Los autores desean agradecer a la descripción original de la ICC modelo de Bennett y Xie (1988). Este trabajo fue parcialmente financiado por subvenciones de la Nacional de Salud y Consejo de Investigación Médica de Australia (ID # 568637) y la Oficina de NSW para la Ciencia y la Investigación Médica a la hora GMT.

1. La constricción del nervio ciático crónica Técnicas asépticas debe ser utilizado para el procedimiento quirúrgico. Desinfectar la superficie de trabajo quirúrgico con 70% de etanol y preparar los instrumentos estériles por adelantado (por ejemplo, tijeras, pinzas finas y retractores), gasas, grapas y los hisopos en autoclave. Para múltiples cirugías, limpieza y re-esterilizar instrumentos con etanol al 70% o un esterilizador seco cordón entre las ratas. Una máscara quirúrgica, sombrero de pelo y guantes estériles deben ser usados. Para la constricción del nervio, preparar la sutura intestino crómico cortándolo en trozos pequeños de aproximadamente 3 cm de longitud sumergido en solución salina estéril, para evitar el secado. Para cerrar la herida, uso de suturas estériles disponibles en el mercado de la seda. <ol><li> Anestesiar la rata en una cámara de inducción con isoflurano en un 5% de O 2, y luego entregar isoflurano 2% de O 2 a través de un encargo a la mascarilla, y vigilar el animal anestesiado durante el procedimiento. Comunes anestésicos alternativos son inhalados halotano o sodio pentobarbital intraperitoneal. La ketamina, como un antagonista del receptor N-metil-D-aspartato (NMDA), debe evitarse ya que estos receptores son importantes en el desarrollo del dolor neuropático. </li><li> Afeitado de la pierna izquierda trasera de la rata. Luego, coloque el animal en una rejilla de calefacción termo-regulado a 37 ° C y aplique una pomada oftálmica lubricante para los ojos. </li><li> Esterilizar la zona afeitada con tres aplicaciones alternas de 70% de alcohol isopropílico y solución de yodo. </li><li> Con la rata acostada en su pecho / tórax, eleve la pierna trasera izquierda y mantener en posición con el Femor a 90 ° de la columna vertebral utilizando cinta adhesiva en el pie. </li><li> Hacer una incisión en la piel paralelo, pero 3-4 mm por debajo de la Femor, y libre de la piel del músculo que rodea la incisión cortando a través del tejido conectivo. </li><li> Usando tijeras de punta roma, cortados a través del tejido conectivo entre los glúteos y bíceps superficial de losfemoris músculos. </li><li> Utilice un retractor para ampliar la brecha entre los dos músculos, lo que permite una clara visualización del nervio ciático. </li><li> Utilizando curvas de punta roma pinzas y tijeras microempresas, con suavidad libre de aproximadamente 10 mm del nervio ciático (proximal a la trifurcación ciática) del tejido conectivo circundante. </li><li> Bajo el microscopio y una buena fuente de luz, atar cuatro ligamentos (crómico intestino 4.0, Ethicon, EE.UU.) con un nudo doble, 1 mm de distancia, próximo a la trifurcación del nervio ciático. Para cada ligadura, comenzar con una sola vuelta suelta, y sujete los dos extremos cercanos al bucle y apretar hasta que el bucle está apenas ajustado y la ligadura no se desliza a lo largo del nervio. Para mantener el bucle en su posición correcta, colocar un segundo bucle en la parte superior de la primera para completar el nudo. Por último, cortar los cabos sueltos de la ligadura a alrededor de 1 mm. La constricción del nervio debe ser mínima, y ​​se detuvo de inmediato si una contracción breve se observa, para evitar la detención de THe flujo sanguíneo epineural. Si se aprieta demasiado las ligaduras conduce a axotomía y autotomía (automutilación), ambos efectos secundarios no deseados que se opongan a las pruebas de hipersensibilidad con éxito el dolor. </li><li> Use suturas (5,0 Mersilk, Ethicon) para cerrar la capa muscular y grapas (autoclip, 9 mm) para sujetar la piel. A continuación, utilice una solución de yodo (Riodine) para esterilizar la herida. </li><li> Tenga en cuenta que la rata de cerca durante el período de recuperación de la anestesia, y permite que se recupere en una jaula por separado con ropa de cama plana de papel (no en la ropa de cama de la cría de animales de serie), con el fin de evitar que el animal está inconsciente asfixia. Antes de dar cualquier investigación de la analgesia postoperatoria debe considerar el hecho de que esto puede interferir con las pruebas de dolor de hipersensibilidad. De hecho, hay evidencia de que la aplicación local de anestésico, lidocaína, en el momento de la lesión puede alterar el desarrollo del dolor neuropático, y disminuir el grado de hiperalgesia en las semanas anteriores 7. Para una mayor discusión sobre este mAtter ver la revisión de Bennett et al 8. </li></ol> 2. Las mediciones de conductas de dolor <ol><li> La manipulación diaria de las ratas durante varios días antes de iniciar el experimento, se recomienda con el fin de acostumbrar a las ratas a su manejador, reducir el estrés y el riesgo de morder. </li><li> Antes de las pruebas de comportamiento, las ratas que se habituaron al procedimiento de prueba y el recinto para animales. El entorno de prueba debe mantenerse tranquila y bien controlada, con una temperatura constante y los niveles de humedad, y lleva a cabo a la misma hora del día de cada sesión de pruebas. </li><li> El dolor de pruebas de hipersensibilidad se debe realizar varias veces antes de ICC (basal) y en diferentes puntos temporales después de ICC, según se desee. </li></ol> 3. Umbral de retirada mecánica <ol><li> En el día de la prueba, las ratas lugar en la jaula con piso de malla de pruebas elevada 15-30 minutos antes de medir los umbrales de retirada, permitiendo jaula exploración y Grooming actividades de cese y las ratas se vuelvan más frágiles y estacionarios. </li><li> Utilizando una dinámica plantar von Frey aesthesiometer (Ugo Basile, Italia), la posición de la unidad de estimulador táctil con el filamento directamente debajo de la superficie a mediados de plantar de la pata trasera, evitando la malla de metal, y pulse la tecla de inicio. Esto levanta el filamento para estimular mecánicamente la superficie plantar de la pata trasera con una fuerza cada vez mayor. </li></ol> Manual von Frey pelos son una serie de filamentos de diámetro creciente que están calibradas para inducir una fuerza específica cuando se aplica a la piel. Se pueden utilizar como una alternativa a una electrónica dinámica plantar aesthesiometer von Frey para la medición de umbral retirada de la pata, como se ha descrito previamente 4. También hay varios fabricantes alternativos de electrónica de von Frey aesthesiometers (por ejemplo, CITI, EE.UU. y Somedic, Suecia). <ol start="3"><li> Registre el umbral de retirada mecánica de hindpa tanto lesionados y no lesionadosws, en un orden alternado. Este dispositivo se aplica presión a la pata y sobre la reacción de abstinencia, el sistema automáticamente registra y muestra la fuerza máxima aplicada (en gramos). Si una respuesta ambigua se produce (es decir, en el caso de deambulación o aseo), repetir el estímulo y registrar el umbral de nuevo. Lamer o temblor de la pata trasera después de la estimulación es un signo de comportamiento asociado con el dolor. </li><li> Repita el estímulo mecánico 3 veces con un intervalo de unos 5 minutos entre los estímulos, los umbrales de grabar, y luego calcular la media de los umbrales de retirada de la pata. </li></ol> 4. Latencia de retirada térmica El infrarrojo (IR) la intensidad del calor de la prueba plantar instrumento debe ser ajustado antes de iniciar el experimento. En nuestro laboratorio, ajustar la intensidad de IR para dar una latencia de retirada de la pata media de unos 10 segundos en ratas ingenuas, y nos pusimos en la latencia de corte de 20 segundos para evitar daños del tejidoedad en caso de no retirar una pata. <ol><li> En el día de la prueba, las ratas lugar en la jaula de vidrio pruebas piso de 15-30 minutos antes de medir la latencia, permitiendo exploración jaula y acicalamiento actividades de cesar y ratas para convertirse relajado y estacionario. </li><li> El uso de un instrumento de la analgesia térmica plantar (Ugo Basile, Italia), la posición de la fuente de luz infrarroja directamente debajo de la superficie a mediados de plantar de la pata trasera, y pulse la tecla de inicio. Esto expone la superficie plantar de la pata trasera de un haz de calor radiante a través de una superficie de vidrio transparente. También hay fabricantes alternativos de instrumentos térmicos analgesia plantares (por ejemplo, CITI, EE.UU. y Stoelting, EE.UU.). </li><li> Registre la latencia de retirada de las patas traseras, tanto lesionados y no lesionados, en un orden alternado. Este dispositivo registra automáticamente el tiempo necesario (en segundos) desde el inicio del estímulo térmico para retirada de la pata de la fuente de calor. Si una respuesta ambigua se produce (es decir, en el caso de deambulaciónción o de aseo), repita el estímulo y registrar la latencia de nuevo. Lamer o temblor de la pata trasera después de la estimulación es un signo de comportamiento asociado con el dolor. </li><li> Repita el estímulo de calor por lo menos 3 veces con un intervalo de unos 5 minutos entre los estímulos, las latencias de grabar, y luego calcular la media de las latencias de retirada de la pata. </li></ol> 4. Los resultados representativos Un ejemplo de umbral de retirada mecánica y térmica de latencia de retirada en las patas ipsilaterales (lesionado) y contralateral (sano) de ratas Wistar se muestra antes (basal) y 2-12 días después de la CCI en la figura 3A, y la Figura 3B. A partir de estas figuras puede verse que CCI provoca una reducción significativa en ambos umbrales de retirada mecánicas y térmicas latencias de retirada de la pata trasera ipsolateral, indicando la presencia de hipersensibilidad dolor mecánico y térmico, ambas características del dolor crónico neuropático<html> . <img alt="Figura 1" src="/files/ftp_upload/3393/3393fig1.jpg" /> Figura 1. Un esquemático que ilustra el posicionamiento de las ligaduras en el modelo de CCI. Este modelo animal de dolor neuropático implica la colocación de cuatro sueltas del intestino crómico-ligaduras en el nervio ciático, a nivel de la mitad del muslo. Es el resultado de que sólo daña una porción de las fibras aferentes que van a la hipersensibilidad dolor de pie y subsiguientes de la pata trasera. <img alt="Figura 2" src="/files/ftp_upload/3393/3393fig2.jpg" /> Figura 2. Diagrama de la zona de pata trasera utilizado para la prueba de dolor. La superficie a mediados de plantar justo por detrás de las almohadillas, como se indica por el círculo de puntos de color rojo, se utiliza para la prueba de dolor. Esta área está inervada principalmente por las ramas terminales del nervio ciático. El talón y las almohadillas se deben evitar debido a la sensibilidad diferencial de la piel. 3.jpg &quot;/&gt; Figura 3. El dolor de hipersensibilidad después de la constricción crónica del nervio ciático en ratas. Después de ICC, hay una reducción significativa en el umbral de retirada de la pata a estímulos mecánicos (A) y la latencia retirada de la pata a los estímulos térmicos (B) en el lesionado (ipsilateral) en comparación con la pata trasera no lesionado (contralateral) pata trasera (P &lt;0,01 medidas repetidas 2 -way ANOVA). Los datos muestran la media ± SEM, n = 7 ratas 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>

Los experimentos con animales fueron aprobados por el Cuidado de Animales y el Comité de Ética de la Universidad de New South Wales, Australia, y siguieron las directrices emitidas por la Asociación Internacional para el Estudio del Dolor.

CCI es un modelo ampliamente utilizado lesión del nervio periférico del dolor neuropático crónico. Es relativamente fácil de realizar, y produce hipersensibilidad dolor robusta y estable durante al menos un mes después de la lesión. Después de CCI, las ratas muestran una postura anormal de la pata trasera heridos (dedos de los pies unidos y flexión plantar y eversión de la pata), así como repetidos temblores, vigilancia y lamido de la pata trasera lesionado sugiere la presencia de dolor espontáneo 9.</s...

<table border="1"><tbody><tr><td> Nombre del reactivo / equipo </td><td> Empresa </td><td> Número de catálogo </td><td> Comentarios </td></tr><tr><td> Crómico intestino </td><td> Ethicon </td><td> T-211 </td><td> Espesor de 4-0 </td></tr><tr><td> El isoflurano </td><td> Delvet Pty. Ltd., Seven Hills, NSW </td><td> n / a </td><td></td></tr><tr><td> Mersilk </td><td> Ethicon </td><td> W-580 </td><td> 5-0 de espesor </td></tr><tr><td> Autoclip </td><td> Becton Dickinson </td><td> 427631 </td><td> 9 mm de acero inoxidable </td></tr><tr><td> Riodine </td><td> Orión </td><td> R1000802F </td><td> 1% w / v yodo </td></tr><tr><td> Instrumento de la analgesia térmica plantar </td><td> Ugo Basile </td><td> 37370 </td><td></td></tr><tr><td> Aesthesiometer plantar dinámica </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

El dolor neuropático crónico, como resultado del daño al sistema nervioso central o periférico, es una enfermedad prevalente y debilitante, que afecta a 18.7% de la población de 1,2. Los síntomas incluyen espontánea (hormigueo, ardor, una descarga eléctrica al igual que) el dolor, disestesias, parestesias, alodinia (dolor debido a que normalmente no dolorosos estímulos) y la hiperalgesia (una mayor respuesta a estímulos dolorosos). Los síntomas sensoriales son co-mórbida con discapacidades de comportamiento, tales como el insomnio y la depresión. Para estudiar el dolor neuropático crónico varios modelos animales que simulan lesiones de nervios periféricos han sido desarrollados, uno de los más utilizados es Bennett y (1988) lesión unilateral del nervio ciático Xie constricción crónica (ICC) 3 (Figura 1). Aquí presentamos un método para realizar pruebas de hipersensibilidad CCI y el dolor. ICC se realiza bajo anestesia, con el nervio ciático, por un lado expuesto por decisiones comoincisión de los familiares, y el corte a través del tejido conectivo entre los glúteos y los músculos superficial de bíceps femoral. Cuatro ligaduras catgut crómico se atan libremente alrededor del nervio ciático a intervalos de 1 mm, para ocluir solo, pero no detener el flujo de sangre epineural. La herida se cierra con suturas en el músculo y grapas en la piel. El animal se deja entonces recuperarse de la cirugía durante 24 horas antes de la prueba dolor hipersensibilidad comienza. Para las pruebas de comportamiento, las ratas se colocan en el aparato de ensayo y se les permite habituar al procedimiento de prueba. El área de pruebas es la mitad de la superficie plantar de la pata trasera (Figura 2), que cae dentro de la distribución del nervio ciático. Umbral de retirada mecánica se evaluó mediante la estimulación mecánica ambas patas traseras lesionados y no lesionados con una dinámica electrónica plantar von Frey aesthesiometer o manual de von Frey pelos 4. El umbral de retirada mecánica es la presión máxima ejercida(En gramos) que desencadena la retirada de la pata. Para la medición de latencia térmica retirada, primero descrito por Hargreaves et al (1988), la pata trasera está expuesto a un haz de calor radiante a través de una superficie de vidrio transparente con un plantar metros analgesia 5,6. La latencia de retirada de los estímulos de calor se registra como el tiempo de retirada de la pata en las dos patas traseras lesionados y no lesionados. Tras ICC, umbral de retirada mecánica, así como la retirada de latencia térmica en la pata lesionada son significativamente reducido, en comparación con las mediciones de referencia y la pata no lesionado (Figura 3). El ICC modelo de lesión del nervio periférico junto con las pruebas de hipersensibilidad dolor proporciona un sistema modelo para investigar la eficacia de los agentes terapéuticos potenciales para modificar el dolor neuropático crónico. En nuestro laboratorio, utilizamos ICC junto con la sensibilidad térmica y mecánica de las patas traseras para investigar el papel de las interacciones neuro-inmune en la patogénesisy el tratamiento del dolor neuropático.

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

Debido a la simplicidad de la cirugía y el resultado del comportamiento robusto, constricción crónica del nervio ciático es uno de los modelos animales de pre-eminentes de dolor neuropático. Dentro de las 24 horas siguientes a la cirugía, la hipersensibilidad del dolor se establece y se puede medir cuantitativamente utilizando una de von Frey aesthesiometer (ensayos mecánicos) y el medidor de la analgesia plantar (ensayo térmico).

La constricción crónica del nervio ciático y pruebas de hipersensibilidad dolor en ratas

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

58.5K vistas.  University of New South Wales. Debido a la simplicidad de la cirugía y el resultado del comportamiento robusto, constricción crónica del nervio ciático es uno de los modelos animales de pre-eminentes de dolor neuropático. Dentro de las 24 horas siguientes a la cirugía, la hipersensibilidad del dolor se establece y se puede medir cuantitativamente utilizando una de von Frey aesthesiometer (ensayos mecánicos) y el medidor de la analgesia plantar (ensayo térmico).

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

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 y crónica Prueba sensorial táctil después de la lesión de la médula espinal en ratas

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

Investigación

Medicina

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.

Las respuestas de la Vejiga Urinaria Distensión Evocados visceromotor como modelo para la vejiga Dolor en ratones

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.

El nervio ciático esposar modelo de dolor neuropático en ratones

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

Chronic Constriction Injury of the Rat's Infraorbital Nerve (IoN-CCI) to Study Trigeminal Neuropathic Pain

Animal models are important tools to study the pathophysiology and pharmacology of neuropathic pain. This manuscript describes the surgical and behavioral procedures to study trigeminal neuropathic pain in rats. To meet the specificity of trigeminal neuropathic pain syndromes, the infraorbital nerve (IoN) is subjected to a chronic constriction injury (CCI) by loosely ligating the nerve. An intra-orbital approach is presented here to expose and ligate the IoN in the orbital cavity. After IoN ligation, rats exhibit changes in spontaneous behavior and in response to von Frey hair stimulation that are indicative of persistent pain and mechanical allodynia. Two phases can be defined in the development of the behavioral changes. During the first week following IoN-CCI (phase 1), rats show an increased and asymmetric face grooming activity, i.e., with face wash strokes primarily directed to the nerve-injured IoN territory. A distinction is made between face grooming behavior that is part of a more general body grooming behavior, which remains largely unaffected by IoN-CCI, and face grooming that is neither preceded nor followed by body grooming, which is significantly increased after IoN-CCI. During this period, responsiveness to mechanical stimulation of the IoN territory is reduced. This hyporesponsiveness is abruptly replaced by an extreme hyperresponsiveness whereby even very weak stimulus intensities provoke nocifensive behavior (phase 2). The phenomenological similarities between these behavioral alterations and reported signs of facial pain (i.e., responses to noxious stimulation of the face) suggest the presence of dysesthesia/paresthesia and mechanical allodynia in the ligated IoN territory.

Chronic Constriction Injury of the Rat&#039;s Infraorbital Nerve IoN-CCI to Study Trigeminal Neuropathic Pain

This manuscript describes a rodent model to study trigeminal neuropathic pain. The methods described include surgical procedures to perform a chronic constriction injury of the rat&#8217;s infraorbital nerve and the post-surgical behavioral tests to measure the changes in spontaneous and evoked behavior that are indicative of persistent pain and allodynia.

Lesión crónica constricción del nervio infraorbitario de la Rata (Ion-CCI) para el Estudio del trigémino Dolor Neuropático

Animal models are important tools to study the pathophysiology and pharmacology of neuropathic pain. This manuscript describes the surgical and behavioral ...

A Model of Cardiac Remodeling Through Constriction of the Abdominal Aorta in Rats

Heart failure is one of the leading causes of death worldwide. It is a complex clinical syndromethat includes fatigue, dyspnea, exercise intolerance, and fluid retention. Changes in myocardial structure, electrical conduction, and energy metabolism develop with heart failure, leading to contractile dysfunction, increased risk of arrhythmias, and sudden death. Hypertensive heart disease is one of the key contributing factors of cardiac remodeling associated with heart failure. The most commonly-used animal model mimicking hypertensive heart disease is created via surgical interventions, such as by narrowing the aorta. Abdominal aortic constriction is a useful experimental technique to induce a pressure overload, which leads to heart failure. The surgery can be easily performed, without the need for chest opening or mechanical ventilation. Abdominal aortic constriction-induced cardiac pathology progresses gradually, making this model relevant to clinical hypertensive heart failure. Cardiac injury and remodeling can be observed 10 weeks after the surgery. The method described here provides a simple and effective approach to produce a hypertensive heart disease animal model that is suitable for studying disease mechanisms and for testing novel therapeutics.

A rat model of abdominal aortic constriction that induces cardiac hypertrophy and remodeling is described. An efficient, highly-reproducible, and minimally-invasive method is used to provide a simple yet useful platform for research in myocardial hypertrophy and dysfunction.

Un modelo de remodelamiento cardiaco A través de la constricción de la aorta abdominal en ratas

Heart failure is one of the leading causes of death worldwide. It is a complex clinical syndromethat includes fatigue, dyspnea, exercise intolerance, and ...

Minimally Invasive Transverse Aortic Constriction in Mice

Minimally invasive transverse aortic constriction (MTAC) is a more desirable method for the constriction of the transverse aorta in mice than standard open-chest transverse aortic constriction (TAC). Although transverse aortic constriction is a highly functional method for the induction of high pressure in the left ventricle, it is a more difficult and lengthy procedure due to its use of artificial ventilation with tracheal intubation. TAC is oftentimes also less survivable, as the newer method, MTAC, neither requires the cutting of the ribs and intercostal muscles nor tracheal intubation with a ventilation setup. In MTAC, as opposed to a thoracotomy to access to the chest cavity, the aortic arch is reached through a midline incision in the anterior neck. The thyroid is pulled back to reveal the sternal notch. The sternum is subsequently cut down to the second rib level, and the aortic arch is reached simply by separating the connective tissues and thymus. From there, a suture can be wrapped around the arch and tied with a spacer, and then the sternal cut and skin can be closed. MTAC is a much faster and less invasive way to induce left ventricular hypertension and enables the possibility for high-throughput studies. The success of the constriction can be verified using high-frequency trans-thoracic echocardiography, particularly color Doppler and pulsed-wave Doppler, to determine the flow velocities of the aortic arch and left and right carotid arteries, the dimension of the blood vessels, and the left ventricular function and morphology. A successful constriction will also trigger significant histopathological changes, such as cardiac muscle cell hypertrophy with interstitial and perivascular fibrosis. Here, the procedure of MTAC is described, demonstrating how the resulting flow changes in the carotid arteries can be examined with echocardiography, gross morphology, and histopathological changes in the heart.

Minimally invasive transverse aortic constriction (MTAC) conserves the essentials of regular transverse aortic constriction (TAC) while eliminating the use of a ventilator with tracheal intubation. It proves to be a highly desirable method for high-throughput studies on left ventricular overload, particularly in translational studies.

La constricción aórtica mínimamente invasiva transversal en ratones

Minimally invasive transverse aortic constriction (MTAC) is a more desirable method for the constriction of the transverse aorta in mice than standard ...

Development of Recombinant Proteins to Treat Chronic Pain

Chronic pain is difficult to treat and new approaches to resolve persistent pain are urgently needed. Anti-inflammatory cytokines are promising candidates for treating debilitating pain conditions due to their capacity to regulate aberrant neuro-immune interactions. However, physiologically they work in a network of various cytokines, and therefore their therapeutic effect may not be optimal when used as stand-alone drugs. To overcome this limitation, we developed a fusion protein of the anti-inflammatory cytokines IL4 and IL10. Here, we describe the methods for production and quality control of IL4-10 recombinant fusion protein and we test the effectiveness of the IL4-10 fusion protein to resolve pain in a mouse model of persistent inflammatory pain.

In this paper we provide details for the methods of production and quality control of an IL4-10 recombinant fusion protein. We also show how to test the effectiveness of this protein to resolve pain in a mouse model of inflammatory pain.

Desarrollo de proteínas recombinantes para el tratamiento de dolor crónico

Chronic pain is difficult to treat and new approaches to resolve persistent pain are urgently needed. Anti-inflammatory cytokines are promising candidates ...

Osteoarthritis Pain Model Induced by Intra-Articular Injection of Mono-Iodoacetate in Rats

The current animal models of osteoarthritis (OA) can be divided into spontaneous models and induced models, both of which aim to simulate the pathophysiological changes of human OA. However, as the main symptom in the late stage of OA, pain affects the patients&#39; daily life, and there are not many available models. The mono-iodoacetate (MIA)-induced model is the most widely used OA pain model, mainly used in rodents. MIA is an inhibitor of glyceraldehyde-3-phosphate dehydrogenase, which causes chondrocyte death, cartilage degeneration, osteophyte, and measurable changes in animal behavior. Besides, expression changes of matrix metalloproteinase (MMP) and pro-inflammatory cytokines (IL1 &#946; and TNF &#945;) can be detected in the MIA-induced model. Those changes are consistent with OA pathophysiological conditions in humans, indicating that MIA can induce a measurable and successful OA pain model. This study aims to describe the methodology of intra-articular injection of MIA in rats and discuss the resulting pain-related behaviors and histopathological changes.

This study describes the method of intra-articular injection of mono-iodoacetate in rats and discusses the resulting pain-related behaviors and histopathological changes, which provide references for future applications.

Modelo de dolor de osteoartritis inducido por inyección intraarticular de monoyodoacetato en ratas

The current animal models of osteoarthritis (OA) can be divided into spontaneous models and induced models, both of which aim to simulate the ...

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 del nervio ciático: un modelo de ratón de dolor neuropático crónico para estudiar el efecto antinociceptivo de nuevas terapias

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

Mejora de la terapia con FSN en el modelo de rata: minimización de las molestias y maximización de la eficiencia

Efficacy of Fu's Subcutaneous Needling on Sciatic Nerve Pain: Behavioral and Electrophysiological Changes in a Chronic Constriction Injury Rat Model

Fu&#39;s subcutaneous needling (FSN), an invented acupuncture technique from traditional Chinese medicine, is used worldwide for pain relief. However, the mechanisms of action are still not fully understood. During FSN treatment, the FSN needle is inserted and retained in the subcutaneous tissues for a long duration with a swaying movement. However, challenges arise from maintaining a posture while manipulating FSN in animal models (e.g., rats) for researchers. Uncomfortable treatment can lead to fear and resistance to FSN needles, increasing the risk of injury and may even affect research data. Anesthesia may also affect the study results too. Hence, there is a need for strategies in FSN therapy on animals that minimize injury during the intervention. This study employs a chronic constriction injury model in Sprague-Dawley rats to induce neuropathic pain. This model replicates the pain induced by nerve injury observed in humans through surgical constriction of a peripheral nerve, mimicking the compression or entrapment seen in conditions such as nerve compression syndromes and peripheral neuropathies. We introduce an appropriate manipulation for easily inserting an FSN needle into the subcutaneous layer of the animal&#39;s body, including needle insertion and direction, needle retention, and swaying movement. Minimizing the rat&#39;s discomfort prevents the rat from being tense, which causes the muscle to contract and hinder the entry of the needle and improves the study efficiency.

Autor destacado: Desvelando los efectos terapéuticos del tratamiento con FSN: uniendo la investigación y las aplicaciones clínicas en el dolor neuropático

We present a protocol for using Fu's subcutaneous needling in a chronic constriction injury model to induce sciatic nerve pain in rats.

Eficacia de la punción subcutánea de Fu en el dolor del nervio ciático: cambios conductuales y electrofisiológicos en un modelo de rata con lesión por constricción crónica

Fu&#39;s subcutaneous needling (FSN), an invented acupuncture technique from traditional Chinese medicine, is used worldwide for pain relief. However, the ...