Name: measurement of tactile allodynia in a murine model of bacterial prostatitis
Uploaded: 2013-01-16T13:00:00.000+00:00
Duration: 6 min 46 s
Description: Watch this Scientific Journal Video about Measurement of Tactile Allodynia in a Murine Model of Bacterial Prostatitis at JoVE.com

这项研究是由美国国立卫生研究院/ NIDDK由授予1K01DK079019A2（PT）。

1。小鼠接种细菌的准备工作以下，必须在无菌条件下完成。 <ol><li>拿一个17×100毫米聚丙烯管帽和加入3毫升的新鲜Luria培养基（LB）媒体。使用蒸压提示，采取一些冷冻的细菌甘油CP1 2株，少量的文化转移到管中的LB培养基。更换的管帽，这样的氧气仍然可以进入管 - 文化需要在有氧条件下生长。将管在37℃下摇床中在220rpm过夜。 </li><li>第二天，5微升过夜培养物转移到新的管中，用3毫升的LB培养基，并在37℃的培养箱中培养过夜的静态条件下成长</li><li>在第3天转移到AA 50ml管40μl的文化含40毫升LB媒体。在37°C的静态培养箱中培养过夜地点。 </li><li>打开并设置离心机至4℃。一旦离心机已达到最终温度，每40毫升的文化转移到40毫升的Nalgene离心管中。 </li><li>称取离心管，以平衡 - 调节音量LB媒体。在6,000 rpm，20分钟的自旋管。 </li><li>仔细收集上清，轻轻地暂停细菌沉淀在40毫升无菌PBS。 </li><li>重复步骤1.5 PBS。 </li><li>吸出上清液，并暂停细菌在500μl无菌PBS。将该溶液转移到1.5毫升微量离心管中。取10微升的悬浮液，并在990微升冰冷的PBS中稀释。使用分光光度计读取OD 420 nm处的体积，以确定所需的接种。要确定适当体积的冰冷的PBS中止颗粒：OD 420 nm的数量（毫升）和减去估计量的细菌沉淀前。 OD 420 nm处 = 0.545，颗粒约0.075毫升。 0.545 - 0.075 = 0.470]。 </li><li>取出10μl的细菌悬浮液和二琵琶在990微升冰冷的PBS。外径420纳米 。我们的目标是实现稀释后的悬浮液中1.000±0.010的OD 420 nm处的值。如果您的稀释的数量超过这个目标，增加更多的体积悬浮液中，采取另一种解读。如果数目是低于0.990然后降速的悬浮液，并重复OD 420 nm处读数，直到获得所需的读数。 </li></ol> 2。动物感染小鼠（5〜7周龄，10％组）均购自巴港的杰克逊实验室（ME）。 <ol><li>小鼠麻醉吸入1-4％异氟醚一个有机玻璃室中，被监视，直到卧位。 </li><li>在一个时间的一个鼠标滴注细菌采取导尿用聚乙烯（PE10）的导管连接到的变形30 G针的玻璃Hamilton注射器（长度为1.5-2.0厘米）。导管被插入到轮毂的针。鼠标是放置其背表面麻醉下用鼻锥保持。 </li><li>用于润滑的入口到阴茎尿道，阴茎的鼠标是温和的压力和自由量的润滑剂棉签挤出。 </li><li> 10μl的磷酸盐缓冲生理盐水中含1×10 8个细菌引入到尿道后麻醉小鼠导尿。 </li><li>维持小鼠在麻醉的状态下的细菌引进的有机玻璃室后15分钟，让细菌附着，防止即时排尿。 </li><li>小鼠被放置在笼子里，并在接下来的24小时监控。 </li></ol> 3。行为测试小鼠在感染前（基线）和感染后3，7，14，21和28进行了测试。简称痛觉过敏和触觉异常进行了测试使用von Frey细丝适用于腹部11,12和足底ŕ对旅游区的后掌13。测试perfomed在一个固定的时间的，标准的方法和单实验者测试的所有动物。盲法测试组被用来打击在动物模型的局限性，基于行为的疼痛测试。五个单独的von Frey细丝与力量，0.04，0.16，0.4，1.0和4.0克（Stoelting，美国）被施加到撤回的腹部和频率响应计算。 <ol><li>的驯化后，30分钟内，被放置在个别有机玻璃腔室（6×10×12厘米），在内部用不锈钢线栅地板小鼠。 </li><li>采用von Frey细丝简称痛觉过敏和触觉异常。各灯丝施加为1-2秒的总共有10倍刺激间的间隔为5秒，武力升序顺序和毛发进行了测试。 </li><li>每个灯丝被施加到下腹部区域中的一般的前列腺附近护理刺激不同的区域，在此区域内，以避免的脱敏或“风”的影响。适用于力1-2秒递增的顺序与刺激之间的间隔至少为5秒的10倍，总的细丝。 </li><li>三种类型的行为被认为是一个积极的回应长丝刺激：1）急剧收缩腹部; 2）直接舔或，搔抓的长丝刺激; 3）跳跃。 </li><li>肯定响应（满分10， 例如 ，5的反应10 = 50％）的百分比计算，并响应频率数据被报告为响应频率的平均百分比±SE </li><li>超过25个阳性总基线反应的动物被排除在研究之外。 </li><li>用von Frey细丝的后肢力量，0.04，0.16，0.4，1.0和4.0克的的足底地区的触觉异常性疼痛进行了测试。中位数的50％收回阈值（5）进行了评估使用的上下方法在测试开始0.04克长丝的后掌跖面垂直，直到灯丝稍微弯曲。升序排列，直到一个积极的反应，观察单丝测试。积极应对的灯丝被定义为测试爪爪或舔的大幅撤出。当一个正响应记录下一个较弱施加长丝，如果观察到的否定响应，则下一个较强的长丝。 </li><li>的实验和描述的方法进行了审查，并批准由西北大学动物照顾及使用委员会。 </li></ol>

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感染的小鼠前列腺UPEC允许为在体内建模可能涉及在细菌性前列腺炎的发病机制中，CP / CPPS或作为慢性炎症的诱发事件的事件。 UPEC模型描述的方法对细菌制剂和滴注战平后，庞大的身躯文学中女性尿路感染7,14。该模型具有广泛的适用性研究发病机制，检测潜在的候选疫苗和免疫调节机制。疼痛行为的能力，按照可量化的方式使感染引起的疼痛发病机制的研究，并有可能作为测试疼痛治疗的临床前模型。 有几个关键的步骤，确定成功的小鼠感染模型。振荡和静态培养为期三天的培养方法进行菌毛的最佳表达，是已知的是重要的UPEC连接到上皮细胞。它代表了一种成功的器官定植的关键步骤，统一企业。细菌生长的技术已被专门设计UPEC生长与优化的OD 420的步骤用于获得10μl的磷酸缓冲盐水中含1×10 8个细菌7。其他种或菌株的细菌可能以不同的速率生长和标准化将是重要的导出适当的外径，使1×10 8个细菌。种不同的细菌坚持以独特的方式在很大程度上依赖于该菌株的毒力因子表达14在膀胱和前列腺的细胞。因此，成功感染的小鼠前列腺是依赖于利用UPEC菌株的毒力因子与适当的补充。紧随感染，至少15分钟，保持在轻度麻醉状态下的动物是非常重要的，以确保插件耕种的细菌有时间附加，并开始发病前的正常机制的注入量从尿道排尿清除。 在小鼠体内的触觉异常是一个结果的基础内脏病理的特点是精致的宿主菌的特异性。我们前面描述的UPEC，在雄性NOD / ShiLtJ的小鼠的致病性菌株引起异常性疼痛3天达到峰值，并维持长期而一脉相承的是不能诱导异常性疼痛在C57BL/67小鼠2。我们的研究表明自身免疫过程中的NOD / ShiLtJ小鼠的根本原因的UPEC引起的加速。因此，宿主的遗传背景的触觉异常的发展是很重要的。其他重要的步骤是公正和测量成功的痛觉超敏的关键是确保测试仪的身份蒙蔽的治疗，相同的测试人员是负责测量的所有时间-Points的实验，利用类似的测试时间在白天，住房条件的小鼠并确保时间内点的标准条件下，在一个实验中，以及实验之间。 感染小鼠模型有一个数量的限制，在结果的解释需要慎重考虑的。灌输的细菌感染以及膀胱前列腺尿道内有能力。这种复杂的解释的基本病理任何全身或一般的参数，可以从多个器官。然而临床和前列腺癌来源的细菌菌株，利用以及允许并发检查的膀胱和前列腺的适当的解释。此外，感染的方法，模拟人类的男性可能上行感染方式。 感染模型不同于先前报道PRI玛利莲株UPEC使用，主机的小鼠品系和不同的文化技术和体积的细菌灌输到尿道15。本研究采用了良好的特点源性前列腺慢性前列腺炎UPEC菌株进行调解疾病。2。早先的研究已经使用了细菌来自膀胱感染或急性前列腺炎主要是为了利用研究炎症相关的事件，但在的触觉异常6,15的没有特点。许多其他的动物模型已经报道，利用自身免疫机制5,16，激素处理17和新生儿thymectomies 18日至诱发前列腺炎。虽然这些模型有一定的积极属性，包括器官特异性疾病的病理和利用现有的技术，这些方法的实际发病的CP / CPPS的相关性是未知的。相反，所描述的方法在这个人uscript是指已被假定是在前列腺疾病的发病机制的引发剂的一个潜在的机制。此外，超出了它的效用理解CP / CPPS发病，技术很可能被用来在建立和检查对前列腺的慢性炎症，良性前列腺增生症（BPH）和前列腺癌中的作用。

<table border="1"><tbody><tr><td> 的试剂的名称 </td><td> 公司 </td><td> 目录编号 </td></tr><tr><td>文化管</td><td> BD猎鹰</td><td> 352059 </td></tr><tr><td> LB肉汤米勒</td><td> EMD </td><td> EM1.10285.0500 </td></tr><tr><td> NALGENE离心管</td><td>热</td><td> 3118-0050 </td></tr><tr><td>异氟醚</td><td>巴特勒施恩</td><td> NDC 11695-6776-1 </td></tr><tr><td>导管针30G </td><td>汉密尔顿</td><td> 91030 </td></tr><tr><td> PE10管材</td><td> BD intramedic </td><td> 427400 </td></tr><tr><td> von Frey细丝</td><td> Stoelting </td><td> 58025-31 </td></tr></tbody></table>

measurement of tactile allodynia in a murine model of bacterial prostatitis

致肾盂肾炎大肠杆菌 （UPEC）是病原体中发挥了重要的作用，尿路感染和细菌性前列腺炎1。最近，我们已经显示，UPEC有重要的作用开始在慢性盆腔疼痛2，功能的慢性前列腺炎/慢性盆腔疼痛综合征（CP / CPPS）3,4。感染前列腺的临床相关UPEC可以发起和建立慢性疼痛的机制可能涉及组织损伤和5的自身免疫机制的启动。 了解的统一企业在前列腺癌的发病机制是一个挑战，相对交通不便的前列腺操作。我们利用先前描述的尿道感染的方法提供临床株UPEC雄性小鼠，从而建立一个上行性感染的前列腺。在这里，我们描述的协议规范BActerial接种7以及滴注细菌置管麻醉的雄性小鼠的程序。 CP / CPPS的主要特征是由触觉异常4的存在下。行为测试是基于皮肤痛觉过敏的概念称为内脏痛8-10。易怒集中在内脏器官，减少皮肤的痛阈值，允许夸张的反应，正常的非疼痛刺激（痛觉超敏）。正常的力中的应用皮肤导致异常反应，往往增加与强度的底层的内脏痛。我们描述在的NOD / ShiLtJ小鼠，利用von弗雷纤维中的量化触觉异常性疼痛的随着时间的推移，在响应于一个单一的UPEC细菌感染的方法。

<html>我们审查NOD / ShiLtj的C57BL/6J雄性小鼠的存在细菌感染后的慢性疼痛。雄性小鼠（5至7周龄），用生理盐水或细菌进入尿道（ 图1），灌输。机械性刺激导致的骨盆区，用von Frey细丝的生理盐水注入小鼠和UPEC感染的小鼠在28天的试验过程中（ 图1）的响应频率并没有改变。 UPEC感染的NOD小鼠表现出与此相反，是显着更大的盆腔刺激的反应仍然显着升高，直到第28天（P &lt;0.05， 图3）。细菌感染导致触觉后肢足底区域（数据未示出）没有变化。 tp_upload/50158/50158fig1.jpg“/&gt; 图1。实验流程图。1×10 8个细菌悬浮在为期三天的文化制备PBS，并逐步进入尿道口的麻醉小鼠导尿。使用von Frey细丝与力量，0.04，0.16，0.4，1，和4克所得触觉异常进行定量。 <img alt="图2" src="/files/ftp_upload/50158/50158fig2.jpg" /> 图2。测试方法论用于测试触觉异常使用von Frey纤维。小鼠的细菌感染前和感染后日（PIDS），7，14，21，和28。三种不同类型的行为被认为是作为灯丝刺激的积极回应：1）急剧收缩腹部，2）直接舔或划伤的面积长丝刺激;或3）跳跃。响应经常被计算的百分比的积极响应（满分10分）和DATA报告的响应频率±SE的平均百分比。 <img alt="图3" src="/files/ftp_upload/50158/50158fig3.jpg" /> 图3。触觉异常性疼痛引起的的UPEC细菌。提到内脏痛觉过敏的骨盆区域使用von Frey细丝的5个校准力量对机械刺激的反应。数据的平均百分比的积极响应±SE之前灌注的细菌（基线）和的PID 7日，14日，21日和28日在报道。方差分析表示响应频率的显着增加，在7至28的PID，与在测试中UPEC感染的NOD小鼠的（P &lt;0.05）的所有长丝的基线相比。在每个PID％的应答计算为所有纤维相对于基线反应的总反应。

Watch this Scientific Journal Video about Measurement of Tactile Allodynia in a Murine Model of Bacterial Prostatitis at JoVE.com

Measurement of Tactile Allodynia in a Murine Model of Bacterial Prostatitis

在调解盆腔疼痛慢性前列腺炎前列腺感染可能是一个促进因素。我们的程序编制标准化的细菌接种的细菌进入尿道的雄性小鼠和方法测量触觉异常的小鼠，随着时间的推移，滴入。

细菌性前列腺炎小鼠模型的测量触觉异常

Uropathogenic Escherichia coli (UPEC) are pathogens that play an important role in urinary tract infections and bacterial prostatitis1. We have recently ...

measurement-tactile-allodynia-murine-model-bacterial

Universidad San Sebastian

Research

JoVE Journal

Immunology and Infection

15.8K 次观看.  Northwestern University Feinberg School of Medicine. 在调解盆腔疼痛慢性前列腺炎前列腺感染可能是一个促进因素。我们的程序编制标准化的细菌接种的细菌进入尿道的雄性小鼠和方法测量触觉异常的小鼠，随着时间的推移，滴入。

视频: Measurement of Tactile Allodynia in a Murine Model of Bacterial Prostatitis

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 ...

科研

行为学

The Sciatic Nerve Cuffing Model of Neuropathic Pain in Mice

Neuropathic pain arises as a consequence of a lesion or a disease affecting the somatosensory system. This syndrome results from maladaptive changes in injured sensory neurons and along the entire nociceptive pathway within the central nervous system. It is usually chronic and challenging to treat. In order to study neuropathic pain and its treatments, different models have been developed in rodents. These models derive from known etiologies, thus reproducing peripheral nerve injuries, central injuries, and metabolic-, infectious- or chemotherapy-related neuropathies. Murine models of peripheral nerve injury often target the sciatic nerve which is easy to access and allows nociceptive tests on the hind paw. These models rely on a compression and/or a section. Here, the detailed surgery procedure for the &#34;cuff model&#34; of neuropathic pain in mice is described. In this model, a cuff of PE-20 polyethylene tubing of standardized length (2 mm) is unilaterally implanted around the main branch of the sciatic nerve. It induces a long-lasting mechanical allodynia, i.e., a nociceptive response to a normally non-nociceptive stimulus that can be evaluated by using von Frey filaments. Besides the detailed surgery and testing procedures, the interest of this model for the study of neuropathic pain mechanism, for the study of neuropathic pain sensory and anxiodepressive aspects, and for the study of neuropathic pain treatments are also discussed.

Neuropathic pain is a consequence of a lesion or disease affecting the somatosensory system. The &#8220;cuff model&#8221; of neuropathic pain in mice consists of the implantation of a polyethylene cuff around the main branch of the sciatic nerve. Mechanical allodynia is tested using von Frey filaments.

神经病理性疼痛小鼠坐骨神经掴型号

Neuropathic pain arises as a consequence of a lesion or a disease affecting the somatosensory system. This syndrome results from maladaptive changes in ...

医学

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 perception in rodents. Withdrawal thresholds can be measured first at baseline and then following drug, venom, injury, allergen,&#160;or otherwise evoked inflammation by applying an accurate force on very specific areas of the skin. An electronic von Frey apparatus allows precise assessment of mouse hind paw withdrawal thresholds that are not limited by the available filament sizes in contrast to classical von Frey measurements. The ease and rapidity of measurements allow for incorporation of assessment of tactile sensitivity outcomes in diverse models of rapid-onset inflammatory and neuropathic pain as multiple measurements can be taken within a short time period. Experimental measurements for individual rodent subjects can be internally controlled against individual baseline responses and exclusion criteria easily established to standardize baseline responses within and across experimental groups. Thus, measurements using an electronic von Frey apparatus represent a useful modification of the well-established classical von Frey filament-based assays for rodent mechanical allodynia that may also be applied to other nonhuman mammalian models.

Electronic von Frey measurements are an effective and improved alternative to classical von Frey filaments, as they allow rapid and precise measurement of changes in rodent mechanical withdrawal thresholds to continuously applied pressure stimuli before and after an inflammatory event.

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

神经科学

An Improved Assay and Tools for Measuring Mechanical Nociception in Drosophila Larvae

Published assays for mechanical nociception in Drosophila have led to variable assessments of behavior. Here, we fabricated, for use with Drosophila larvae, customized metal nickel-titanium alloy (nitinol) filaments. These mechanical probes are similar to the von Frey filaments used in vertebrates to measure mechanical nociception. Here, we demonstrate how to make and calibrate these mechanical probes and how to generate a full behavioral dose-response from subthreshold (innocuous or non-noxious range) to suprathreshold (low to high noxious range) stimuli. To demonstrate the utility of the probes, we investigated tissue damage-induced hypersensitivity in Drosophila larvae. Mechanical allodynia (hypersensitivity to a normally innocuous mechanical stimulus) and hyperalgesia (exaggerated responsiveness to a noxious mechanical stimulus) have not yet been established in Drosophila larvae. Using mechanical probes that are normally innocuous or probes that typically elicit an aversive behavior, we found that Drosophila larvae develop mechanical hypersensitization (both allodynia and hyperalgesia) after tissue damage. Thus, the mechanical probes and assay that we illustrate here will likely be important tools to dissect the fundamental molecular/genetic mechanisms of mechanical hypersensitivity.

An Improved Assay and Tools for Measuring Mechanical Nociception in Drosophila Larvae

The goal of this protocol is to show how to perform an improved assay for mechanical nociception in Drosophila larvae. We use the assay here to demonstrate that mechanical hypersensitivity (allodynia and hyperalgesia) exists in Drosophila larvae.

测量 果蝇 拉瓦的机械结节的改进的检测和工具

Published assays for mechanical nociception in Drosophila have led to variable assessments of behavior. Here, we fabricated, for use with Drosophila ...

Assessment of Morphine-induced Hyperalgesia and Analgesic Tolerance in Mice Using Thermal and Mechanical Nociceptive Modalities

Opioid-induced hyperalgesia and tolerance severely impact the clinical efficacy of opiates as pain relievers in animals and humans. The molecular mechanisms underlying both phenomena are not well understood and their elucidation should benefit from the study of animal models and from the design of appropriate experimental protocols. 
We describe here a methodological approach for inducing, recording and quantifying morphine-induced hyperalgesia as well as for evidencing analgesic tolerance, using the tail-immersion and tail pressure tests in wild-type mice. As shown in the video, the protocol is divided into five sequential steps. Handling and habituation phases allow a safe determination of the basal nociceptive response of the animals. Chronic morphine administration induces significant hyperalgesia as shown by an increase in both thermal and mechanical sensitivity, whereas the comparison of analgesia time-courses after acute or repeated morphine treatment clearly indicates the development of tolerance manifested by a decline in analgesic response amplitude. This protocol may be similarly adapted to genetically modified mice in order to evaluate the role of individual genes in the modulation of nociception and morphine analgesia. It also provides a model system to investigate the effectiveness of potential therapeutic agents to improve opiate analgesic efficacy.

We describe a protocol to examine the development of opioid-induced hyperalgesia and tolerance in mice. Based on the measurement of thermal and mechanical nociceptive responses of na&#239;ve and morphine-treated animals, it allows to quantify the increase in pain sensitivity (hyperalgesia) and decrease in analgesia (tolerance) associated with chronic opiate administration.

吗啡诱发的痛觉过敏和镇痛耐受小鼠中利用热和机械伤害性的评估方式

Opioid-induced hyperalgesia and tolerance severely impact the clinical efficacy of opiates as pain relievers in animals and humans. The molecular ...

Back Mechanical Sensitivity Assessment in the Rat for Mechanistic Investigation of Chronic Back Pain

Low back pain is the leading cause of disability worldwide, with dramatic personal, economic, and social consequences. To develop novel therapeutics, animal models are needed to examine the mechanisms and effectiveness of novel therapies from a translational perspective. Several rodent models of back pain are used in current investigations. Surprisingly, however, no standardized behavioral test was validated to assess mechanical sensitivity in back pain models. This is critical to confirm that animals with presumed back pain present local hypersensitivity to nociceptive stimuli, and to monitor sensitivity during interventions designed to relieve back pain. The objective of this study is to lay down a simple and accessible test to assess mechanical sensitivity in the back of rats. A test cage was fabricated specifically for this method; length x width x height: 50 x 20 x 7 cm, having a stainless-steel mesh on the top. This test cage allows the application of mechanical stimuli to the back. To perform the test, the back of the animal is shaved in the region of interest, and the test area is marked to repeat the test on different days, as needed. The mechanical threshold is determined with Von Frey filaments applied to the paraspinal muscles, utilizing the up-down method described previously. The positive responses include (1) muscle twitching, (2) arching (back extension), (3) rotation of the neck (4) scratching or licking the back, and (5) escaping. This behavioral test (Back Mechanical Sensitivity (BMS) test) is useful for mechanistic research with rodent models of back pain for the development of therapeutic interventions for the prevention and management of back pain.

To develop novel therapeutic interventions for the prevention and management of back pain, animal models are required to examine the mechanisms and effectiveness of these therapies from a translational perspective. The present protocol describes the BMS test, a standardized method to assess back mechanical sensitivity in the rat.

大鼠背部机械敏感性评估用于慢性背痛的机制研究

Low back pain is the leading cause of disability worldwide, with dramatic personal, economic, and social consequences. To develop novel therapeutics, ...

Urokinase-type Plasminogen Activator-induced Mouse Back Pain Model

A model of persisting lower back pain can be induced in mice with the simple methodology described herein. Step-by-step methods for simple, rapid induction of a persisting back pain model in mice are provided here using an injection of urokinase-type plasminogen activator (urokinase), a serine protease present in humans and other animals. The methodology for induction of persisting lower back pain in mice involves a simple injection of urokinase along the ligamentous insertion region of the lumbar spine. The urokinase inflammatory agent activates plasminogen to plasmin. Typically, the model can be induced within 10 min and hypersensitivity persists for at least 8 weeks.
Hypersensitivity, gait disturbance, and other standard anxiety- and depression-like measures can be tested in the persisting model.&#160;Back pain is the most prevalent type of pain. To improve awareness of back pain, the International Association for the Study of Pain (IASP) named 2021 the &#34;Global Year about Back Pain&#34; and 2022 the &#34;Global Year for Translating Pain Knowledge to Practice.&#34; One limitation of the therapeutic advancement of pain therapeutics is the lack of suitable models for testing persistent and chronic pain. The&#160;features of this model&#160;are suitable for testing potential therapeutics aimed at the reduction of back pain and its ancillary characteristics, contributing to IASP&#39;s naming 2022 as the Global Year for Translating Pain Knowledge to Practice.

Methods for simple, rapid induction of a back pain model in mice are provided here using an intraligament injection of urinary plasminogen activator.

尿激酶型纤溶酶原激活剂诱导的小鼠背痛模型

A model of persisting lower back pain can be induced in mice with the simple methodology described herein. Step-by-step methods for simple, rapid ...

Assessment of Perigenital Sensitivity and Prostatic Mast Cell Activation in a Mouse Model of Neonatal Maternal Separation

Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) has a lifetime prevalence of 14% and is the most common urological diagnosis for men under the age of 50, yet it is the least understood and studied chronic pelvic pain disorder. A significant subset of patients with chronic pelvic pain report having experienced early life stress or abuse, which can markedly affect the functioning and regulation of the hypothalamic-pituitary-adrenal (HPA) axis. Mast cell activation, which has been shown to be increased in both urine and expressed prostatic secretions of CP/CPPS patients, is partially regulated by downstream activation of the HPA axis. Neonatal maternal separation (NMS) has been used for over two decades to study the outcomes of early life stress in rodent models, including changes in the HPA axis and visceral sensitivity. Here we provide a detailed protocol for using NMS as a preclinical model of CP/CPPS in male C57BL/6 mice. We describe the methodology for performing NMS, assessing perigenital mechanical allodynia, and histological evidence of mast cell activation. We also provide evidence that early psychological stress can have long-lasting effects on the male urogenital system in mice.

We are measuring perigenital mechanical sensitivity and mast cell activation in the prostate of male C57BL/6 mice that underwent an early life stress paradigm &#8211; neonatal maternal separation, in order to induce a preclinical model of chronic prostatitis/chronic pelvic pain syndrome.

的Perigenital敏感性和前列腺肥大细胞活化新生母婴分离的小鼠模型评估

Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) has a lifetime prevalence of 14% and is the most common urological diagnosis for men under the ...

Transurethral Instillation Procedure in Adult Male Mouse

Transurethral instillation can be used to deliver different solutions with active ingredients (e.g., drugs, chemicals, bacteria, and viruses) locally into the urinary bladder to either induce animal models of bladder pathologies or evaluate the effectiveness of intravesical treatments. Most rodent models of lower urinary tract (LUT) pathologies are induced in female mice due to ease of intravesical instillation of the substances via the female urethra. However, due to anatomical differences between the female and male LUT, transurethral instillation in a male mouse has been deemed a very challenging procedure, and it has not been previously described. In this manuscript, we provide a detailed description of how to prepare polyethylene (PE) tubing for subsequent insertion into the urethra of a male mouse. In addition, we discuss the ideal types of PE tubing to be used depending on the desired site of inoculation. Furthermore, we describe point by point how to prepare an animal for a successful transurethral instillation to avoid injury to the urethra and ensure the delivery of the solution to the desired location. The procedure is started by retracting the prepuce and the glans to expose the opening of the urethral meatus. Next, the glans are grasped by blunt non-crushing forceps to stabilize the penis and the PE tubing. The PE tubing is first inserted into the urethral meatus parallel to the animal body, then its angle is adjusted by tilting the catheter to maneuver it to follow the natural curvature of the urethra. This technique can be used to induced murine models of bladder pathologies and/or evaluate the effectiveness of intravesical treatments in male mice.

Transurethral instillation is a challenging procedure and has not been well described in the literature. The aim of this manuscript is to describe a technique for transurethral insertion of a catheter for intravesical delivery of liquids with active substances into the urinary bladder and/or prostate in adult male mice.

成年雄性小鼠经尿道灌注术

Transurethral instillation can be used to deliver different solutions with active ingredients (e.g., drugs, chemicals, bacteria, and viruses) locally into ...

免疫与感染

评估 T. evansi 感染动物的疼痛和异常性疼痛

Electronic von Frey associated with the Mouse Grimace Scale to Assess Allodynia and Pain in Trypanosoma evansi-Infected Mice

Infectious disease pathogenesis is still a complex field to study. The course of several clinical signs, such as allodynia and pain, may be observed in domestic animals. However, the knowledge of their pathways and correct treatment need controlled experiments, many of them using laboratory animals. Measuring changes in mechanical thresholds of the hind paw and viscera is a useful technique to observe changes in pain perception in rodents. Withdrawal response can be measured first in baseline tests, which creates better control of experimental groups. Subsequent tests can be performed after inducing infection and adding drugs to the protocol. The use of an electronic von Frey apparatus associated with the use of a facial scale to observe pain-like changes allows a simple, precise, and consistent assessment to evaluate allodynia and pain in mice. Thus, experiments using the present methodology for Trypanosoma evansi infection represent a useful method to evaluate allodynia and pain in laboratory-infected animals, which can be applied to the conventional treatment for livestock animals.

作者聚焦:评估感染埃文斯锥虫的实验动物的疼痛并促进动物福利

Here, we present a protocol to evaluate nociception in mice infected with Trypanosoma evansi, using the electronic von Frey apparatus as a tool, measuring mechanical thresholds of the hind paw and viscera.

电子 von Frey 与小鼠鬼脸量表相关联，以评估 埃文西锥虫感染小鼠的异常性疼痛和疼痛

Infectious disease pathogenesis is still a complex field to study. The course of several clinical signs, such as allodynia and pain, may be observed in ...

细菌性前列腺炎小鼠模型的测量触觉异常

摘要

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此视频中的章节

大鼠复杂区域疼痛综合征I型慢性缺血后疼痛模型

神经病理性疼痛小鼠坐骨神经掴型号

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

测量果蝇拉瓦的机械结节的改进的检测和工具

吗啡诱发的痛觉过敏和镇痛耐受小鼠中利用热和机械伤害性的评估方式

大鼠背部机械敏感性评估用于慢性背痛的机制研究

尿激酶型纤溶酶原激活剂诱导的小鼠背痛模型

的Perigenital敏感性和前列腺肥大细胞活化新生母婴分离的小鼠模型评估

成年雄性小鼠经尿道灌注术

电子 von Frey 与小鼠鬼脸量表相关联，以评估埃文西锥虫感染小鼠的异常性疼痛和疼痛

细菌性前列腺炎小鼠模型的测量触觉异常

摘要

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此视频中的章节

大鼠复杂区域疼痛综合征I型慢性缺血后疼痛模型

神经病理性疼痛小鼠坐骨神经掴型号

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

测量 果蝇 拉瓦的机械结节的改进的检测和工具

吗啡诱发的痛觉过敏和镇痛耐受小鼠中利用热和机械伤害性的评估方式

大鼠背部机械敏感性评估用于慢性背痛的机制研究

尿激酶型纤溶酶原激活剂诱导的小鼠背痛模型

的Perigenital敏感性和前列腺肥大细胞活化新生母婴分离的小鼠模型评估

成年雄性小鼠经尿道灌注术

电子 von Frey 与小鼠鬼脸量表相关联，以评估 埃文西锥虫感染小鼠的异常性疼痛和疼痛

测量果蝇拉瓦的机械结节的改进的检测和工具

电子 von Frey 与小鼠鬼脸量表相关联，以评估埃文西锥虫感染小鼠的异常性疼痛和疼痛