Name: chronic constriction of the sciatic nerve and pain hypersensitivity testing in rats
Uploaded: 2012-03-13T12:00:00.000+00:00
Duration: 8 min 23 s
Description: Watch this Scientific Journal Video about Chronic Constriction of the Sciatic Nerve and Pain Hypersensitivity Testing in Rats at JoVE.com

Os autores gostariam de agradecer a descrição original do modelo de CCI por Bennett e Xie (1988). Este trabalho foi financiado parcialmente por concessões do National Health and Medical Research Council da Austrália (ID # 568637) e do Instituto NSW para a Ciência e Investigação Médica ao GMT.

1. Constrição do nervo ciático crônica Técnicas assépticas deve ser utilizado para o procedimento cirúrgico. Desinfectar a superfície de trabalho cirúrgico com etanol a 70% e em preparar Advance Instruments estéril (por exemplo, tesouras finas, fórceps e afastador), gazes, grampos e esfregaços por autoclavagem. Para cirurgias múltiplas, limpar e instrumentos reesterilizar com etanol a 70% ou um esterilizador seco grânulo entre ratos. Uma máscara cirúrgica, gorro cabelo e luvas estéreis devem ser usadas. Para constrição do nervo, preparar a sutura intestino crómico cortando-o em pequenos pedaços de cerca de 3 cm de comprimento imersos em solução salina estéril, para evitar a secagem. Para fechar a ferida, usar comercialmente disponíveis suturas de seda estéreis. <ol><li> Anestesiar o rato em uma câmara de indução usando 5% de isoflurano em O 2, em seguida, entregar 2% de isoflurano em O 2 através de uma máscara facial feitos, e monitorar o animal anestesiado durante o procedimento. Anestésicos comuns alternativos são inalados halotano ou pentobarbital sódico intraperitoneal. A cetamina, como um antagonista do receptor N-metil-D-aspartato (NMDA), deve ser evitado uma vez que estes receptores são importantes no desenvolvimento de dor neuropática. </li><li> Raspar a pata traseira esquerda do rato. Em seguida, coloca o animal sobre uma esteira de aquecimento termo-regulada a 37 ° C e aplicar pomada oftálmica lubrificante para os olhos. </li><li> Esterilizar a área rapada com três aplicações alternadas de 70% de álcool isopropílico e uma solução de iodo. </li><li> Com o rato deitado no seu peito / tórax, elevar a perna esquerda e segure na posição com o Femor a 90 ° para a coluna usando fita adesiva no pé. </li><li> Fazer uma incisão na pele paralelamente, mas 3-4 mm abaixo da Femor, ea pele do livre a partir do músculo circundante da incisão através do corte através do tecido conjuntivo. </li><li> Com uma tesoura sem corte, cortam o tecido conjuntivo entre os glúteos superficial e os bícepsfemoris. </li><li> Utilize um afastador para alargar o fosso entre os dois músculos, permitindo a visualização clara do nervo ciático. </li><li> Usando curvas atraumáticas, fórceps e micro-tesouras, suavemente cerca de 10 mm livre do nervo ciático (proximal à trifurcação ciática) a partir do tecido conjuntivo circundante. </li><li> Sob o microscópio e uma boa fonte de luz, amarrá quatro ligaduras (crómico intestino 4,0, Ethicon, EUA) com um nó duplo, 1 mm entre si, proximal à trifurcação do nervo ciático. Para cada ligadura, comece com um único laço solto, segure as duas extremidades perto do loop e apertar até que o loop é apenas um pouco confortável ea ligadura não deslizar ao longo do nervo. Para segurar o lacete na sua posição correcta, colocar uma segunda alça no topo da primeira para completar o nó. Por fim, corte as pontas soltas da ligadura para cerca de 1mm. Constrição do nervo deve ser mínima, e imediatamente interrompido se uma contração breve é ​​observado, para evitar a prisão de the fluxo de sangue epineural. O aperto excessivo das ligaduras leva a axotomia e autotomia (auto-mutilação), ambos os indesejados efeitos colaterais que impedem teste de hipersensibilidade sucesso dor. </li><li> Use suturas (Mersilk 5,0, Ethicon) para fechar a camada muscular e grampos (Autoclip, 9 mm) para prender a pele. Em seguida, utilizar solução de iodo (Riodine) para esterilizar a ferida. </li><li> Observar o rato intimamente durante o período de recuperação da anestesia, e permitir que ele se recuperar numa gaiola separada com cama de papel plano (não o padrão da cama criação animal), a fim de impedir que o animal inconsciente asfixia. Antes de dar qualquer investigadores analgesia pós-operatória deve considerar o fato que pode interferir com o teste de hipersensibilidade à dor. De facto, há evidências de que a aplicação local de anestésico, lidocaína, no momento da lesão pode alterar o desenvolvimento de dor neuropática, e diminuir o grau de hiperalgesia nas semanas anteriores 7. Para mais discussão sobre esse mAtter ver revisão por Bennett et al 8. </li></ol> 2. Medidas de Comportamentos Dor <ol><li> Manipulação diária de ratos durante vários dias antes do início do experimento é recomendado para acostumar os animais ao seu manipulador, reduzir o estresse eo risco de morder. </li><li> Antes dos testes comportamentais, os ratos têm de ser habituados ao procedimento de teste e do invólucro animal. O ambiente de teste deve ser mantido calmo e bem controlado, com temperatura constante e os níveis de umidade e realizado em um tempo similar de dias de cada sessão de teste. </li><li> Teste de hipersensibilidade a dor deve ser realizada várias vezes antes de ICC (linha de base) e em pontos de tempo diferentes após ICC, como desejado. </li></ol> 3. Limiar de retirada mecânica <ol><li> No dia do teste, os ratos colocá-las no elevada gaiola de testes de malha floored 15-30 minutos antes de medir limiares de retirada, permitindo gaiola de exploração e Grooatividades ming para cessar e ratos para tornar-se relaxado e parado. </li><li> Usando uma dinâmica plantar von Frey aesthesiometer (Ugo Basile, Itália), a posição da unidade de contato estimulador com o filamento diretamente abaixo da superfície meados de plantar da pata traseira, evitando a malha de metal, e pressione a tecla start. Este levanta o filamento para mecanicamente estimular a superfície plantar da pata posterior do com uma força crescente. </li></ol> Cabelos de von Frey manuais são uma série de filamentos aumentando de diâmetro que são calibrados para induzir uma força específica quando aplicado à pele. Eles podem ser utilizados como uma alternativa para uma electrónica plantar von Frey dinâmica aesthesiometer para a medição de limiar de retirada da pata, como descrito anteriormente 4. Existem também vários fabricantes alternativos de von Frey eletrônico aesthesiometers (por exemplo IITC, EUA e Somedic, Suécia). <ol start="3"><li> Registre o limiar de retirada mecânica de hindpa tanto lesadas ews, em ordem contrabalanceada. Este dispositivo aplica-se pressão para a pata e após reacção de retirada o sistema automaticamente grava e exibe a força máxima aplicada (em gramas). Se uma resposta ambígua ocorre (isto é, no caso de deambulação ou preparação), repetir o estímulo e gravar o limiar de novo. Lamber ou agitação da estimulação seguinte pata é um sinal de comportamento associadas à dor. </li><li> Repita a estimulação mecânica 3 vezes com um intervalo de cerca de 5 min entre os estímulos, registrar os limiares, e depois calcular a média dos limiares de retirada da pata. </li></ol> 4. Latência Retirada térmica A intensidade de calor de infravermelhos (IR) do instrumento de teste plantar deve ser ajustado antes de iniciar a experiência. Em nosso laboratório, nós ajustar a intensidade IR para dar uma latência média de abandono pata de cerca de 10 segundos em ratos ingênuos, e vamos definir a latência de corte, 20 segundos para evitar danos tecidoidade em caso de falha para remover uma pata. <ol><li> No dia do teste, os ratos lugar no interior da gaiola de testes de vidro com piso de 15-30 minutos antes de medir a latência, permitindo a exploração gaiola e tosa actividades de cessar e ratos para se tornar relaxado e estacionários. </li><li> Usando um instrumento de analgesia térmica plantar (Ugo Basile, Itália), a posição da fonte de infravermelho diretamente abaixo da superfície meados de plantar da pata traseira, e pressione a tecla start. Isto expõe a superfície plantar da pata posterior do a um feixe de calor radiante através de uma superfície de vidro transparente. Há também fabricantes alternativos de térmicas instrumentos de analgesia plantar (por exemplo, IITC, EUA e Stoelting, EUA). </li><li> Registre a latência de retirada de patas traseiras tanto lesadas e não lesadas, em ordem contrabalanceada. Este dispositivo regista automaticamente o tempo decorrido (em segundos) a partir do início do estímulo térmico para a retirada da pata da fonte de calor. Se uma resposta ambígua ocorre (isto é, no caso de ambulação ou grooming), repita o estímulo e gravar a latência novamente. Lamber ou agitação da estimulação seguinte pata é um sinal de comportamento associadas à dor. </li><li> Repita a estimulação de calor, pelo menos, 3 vezes com um intervalo de cerca de 5 min entre estímulos, gravar as latências e, em seguida o cálculo da média das latências de retirada da pata. </li></ol> 4. Os resultados representativos Um exemplo de limiar de retirada mecânica e latência de retirada térmica nas patas ipsilateral (lesionado) e contralateral (não lesionado) de ratos Wistar é mostrado antes (linha de base) e 2-12 dias após ICC na Figura 3A, e Figura 3B. A partir destes dados pode ser visto que a CCI, provoca uma redução significativa em ambos os limiares de retirada mecânicas e as latências de retirada térmicas da pata posterior ipsilateral, indicando a presença de hipersensibilidade dor mecânica e térmica, ambas as características de dor neuropática crónica<html> . <img alt="A Figura 1" src="/files/ftp_upload/3393/3393fig1.jpg" /> Figura 1. Um esquemático que ilustra o posicionamento das ligaduras no modelo de ICC. Este modelo animal de dor neuropática envolve a colocação de quatro soltas crômico intestino-ligaduras sobre o nervo ciático, no meio da coxa nível. Isso resulta em ferindo apenas uma porção das fibras aferentes que vão para o hipersensibilidade dor no pé e subsequente da pata posterior. <img alt="A Figura 2" src="/files/ftp_upload/3393/3393fig2.jpg" /> Figura 2. Diagrama da área pata posterior utilizado para testes de dor. A superfície meados de plantar apenas posterior para as patas, como indicado pelo círculo vermelho pontilhado, é utilizado para testes de dor. Esta área é inervada principalmente pelos ramos terminais do nervo ciático. O calcanhar e patas devem ser evitados devido à sensibilidade da pele diferencial. 3.jpg &quot;/&gt; Figura hipersensibilidade Dor 3. Após constrição crônica do nervo ciático em ratos. Depois de ICC, há uma redução significativa no limiar de retirada da pata a estímulos mecânicos (A) e latência de retirada da pata a estímulos térmicos (B) na ferida (ipsilateral) pata posterior comparação com o não lesionado (contralateral) pata posterior (P &lt;0,01 medidas repetidas 2 -way ANOVA). Os dados mostram média ± EPM, n = 7 ratos Wistar.

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Neurosci. 10, 283-294 (2009).</li><li>Langford, D. J. <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> Nat. Methods. 7, 447-449 (2010).</li><li>Austin, P. J., Moalem-Taylor, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+neuro-immune+balance+in+neuropathic+pain:+involvement+of+inflammatory+immune+cells,+immune-like+glial+cells+and+cytokines.">The neuro-immune balance in neuropathic pain: involvement of inflammatory immune cells, immune-like glial cells and cytokines.</a> J. Neuroimmunol. 229, 26-50 (2010).</li></ol>

Experimentos com animais foram aprovados pelo Animal Care e Comitê de Ética da Universidade de New South Wales, Austrália, e seguiu as orientações formuladas pela Associação Internacional para o Estudo da Dor.

CCI é amplamente utilizado modelo de lesão de nervo periférico da dor neuropática crônica. É relativamente simples de executar, e produz hipersensibilidade dor robusta e estável durante pelo menos um mês após a lesão. Após CCI, os ratos apresentam postura anormal da pata ferida (dedos unidos e plantar-flexionados e pata evertido), bem como repetidos tremores, guarda e lambendo da pata ferida sugerindo a presença de dor espontânea 9. Além disso a disfunção sensorial, vários investigadores demo...

<table border="1"><tbody><tr><td> Nome do reagente / equipamentos </td><td> Companhia </td><td> Número de catálogo </td><td> Comentários </td></tr><tr><td> Gut crômico </td><td> Ethicon </td><td> G-211 </td><td> Espessura 4-0 </td></tr><tr><td> Isoflurano </td><td> Delvet Pty. Ltd., Sete Colinas, NSW </td><td> n / a </td><td></td></tr><tr><td> Mersilk </td><td> Ethicon </td><td> W-580 </td><td> 5-0 espessura </td></tr><tr><td> Autoclip </td><td> Becton Dickinson </td><td> 427631 </td><td> 9 milímetros de aço inoxidável </td></tr><tr><td> Riodine </td><td> Orion </td><td> R1000802F </td><td> 1% de iodo w / v </td></tr><tr><td> Instrumento analgesia térmica plantar </td><td> Ugo Basile </td><td> 37370 </td><td></td></tr><tr><td> Aesthesiometer plantar dinâmica </td><td> Ugo Basile </td><td> 37400 </td><td></td></tr></tbody></table>

chronic constriction of the sciatic nerve and pain hypersensitivity testing in rats

Dor neuropática crônica, resultante de danos ao sistema nervoso central ou periférico, é uma condição prevalente e incapacitante, afetando 7-18% da população 1,2. Os sintomas incluem dor (queimação, formigamento, choque elétrico como) espontânea, disestesia, parestesia, alodinia (dor resultante de normalmente não dolorosos estímulos) e hiperalgesia (um aumento de resposta a estímulos dolorosos). Os sintomas sensoriais são co-mórbidas com deficiências comportamentais, tais como insônia e depressão. Para estudar a dor neuropática crónica vários modelos animais que imitam a lesão do nervo periférico têm sido desenvolvidos, um dos mais amplamente utilizado é Bennett e (1988) unilateral de Xie lesão do nervo ciático constrição crónica (ICC) 3 (Figura 1). Aqui apresentamos um método para executar ICC e teste de hipersensibilidade a dor. ICC é realizada sob anestesia, com o nervo ciático de um lado exposto fazendo comoincisão parentes, e cortando o tecido conjuntivo entre as superficial glúteos e bíceps femoral. Quatro ligaduras intestino crômico são amarrados frouxamente ao redor do nervo ciático em intervalos de 1 mm, apenas para ocluir, mas não deter o fluxo de sangue epineural. A ferida é fechada com suturas no músculo e grampos na pele. O animal é então permitido recuperar da cirurgia durante 24 horas antes do teste de hipersensibilidade dor começa. Para os testes comportamentais, os ratos são colocados no dispositivo de ensaio e são permitidos para habituar o procedimento de teste. A área de teste é a superfície mid-plantar da pata posterior (Figura 2), que cai dentro da distribuição do nervo ciático. Limiar de retirada mecânica é avaliado por ambas as patas traseiras mecanicamente estimulando lesadas e usando um eletrônico dinâmico plantar von Frey aesthesiometer ou manual von Frey cabelos 4. O limiar de retirada mecânica é a pressão máxima exercida(Em gramas) que desencadeia retirada da pata. Para a medição da latência de retirada térmica, primeiro descrito por Hargreaves et ai (1988), a pata posterior é exposta a um feixe de calor radiante através de uma superfície de vidro transparente com um medidor de analgesia plantar 5,6. A latência de retirada ao estímulo de calor é registrado como o tempo para retirada da pata nas duas patas traseiras lesadas e não lesadas. ICC sequência, limiar de retirada mecânica, bem como a latência de retirada térmica na pata lesada são ambos significativamente reduzido, em comparação com as medições de linha de base e da pata não lesionado (Figura 3). O modelo ICC de lesão do nervo periférico combinados com testes de hipersensibilidade à dor fornece um sistema modelo para investigar a eficácia de agentes terapêuticos potenciais para modificar a dor neuropática crônica. No nosso laboratório, nós utilizamos ICC ao lado de sensibilidade térmica e mecânica das patas traseiras para investigar o papel das interacções neuro-imunes na patogénesee tratamento da dor neuropática.

Watch this Scientific Journal Video about Chronic Constriction of the Sciatic Nerve and Pain Hypersensitivity Testing in Rats at JoVE.com

Chronic Constriction of the Sciatic Nerve and Pain Hypersensitivity Testing in Rats

Devido à simplicidade da cirurgia e os resultados robusto comportamental, constrição crónica do nervo ciático é um dos modelos pré-eminentes animais de dor neuropática. Dentro de 24 horas após a cirurgia, hipersensibilidade à dor é estabelecido e pode ser medida quantitativamente utilizando um von Frey aesthesiometer (teste mecânico) e medidor de analgesia plantar (teste térmico).

Constrição crônica do nervo ciático e Teste de Hipersensibilidade dor em ratos

Chronic neuropathic pain, resulting from damage to the central or peripheral nervous system, is a prevalent and debilitating condition, affecting 7-18% of ...

chronic-constriction-sciatic-nerve-pain-hypersensitivity-testing

Research

JoVE Journal

Medicine

58.7K visualizações.  University of New South Wales. Devido à simplicidade da cirurgia e os resultados robusto comportamental, constrição crónica do nervo ciático é um dos modelos pré-eminentes animais de dor neuropática. Dentro de 24 horas após a cirurgia, hipersensibilidade à dor é estabelecido e pode ser medida quantitativamente utilizando um von Frey aesthesiometer (teste mecânico) e medidor de analgesia plantar (teste térmico).

Vídeo: Chronic Constriction of the Sciatic Nerve and Pain Hypersensitivity Testing in Rats

The Spared Nerve Injury (SNI) Model of Induced Mechanical Allodynia in Mice

Peripheral neuropathic pain is a severe chronic pain condition which may result from trauma to sensory nerves in the peripheral nervous system. The spared nerve injury (SNI) model induces symptoms of neuropathic pain such as mechanical allodynia i.e. pain due to tactile stimuli that do not normally provoke a painful response [1]. 
The SNI mouse model involves ligation of two of the three branches of the sciatic nerve (the tibial nerve and the common peroneal nerve), while the sural nerve is left intact [2]. The lesion results in marked hypersensitivity in the lateral area of the paw, which is innervated by the spared sural nerve. The non-operated side of the mouse can be used as a control. The advantages of the SNI model are the robustness of the response and that it doesn&#x2019;t require expert microsurgical skills.
The threshold for mechanical pain response is determined by testing with von Frey filaments of increasing bending force, which are repetitively pressed against the lateral area of the paw [3], [4]. A positive pain reaction is defined as sudden paw withdrawal, flinching and/or paw licking induced by the filament. A positive response in three out of five repetitive stimuli is defined as the pain threshold. 
As demonstrated in the video protocol, C57BL/6 mice experience profound allodynia as early as the day following surgery and maintain this for several weeks.

The Spared Nerve Injury SNI Model of Induced Mechanical Allodynia in Mice

The Spared Nerve Injury animal model is described here as a mouse model of peripheral neuropathic pain following partial denervation of the sciatic nerve by lesioning the tibial and common peroneal nerve branches, leaving the remaining sural nerve intact. Behavioral modification resulting from mechanical allodynia is quantified by von Frey filaments.

Poupado da lesão do nervo Model (SNI) de alodinia mecânica induzida em camundongos

Peripheral neuropathic pain is a severe chronic pain condition which may result from trauma to sensory nerves in the peripheral nervous system. The spared ...

Medicina

Acute and Chronic Tactile Sensory Testing after Spinal Cord Injury in Rats

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory testing in rat SCI models is needed. However, until recently, no single testing approach had been validated for SCI so that standardized methods have not been implemented across labs. Additionally, available testing methods could not be implemented acutely or when severe motor impairments existed, preventing studies of the development of SCI-induced allodynia3. Here we present two validated sensory testing methods using von Frey Hair (VFH) monofilaments which quantify changes in tactile sensory thresholds after SCI4-5. One test is the well-established Up-Down test which demonstrates high sensitivity and specificity across different SCI severities when tested chronically5. The other test is a newly-developed dorsal VFH test that can be applied acutely after SCI when allodynia develops, prior to motor recovery4-5. Each VFH monofilament applies a calibrated force when touched to the skin of the hind paw until it bends. In the up-down method, alternating VFHs of higher or lower forces are used on the plantar L5 dermatome to delineate flexor withdrawal thresholds. Successively higher forces are applied until withdrawal occurs then lower force VFHs are used until withdrawal ceases. The tactile threshold reflects the force required to elicit withdrawal in 50% of the stimuli. For the new test, each VFH is applied to the dorsal L5 dermatome of the paw while the rat is supported by the examiner. The VFH stimulation occurs in ascending order of force until at least 2 of 3 applications at a given force produces paw withdrawal. Tactile sensory threshold is the lowest force to elicit withdrawal 66% of the time. Acclimation, testing and scoring procedures are described. Aberrant trials that require a retest and typical trials are defined. Animal use was approved by Ohio State University Animal Care and Use Committee.

We describe two tactile sensory testing methods for acute or chronic periods of spinal cord injury in rats. These validated procedures can detect the development and maintenance of allodynia-like sensations.

Aguda e crônica teste sensorial tátil após a lesão medular em ratos

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory ...

Ascending Aortic Constriction in Rats for Creation of Pressure Overload Cardiac Hypertrophy Model

Ascending aortic constriction is the most common and successful surgical model for creating pressure overload induced cardiac hypertrophy and heart failure. Here, we describe a detailed surgical procedure for creating pressure overload and cardiac hypertrophy in rats by constriction of the ascending aorta using a small metallic clip. After anesthesia, the trachea is intubated by inserting a cannula through a half way incision made between two cartilage rings of trachea. Then a skin incision is made at the level of the second intercostal space on the left chest wall and muscle layers are cleared to locate the ascending portion of aorta. The ascending aorta is constricted to 50&#8211;60% of its original diameter by application of a small sized titanium clip. Following aortic constriction, the second and third ribs are approximated with prolene sutures. The tracheal cannula is removed once spontaneous breathing was re-established. The animal is allowed to recover on the heating pad by gradually lowering anesthesia. The intensity of pressure overload created by constriction of the ascending aorta is determined by recording the pressure gradient using trans-thoracic two dimensional Doppler-echocardiography. Overall this protocol is useful to study the remodeling events and contractile properties of the heart during the gradual onset and progression from compensated cardiac hypertrophy to heart failure stage.

We describe a stepwise procedure for creating pressure overload and left ventricular hypertrophy in Wistar rats by constriction of the ascending aorta using a small metallic clip. This model is extensively used for studying remodeling changes during cardiac hypertrophy and for identifying and evaluating strategies for regression of such changes.

Aorta ascendente constrição em ratos para a criação de sobrecarga de pressão Cardíaca Hipertrofia Modelo

Ascending aortic constriction is the most common and successful surgical model for creating pressure overload induced cardiac hypertrophy and heart ...

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.

Modelo crônico da dor da Borne-Isquemia para a síndrome regional complexa da dor tipo-I nos ratos

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

Comportamento

Urinary Bladder Distention Evoked Visceromotor Responses as a Model for Bladder Pain in Mice

Approximately 3-8 million people in the United States suffer from interstitial cystitis/bladder pain syndrome (IC/BPS), a debilitating condition characterized by increased urgency and frequency of urination, as well as nocturia and general pelvic pain, especially upon bladder filling or voiding. Despite years of research, the cause of IC/BPS remains elusive and treatment strategies are unable to provide complete relief to patients. In order to study nervous system contributions to the condition, many animal models have been developed to mimic the pain and symptoms associated with IC/BPS. One such murine model is urinary bladder distension (UBD). In this model, compressed air of a specific pressure is delivered to the bladder of a lightly anesthetized animal over a set period of time. Throughout the procedure, wires in the superior oblique abdominal muscles record electrical activity from the muscle. This activity is known as the visceromotor response (VMR) and is a reliable and reproducible measure of nociception. Here, we describe the steps necessary to perform this technique in mice including surgical manipulations, physiological recording, and data analysis. With the use of this model, the coordination between primary sensory neurons, spinal cord secondary afferents, and higher central nervous system areas involved in bladder pain can be unraveled. This basic science knowledge can then be clinically translated to treat patients suffering from IC/BPS.

Approximately 3-8 million people in the United States suffer from interstitial cystitis/bladder pain syndrome (IC/BPS), a debilitating condition characterized in part by pelvic pain. In order to study nervous system contributions to the condition, a physiological model of urinary bladder pain is used in mice and rats.

Respostas Bexiga Distensão Evocado Visceromotor como um modelo para Bexiga dor em camundongos

Approximately 3-8 million people in the United States suffer from interstitial cystitis/bladder pain syndrome (IC/BPS), a debilitating condition ...

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.

O ciático Nervo cuffing Modelo de Dor Neuropática em Ratos

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

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.

Avaliação da Sensibilidade Mecânica das Costas no Rato para Investigação Mecanicista da Dor Lombar Crônica

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

Partial Sciatic Nerve Ligation: A Mouse Model of Chronic Neuropathic Pain to Study the Antinociceptive Effect of Novel Therapies

Management of chronic pain remains challenging to this day, and current treatments are associated with adverse effects, including tolerance and addiction. Chronic neuropathic pain results from lesions or diseases in the somatosensory system. To investigate potential therapies with reduced side effects, animal pain models are the gold standard in preclinical studies. Therefore, well-characterized and well-described models are crucial for the development and validation of innovative therapies. 
Partial ligation of the sciatic nerve (pSNL) is a procedure that induces chronic neuropathic pain in mice, characterized by mechanical and thermal hypersensitivity, ongoing pain, and changes in limb temperature, making this model a great fit to study neuropathic pain preclinically. pSNL is an advantageous model to study neuropathic pain as it reproduces many symptoms observed in humans with neuropathic pain. Furthermore, the surgical procedure is relatively fast and straightforward to perform. Unilateral pSNL of one limb allows for comparison between the ipsilateral and contralateral paws, as well as evaluation of central sensitization. 
To induce chronic neuropathic hypersensitivity, a 9-0 non-absorbable nylon thread is used to ligate the dorsal third of the sciatic nerve. This article describes the surgical procedure and characterizes the development of chronic neuropathic pain through multiple commonly used behavioral tests. As a plethora of innovative therapies are now being investigated to treat chronic pain, this article provides crucial concepts for standardization and an accurate description of surgeries required to induce neuropathic pain.

Partial sciatic nerve ligation induces long-lasting chronic neuropathic pain, characterized by exaggerated responses to thermal and mechanical stimuli. This mouse model of neuropathic pain is commonly used to study innovative therapies for pain management. This article describes in detail the surgical procedure to improve standardization and reproducibility.

Ligadura parcial do nervo ciático: um modelo murino de dor neuropática crônica para estudar o efeito antinociceptivo de novas terapias

Management of chronic pain remains challenging to this day, and current treatments are associated with adverse effects, including tolerance and addiction. ...

<meta charset="utf8"></head><body>Estabelecendo o modelo de camundongo para dor neuropática do trigêmeo

Chronic Constriction Injury of the Distal Infraorbital Nerve (DIoN-CCI) in Mice to Study Trigeminal Neuropathic Pain

Animal models remain necessary tools to study neuropathic pain. This manuscript describes the distal infraorbital nerve chronic constriction injury (DIoN-CCI) model to study trigeminal neuropathic pain in mice. This includes the surgical procedures to perform the chronic constriction injury and the postoperative behavioral tests to evaluate the changes in spontaneous and evoked behavior that are signs of ongoing pain and mechanical allodynia. The methods and behavioral readouts are similar to the infraorbital nerve chronic constriction injury (IoN-CCI) model in rats. However, important changes are necessary for the adaptation of the IoN-CCI model to mice. First, the intra-orbital approach is replaced by a more rostral approach with an incision between the eye and the whisker pad. The IoN is thus ligated distally outside the orbital cavity. Secondly, due to the higher locomotor activity in mice, allowing rats to move freely in small cages is replaced by placing mice in custom-designed and constructed restraining devices. After DIoN ligation, mice exhibit changes in spontaneous behavior and in response to von Frey hair stimulation that are similar to those in IoN-CCI rats, i.e., increased directed face grooming and hyperresponsiveness to von Frey hair stimulation of the IoN territory.

<meta charset="utf8"></head><body>Autor em destaque: Utilizando a ligadura do nervo infraorbital em camundongos para investigar a dor neuropática do trigêmeo e estratégias de tratamento

Chronic constriction injury of the distal infraorbital nerve in mice induces changes in spontaneous behavior (increased face grooming activity) and nocifensive behavior in response to tactile stimulation (hyperresponsiveness to von Frey hair stimulation) that are signs of ongoing pain and allodynia and serves as a model for trigeminal neuropathic pain.

Lesão por Constrição Crônica do Nervo Infraorbital Distal (DIoN-CCI) em Camundongos para Estudo da Dor Neuropática do Trigêmeo

Animal models remain necessary tools to study neuropathic pain. This manuscript describes the distal infraorbital nerve chronic constriction injury ...

Constrição crônica do nervo ciático e Teste de Hipersensibilidade dor em ratos

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