS) is a non-invasive neuromodulation technique proposed as an alternative or complementary treatment for ...