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

이 연구는 NIH / NIDDK에서 부여 1K01DK079019A2 (PT)에 의해 지원되었다.

1. 마우스 접종에 대한 박테리아 준비 다음은 무균 조건 하에서 수행되어야합니다. <ol><li> 모자와 하나 17 × 100 밀리미터 폴리 프로필렌 튜브를 타고 신선한 Luria 국물 (LB) 미디어 3 ML를 추가합니다. autoclaved 팁을 사용하면, 스트레인 CP1 2의 일부 냉동 박테리아 글리세롤 주식을 가지고 LB 미디어를 포함하는 관에 문화의 작은 수량을 전송할 수 있습니다. 산소는 여전히 튜브를 입력 할 수 있도록 튜브 캡을 교체 - 문화 에어로빅 조건 하에서 성장해야합니다. 37 번 장소 튜브 ° 하루 아침에 220 rpm으로 흔들어 보육의 C. </li><li> 그 다음날, 3 ML의 LB 미디어와 함께 새로운 튜브에 밤 문화의 5 μl를 전송하고 37 ° C.에서 보육 밤의 정적 조건 하에서 성장 </li><li> 날 AA 50 ML 튜브에 3 전송 문화의 40 μl는 40 ML의 LB 미디어 각을 포함. 37 ° C 정적 보육의 하룻밤의 장소. </li><li> 켜, 4 ° C.에 원심 분리기를 설정일단 원심 분리기 최종 온도에 도달, 40 ML Nalgene의 원심 분리기 튜브에 각각 40 ML 문화를 전송할 수 있습니다. </li><li> 필요에 따라 LB 미디어와 함께 볼륨을 조절 - 원심 분리기의 균형을 튜브를 무게. 20 분에 6,000 rpm으로 튜브를 봐. </li><li> 조심스럽게 표면에 뜨는 제거하고 부드럽게 40 ML 멸균 PBS에서 박테리아 펠렛을 일시 중지합니다. </li><li> PBS와 단계 1.5를 반복합니다. </li><li> 표면에 뜨는을 대기음, 500 μl 멸균 PBS에서 박테리아를 일시 중지합니다. 1.5 ML의 microfuge 관에 솔루션을 전송합니다. 정지 10 μl를 타고 얼음처럼 차가운 PBS의 990 μl에 희석. 접종에 필요한 볼륨을 확인하려면 OD 420의 NM을 읽어 분광 광도계를 사용합니다. 의 적절한 볼륨을 확인하려면 얼음처럼 차가운 PBS가 펠릿을 중지 할 수 : OD 420의 나노 번호 (ML)를 타고 박테리아 펠렛 [예의 예상 볼륨을 뺍니다. OD 420 nm 정도 = 0.545, 펠렛 약 0.075 ML. 0.545-0.075 = 0.470]. </li><li> 10 박테리아 정지 μl와 디를 제거990 μl 얼음처럼 차가운 PBS의 류트. OD 420의 나노를 참조하십시오. 목표는 1.000의 희석 정지 ± 0.010을위한 OD 420의 nm의 값을 달성하는 것입니다. 귀하의 희석의 수가이 목표를 초과하면, 정지에 더 많은 볼륨을 추가하고 또 다른 읽기를. 수 아래 인 경우 0.990는 원하는 반응을 얻을 때까지 중지 및 반복 OD 420의 나노 읽기 스핀 다운. </li></ol> 2. 동물 감염 마우스 (5-7주 세, 그룹 당 10)가 잭슨 연구소 (바 하버, ME)에서 구입했다. <ol><li> 마우스는 Plexiglas 챔버에서 isoflurane 1~4%의 흡입에 의한 anesthetized하고 휴식하고있는 때까지 모니터링한다. </li><li> 한 번에 하나의 마우스는 유리 해밀턴 주사기 (1.5-2.0 cm의 길이)의 수정 된 30 G 바늘에 부착 된 폴리에틸렌 (PE10) 카테터와 catheterization에 의해 박테리아의 기지에 이루어집니다. 카테터는 바늘의 허브에 삽입됩니다. 마우스 plac입니다에드는 마취상의 등쪽 표면에 코 콘을 사용하여 유지. </li><li> 마우스의 음경은 음경 요도의 입구의 윤활에 사용되는 부드러운 압력과 면봉에 윤활제의 자유 양에 의해 압출입니다. </li><li> 10 μl 인산염은 버퍼 1 개 10 8 박테리아를 포함하는 염분은 catheterization 다음 anesthetized 마우스의 요도에 소개되어 있습니다. </li><li> 세균 첨부 파일을 허용하고 즉각적인 배뇨을 방지하기 위해 세균 소개 한 후 마우스가 Plexiglas 챔버에서 15 분 동안 anesthetized 상태로 유지됩니다. </li><li> 마우스들은 케이지에 다시 배치하고 24 시간 모니터링하고 있습니다. </li></ol> 3. 동작 테스트 마우스는 사전 감염 (기준)과 일 3, 7, 14, 21, 감염 후 28 테스트되었습니다. 라고 hyperalgesia와 촉각 allodynia은 복부 11,12과 발바닥 R에 적용 폰 프라이의 필라멘트를 사용하여 테스트되었습니다뒷다리 앞발로 땅을 차다 13 egion. 테스트는 시간의 하루 고정에 perfomed 된 표준 방법론과 모든 동물의 한 실험 테스트는 고용했다. 그룹의 눈이 멀어 테스트는 동물 모델에서 동작 기반의 통증 테스트의 한계를 방지하기 위해 활용되었다. 0.04의 힘, 0.16, 0.4, 1.0 및 4.0 g (Stoelting, USA)와 다섯 개인 폰 프라이의 필라멘트가 탈퇴 응답의 복부 및 주파수에 적용되었다가 계산되었습니다. <ol><li> 새 환경 순응의 30 분 기간 후, 마우스는 스테인레스 스틸 와이어 그리드 바닥 - 집에서 만든 개인 Plexiglas 챔버 (6 × 10 x12 cm)에 배치됩니다. </li><li> 라고 hyperalgesia와 촉각 allodynia은 다섯 폰 프라이의 필라멘트를 사용하여 테스트되었습니다. 각 필라멘트는 총 10 회에 5 초의 상호 자극 간격으로 1-2 초에 대해 적용하고, 머리카락이 힘의 오름차순으로 테스트되었습니다. </li><li> 각 필라멘트는 전립선의 일반적인 주변에 낮은 복부 지역에 적용되고주의 desensitization을 피하​​거나 효과 &quot;를 맺어야&quot;이 지역 내의 다른 지역을 자극하는 촬영되었습니다. 필라멘트는 10 배의 총 stimulations 사이에 최소 5 초 간격으로 1-2 초에 대해 힘의 순서를 증가에 적용되었습니다. </li><li> 행동의 세 가지 유형은 필라멘트 자극에 긍정적 인 반응으로 간주됩니다 : 복부 1) 날카로운 철회, 2) 즉시 핥아 또는 필라멘트 자극의 지역을 긁적, 3) 점프. </li><li> 응답 주파수는 긍정적 인 반응의 비율 (10 중에 10 = 50 % 예를 들어 5 응답)로 계산 및 데이터가 응답 주파수의 평균 비율 ± SE로보고되었다 </li><li> 25 개 이상의 긍정적 총 기본 응답과 동물은 연구에서 제외됩니다. </li><li> 촉각 allodynia는 0.04의 힘, 0.16, 0.4, 1.0 및 4.0 g과 폰 프라이의 필라멘트를 사용하여 뒷다리 손 안에 발바닥 지역에서 테스트되었습니다. 중간 50 % 탈퇴 임계 값 (5)이 위, 아래를 사용하여 평가 한필라멘트가 약간 구부러 때까지 테스트가 0.04 g 필라멘트로 시작 방법은 뒷다리 손 안에 발바닥 표면에 수직 적용. 필라멘트는 긍정적 인 반응이 관찰 될 때까지 오름차순으로 테스트되었습니다. 필라멘트에 대한 긍정적 인 반응은 테스트 손 안에 앞발로 땅을 차다이나 핥아의 날카로운 철회 중 하나로 정의되었다. 긍정적 인 반응이 기록되었을 때 다음 약한 필라멘트가 적용되고, 부정적인 응답이 관찰 된 경우, 다음 강한 필라멘트가 적용되었습니다. </li><li> 설명 실험과 방법론은 검토 및 승인 노스 웨스턴 대학 동물 의료 의해위원회를 사용하고 있습니다. </li></ol>

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관심 없음 충돌이 선언 없습니다.

UPEC와 마우스 전립선의 감염은 세균 prostatitis, CP / CPPS의 pathogenesis 또는 만성 염증에 predisposing 이벤트로 참여 할 수 있습니다 이벤트의 생체 모델링에서 할 수 있습니다. 방법은 여성 비뇨기의 요로 감염이 7,14에 UPEC 모델에 문학의 큰 몸에 세균 준비 및 떨어 뜨림 조 추첨에 대해 설명했다. 모델은 잠재적 인 백신 후보와 면역 변조의 메커니즘을 테스트 pathogenesis을 공부에 대한 폭 넓...

<table border="1"><tbody><tr><td> 시약의 이름 </td><td> 회사 </td><td> 카탈로그 번호 </td></tr><tr><td> 문화 튜브 </td><td> BD 팔콘 </td><td> 352059 </td></tr><tr><td> LB 국물 밀러 </td><td> EMD </td><td> EM1.10285.0500 </td></tr><tr><td> Nalgene의 원심 분리기 튜브 </td><td> 열 </td><td> 3118-0050 </td></tr><tr><td> Isoflurane </td><td> 버틀러 Schein </td><td> NDC 11695-6776-1 </td></tr><tr><td> 카테 테르 바늘 30g의 </td><td> 해밀턴 </td><td> 91,030 </td></tr><tr><td> PE10 튜브 </td><td> BD intramedic </td><td> 427400 </td></tr><tr><td> 본 프라이의 필라멘트 </td><td> Stoelting </td><td> 58025-31 </td></tr></tbody></table>

measurement of tactile allodynia in a murine model of bacterial prostatitis

Uropathogenic 대장균 (UPEC)는 요로 감염 및 세균 prostatitis 1 중요한 역할을 병원균입니다. 우리는 최근에 UPEC는 만성 골반 통증 2, 만성 prostatitis / 만성 골반 통증 증후군 (CP / CPPS) 3,4의 기능의 개시에 중요한 역할을하는 것으로 나타났습니다. 임상 관련 UPEC하여 전립선의 감염은 조직 손상과 autoimmunity 5 메커니즘의 개시를 포함 할 수 있습니다 메커니즘을 통해 만성 통증을 시작하고 설정할 수 있습니다. 전립선에 UPEC의 pathogenesis를 이해하는 데 도전 조작에 전립선의 상대적 어려움입니다. 우리는이를 통해 전립선의 오름차순 감염을 수립 남성 생쥐에 UPEC의 임상 부담이 제공하는 이전에 설명 intraurethral 감염 방법 6을 활용. 여기, 우리는 학사 학위를 표준화에 대한 우리의 프로토콜을 설명뿐만 아니라 박테리아의 기지에 anesthetized 남성 쥐를 catheterizing 절차 등 cterial inoculum 7. CP / CPPS는 주로 촉각 allodynia (4)의 존재에 의해 특징입니다. 동작 테스트는 언급 내장 통증 8-10으로 인한 피부 hyperalgesia의 개념을 기반으로했다. 내장 조직에 짜증 초점은 일반적으로 비 고통스러운 자극 (allodynia)에 대한 과장된 반응을 허용하는 피부 통증 임계 값을 감소시킵니다. 기본 내장 고통의 강도를 증가하는 경향이 이상 반응의 피부 결과 정상적인 힘의 응용 프로그램입니다. 우리는 UPEC 박테리아와 하나의 감염에 대한 응답으로 시간이 지남에 촉각 allodynia를 정할 폰 프라이 섬유를 활용 NOD / ShiLtJ 마우스에 방법을 설명합니다.

<html> 우리는 세균 감염시 만성 통증의 존재에 대한 NOD / ShiLtj와 C57BL/6J 수컷 쥐를 조사했다. 남성 마우스 (5~7주 이전)은 요도 (그림 1)에 생리 나 세균과 배우는되었습니다. 생리 - 배우는 C57BL/6J 마우스와 UPEC에 감염된 C57BL/6J 마우스의 폰 프라이의 필라멘트와 골반 지역의 기계적 자극이 실험의 28 일 코스 (그림 1) 동안 변경되지 않은 응답 주파수하게되었습니다. 반면, UPEC에...

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

전립선의 감염은 만성 prostatitis의 골반 통증을 중재에 기여 요소가 될 수 있습니다. 우리는 표준화 된 박테리아 inoculum, 시간이 지남에 따라 마우스에 촉각 allodynia을 측정하는 남성 마우스 및 방법론의 요도에 박테리아의 기지의 준비에 대한 절차를 설명합니다.

세균 Prostatitis의 손쥐 모델에 촉각 Allodynia 측정

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

Research

JoVE Journal

Immunology and Infection

15.8K 조회수.  Northwestern University Feinberg School of Medicine. 전립선의 감염은 만성 prostatitis의 골반 통증을 중재에 기여 요소가 될 수 있습니다. 우리는 표준화 된 박테리아 inoculum, 시간이 지남에 따라 마우스에 촉각 allodynia을 측정하는 남성 마우스 및 방법론의 요도에 박테리아의 기지의 준비에 대한 절차를 설명합니다.

비디오: 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.

쥐에 있는 복잡한 지역 고통 증후군 타입-I를 위한 만성 포스트 허혈 고통 모형

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

연구

행동분석학

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.

마우스의 신경 병증 성 통증의 좌골 신경의 바지를 접 모델

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

의학

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 ...

신경과학

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.

드로소필라 애벌레의 기계적 고치절 측정을 위한 향상된 분석 및 도구

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.

열 및 기계를 침해 수용성 모달리티를 사용하여 마우스의 모르핀에 의한 통각 과민과 진통 공차의 평가

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.

만성 요통의 기계론적 조사를 위한 쥐의 등 기계적 민감도 평가

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.

우로키나아제(Urokinase)형 플라스미노겐 활성제(Plasminogen Activator)에 의한 생쥐 요통 모델

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.

신생아 모자 분리의 마우스 모델에서 Perigenital 감도 및 전립선 비만 세포 활성화의 평가

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.

성인 남성 마우스 transurethral 주입 절차

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

면역학 및 감염병학

T. evansi에 감염된 동물의 통증 및 이질증 평가

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.

저자 스포트라이트: 트리파노소마 에반시(Trypanosoma evansi)에 감염된 실험실 동물의 통증 평가 및 동물 복지 증진

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.

Trypanosoma evansi-Infected Mice의 Allodynia 및 통증을 평가하기 위해 Mouse Grimace Scale과 관련된 Electronic von Frey

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 ...