Name: a mouse model of fatigue induced by peripheral irradiation
Uploaded: 2017-03-17T14:00:00.000+00:00
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Description: Watch this Scientific Journal Video about A Mouse Model of Fatigue Induced by Peripheral Irradiation at JoVE.com

Los autores desean agradecer a Michele Allen del National Heart, Lung, and Blood Institute (NHLBI) de los Institutos Nacionales de Salud (NIH) por compartir generosamente su experiencia en métodos de fenotipado murinos y por su asistencia técnica en curso, así como para timothy Hunt, de NHLBI para ayudarnos a desarrollar el dispositivo de protección. Este estudio es apoyado por la División de Investigación Intramural del Instituto Nacional de Investigación en Enfermería del NIH, y parte de la prueba de validación es apoyado por una beca de la Oncology Nursing Society Foundation.

Declaración de Ética: Este estudio fue aprobado por los Institutos Nacionales de Salud (NIH) Cuidado de Animales y el empleo Comisión. Todos los investigadores que participan en el manejo de animales y medición de los resultados del estudio fueron debidamente capacitados por la Oficina de NIH de Cuidado y Uso de Animales y el Instituto Nacional del Corazón, Pulmón y la Sangre murino Fenotipificación Core. Todos los aspectos de la experimentación con animales, la vivienda y las condiciones ambientales utilizados en este estudio fueron en cumplimiento de la Guía para el Cuidado y Uso de Animales de Laboratorio 11. 1. Vivienda y los animales de experimentación NOTA: Casa de sexo masculino ratones C57BL / 6 (aproximadamente cinco semanas de edad a la llegada) de forma individual durante todo el experimento y proporcionar acceso ad libitum a comida y agua. Todas las jaulas se mantienen en un ciclo de luz-oscuridad de 12:12 h con la fase de la luz a partir de las 6 am y fase oscura a las 6 pm. <ol><li> Identificar los ratones y asignarlos a ventilar norma individuald jaulas de ratón. Espere 24 horas después de que el procedimiento de identificación para la recuperación. NOTA: Tatuaje de la cola se recomienda como un medio de identificación para eliminar la posibilidad de que una etiqueta de oreja puede quedar atrapada en una rueda para correr. Tatuar un número en la cola de cada ratón, con el número en la cola coincide con el número escrito en la jaula del ratón. </li><li> Permitir que los ratones se aclimaten a sus jaulas durante al menos tres días más, el manejo de cada ratón suavemente durante un período de tres minutos por día. </li></ol> 2. rueda corriente Aclimatación y Línea de Base <ol><li> Introducir a los ratones a jaulas individuales VWRA, cada uno equipado con una rueda para correr conectada a un contador electrónico para el registro continuo. NOTA: Todos los contadores de ruedas se conectan a un ordenador a través de una única interfaz USB (consulte la Lista de Materiales). El software del ordenador calcula el número de rotaciones de la rueda, la distancia recorrida, y la velocidad media a través de cada intervalo de tiempo designado de la duratio total especificadonorte. Una vez que la grabación se detiene, los datos se guardan automáticamente tanto como texto y hojas de cálculo. </li><li> Iniciar la grabación de VWRA a través de la interfaz de programas de computación. Establecer intervalos de grabación a una hora y la duración de al menos cinco días. VWRA seguir grabando durante al menos cinco días. NOTA: Al final de la etapa 2.2, todos los ratones deben lograr una cantidad relativamente constante de la actividad diaria de la rueda en marcha. Si no es así, identificar y excluir cualquier valor atípico. </li><li> Detener VWRA grabación a través de la interfaz del software, y volver a los ratones a sus jaulas estándar descritas en el paso 1.1 (jaulas sin ruedas corriendo). </li><li> Selección aleatoria de los ratones en cualquiera de los controles irradiados o grupos irradiados. </li></ol> 3. La irradiación Nota: Realice los siguientes pasos para todos los ratones en ambos grupos, una vez al día durante tres días consecutivos. Tratar los ratones en el mismo orden cada día. <ol><li> Anestesiar a cada ratón mediante inyección intraperitoneal deuna mezcla de ketamina (100 mg / kg) y xilazina (10 mg / kg) mezcla. </li><li> Confirmar la anestesia con una pizca dedo del pie y el uso de pomada en los ojos para evitar la sequedad mientras que bajo anestesia. </li><li> Transferir el ratón anestesiado en un dispositivo de protección de plomo. Organizar el ratón en el blindaje de modo que sólo se expone la región pélvica / abdominal inferior. NOTA: El blindaje se compone de dos de plomo &quot;cajas&quot;, con un espacio abierto entre estrecha en que permite la exposición de radiación a una pequeña región, dirigida del ratón (véase la Figura 1). </li><li> Use cinta adhesiva médica para asegurar la base de la cola del ratón en posición dentro del blindaje. NOTA: El paso 3.4 es opcional, pero al hacerlo puede ayudar a asegurar que la posición del ratón no cambia durante el siguiente paso. </li><li> Transporte el dispositivo de protección en el irradiador, asegurándose de que la posición de los animales en el blindaje se mantiene. </li><li> Si el ratón está en el grupo de irradiación, entregar 800 cGy a dostasa de e de aproximadamente 110 cGy / min. Si el ratón está en el grupo control simulado de la irradiación, deje el puntero del ratón en el irradiador inactivo durante el tiempo equivalente. Nota: Los valores irradiator óptimas dependerán del dispositivo en particular. La tasa de dosis de 110 cGy / min suministrado desde una fuente de cesio 137 es la tasa de dosis central del irradiador utilizado aquí. El tiempo de exposición se ajustó para alcanzar la dosis total deseada de 800 cGy. </li><li> Retire el ratón del irradiador y el blindaje, y luego devolverlo a su estado original jaula, las normas mencionadas en el paso 1.1. </li><li> Monitorear continuamente el ratón hasta que se haya recuperado el conocimiento suficiente para mantener decúbito esternal. </li></ol> 4. Medición de fatiga inducida por radiación <ol><li> En el día después de la finalización de tres días sucesivos de la irradiación, la transferencia de los ratones a sus jaulas VWRA individuales descritos en el paso 2.1. </li><li> Record VWRA, como se describe en el paso 2.2, con la excepción aquí de ajustar la grabaciónduración de más de 15 días. Al final de los 15 días, detener manualmente VWRA grabación a través de la interfaz de software. NOTA: Los datos de cada período de registro se guardan automáticamente como hojas de cálculo, cada uno de los cuales incluye la rotación, la distancia y las mediciones de velocidad para todos los animales (columnas) y en todos los intervalos (filas) a lo largo de la duración de la grabación. Al final del experimento, hay dos hojas de cálculo generadas por el software de grabación: una para el VWRA pre-irradiación, y uno para el VWRA post-irradiación. </li></ol>

<ol>
	<li>Minton, O., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cancer-related+fatigue+and+its+impact+on+functioning.">Cancer-related fatigue and its impact on functioning.</a> Cancer. 119, 2124-2130 (2013).</li><li>Bower, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cancer-related+fatigue--mechanisms,+risk+factors,+and+treatments.">Cancer-related fatigue--mechanisms, risk factors, and treatments.</a> Nat Rev Clin Oncol. 11 (10), 597-609 (2014).</li><li>Saligan, L. N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+biology+of+cancer-related+fatigue:+a+review+of+the+literature.">The biology of cancer-related fatigue: a review of the literature.</a> Support Care Cancer. 23 (8), 2461-2478 (2015).</li><li>Norden, D. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tumor+growth+increases+neuroinflammation,+fatigue+and+depressive-like+behavior+prior+to+alterations+in+muscle+function.">Tumor growth increases neuroinflammation, fatigue and depressive-like behavior prior to alterations in muscle function.</a> Brain Behav Immun. 43, 76-85 (2015).</li><li>Ray, M. A., Trammell, R. A., Verhulst, S., Ran, S., Toth, L. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+a+mouse+model+for+assessing+fatigue+during+chemotherapy.">Development of a mouse model for assessing fatigue during chemotherapy.</a> Comp Med. 61 (2), 119-130 (2011).</li><li>Zombeck, J. A., Fey, E. G., Lyng, G. D., Sonis, S. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+clinically+translatable+mouse+model+for+chemotherapy-related+fatigue.">A clinically translatable mouse model for chemotherapy-related fatigue.</a> Comp Med. 63 (6), 491-497 (2013).</li><li>Renner, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+murine+model+of+peripheral+irradiation-induced+fatigue.">A murine model of peripheral irradiation-induced fatigue.</a> Behav Brain Res. 307, 218-226 (2016).</li><li>Van der Meeren, A., Lebaron-Jacobs, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioural+consequences+of+an+8+Gy+total+body+irradiation+in+mice:+Regulation+by+interleukin-4.">Behavioural consequences of an 8 Gy total body irradiation in mice: Regulation by interleukin-4.</a> Canadian Journal of Physiology and Pharmacology. 79 (2), 140-143 (2001).</li><li>York, J. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+biobehavioral+and+neuroimmune+impact+of+low-dose+ionizing+radiation.">The biobehavioral and neuroimmune impact of low-dose ionizing radiation.</a> Brain Behav Immun. 26 (2), 218-227 (2012).</li><li>Meijer, J. H., Robbers, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wheel+running+in+the+wild.">Wheel running in the wild.</a> Proc Biol Sci. 281 (1786), (2014).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The Guide for the Care and Use of Laboratory Animals. , (2011).</li><li>Heredia, L., Torrente, M., Domingo, J. L., Colomina, M. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Individual+housing+and+handling+procedures+modify+anxiety+levels+of+Tg2576+mice+assessed+in+the+zero+maze+test.">Individual housing and handling procedures modify anxiety levels of Tg2576 mice assessed in the zero maze test.</a> Physiol Behav. 107 (2), 187-191 (2012).</li><li>Varty, G. B., Paulus, M. P., Braff, D. L., Geyer, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Environmental+enrichment+and+isolation+rearing+in+the+rat:+effects+on+locomotor+behavior+and+startle+response+plasticity.">Environmental enrichment and isolation rearing in the rat: effects on locomotor behavior and startle response plasticity.</a> Biol Psychiatry. 47 (10), 864-873 (2000).</li><li>Pham, T. M., Brene, S., Baumans, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+assessment+of+intermittent+wheel+running+and+individual+housing+in+mice+in+the+laboratory.">Behavioral assessment of intermittent wheel running and individual housing in mice in the laboratory.</a> J Appl Anim Welf Sci. 8 (3), 157-173 (2005).</li><li>Knab, A. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Repeatability+of+exercise+behaviors+in+mice.">Repeatability of exercise behaviors in mice.</a> Physiol Behav. 98 (4), 433-440 (2009).</li><li>Novak, C. M., Burghardt, P. R., Levine, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+a+running+wheel+to+measure+activity+in+rodents:+relationship+to+energy+balance,+general+activity,+and+reward.">The use of a running wheel to measure activity in rodents: relationship to energy balance, general activity, and reward.</a> Neurosci Biobehav Rev. 36 (3), 1001-1014 (2012).</li><li>Mineur, Y. S., Belzung, C., Crusio, W. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+unpredictable+chronic+mild+stress+on+anxiety+and+depression-like+behavior+in+mice.">Effects of unpredictable chronic mild stress on anxiety and depression-like behavior in mice.</a> Behav Brain Res. 175 (1), 43-50 (2006).</li><li>Perhach, J. L., Barry, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stress+responses+of+rats+to+acute+body+or+neck+restraint.">Stress responses of rats to acute body or neck restraint.</a> Physiol Behav. 5 (4), 443-448 (1970).</li><li>Iwakawa, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Different+radiation+susceptibility+among+five+strains+of+mice+detected+by+a+skin+reaction.">Different radiation susceptibility among five strains of mice detected by a skin reaction.</a> J Radiat Res. 44 (1), 7-13 (2003).</li><li>Travis, E. L., Peters, L. J., McNeill, J., Thames, H. D., Karolis, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+dose-rate+on+total+body+irradiation:+lethality+and+pathologic+findings.">Effect of dose-rate on total body irradiation: lethality and pathologic findings.</a> Radiother Oncol. 4 (4), 341-351 (1985).</li><li>Duran-Struuck, R., Dysko, R. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Principles+of+bone+marrow+transplantation+(BMT):+providing+optimal+veterinary+and+husbandry+care+to+irradiated+mice+in+BMT+studies.">Principles of bone marrow transplantation (BMT): providing optimal veterinary and husbandry care to irradiated mice in BMT studies.</a> J Am Assoc Lab Anim Sci. 48 (1), 11-22 (2009).</li><li>Duran-Struuck, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+susceptibility+of+C57BL/6NCr+and+B6.Cg-Ptprca+mice+to+commensal+bacteria+after+whole+body+irradiation+in+translational+bone+marrow+transplant+studies.">Differential susceptibility of C57BL/6NCr and B6.Cg-Ptprca mice to commensal bacteria after whole body irradiation in translational bone marrow transplant studies.</a> J Transl Med. 6, 10 (2008).</li></ol>

Esta investigación fue apoyada por la División de Investigación Intramural del Instituto Nacional de Investigación en Enfermería, Institutos Nacionales de Salud.

Hemos descrito un protocolo mediante el uso de la irradiación periférica dirigida a inducir una reducción de VWRA en ratones sin confundir la morbilidad o la mortalidad. Es importante destacar que un dispositivo de pantalla simple permite la irradiación en este protocolo para dirigirse a una región deseada consistente, imitando los tratamientos de radiación recibidas por los pacientes con cáncer pélvico. En contraste con los modelos existentes que implican CRF cerebral o irradiación corporal total <sup class="x...

La fatiga relacionada con el cáncer (CRF) es una condición penosa y costosa que a menudo afecta a los pacientes que reciben tratamientos contra el cáncer 1. La fatiga es ni proporcional a la actividad reciente ni aliviado por el reposo, y se asocia con una amplia variedad de trastornos relacionados con el estado de ánimo, la motivación, la atención y la cognición 2. Las causas biológicas de CRF son desconocidos, aunque se ha demostrado en muchos casos que se correlaciona con los niveles de inflamación y citoquinas, también en algunos casos con los niveles de hemoglobina y la función de varios sistemas hormonales (ver SALIGAN et al. 3 para una revisión de biológica estudios de CRF). Los estudios controlados utilizando modelos animales son necesarios para comprender el comportamiento y la biología asociada a esta enfermedad compleja. Mientras que, el 6 de grasa 4 o 5 relacionada con la quimioterapia relacionada con el tumorigue se ha estudiado en modelos de roedores, la etiología de la IRC-puede ser un tratamiento específico. Para investigar CRF relacionados con la terapia de radiación, nuestro grupo ha desarrollado recientemente un modelo de ratón de la fatiga inducida por irradiación 7. En contraste con los modelos existentes que implican CRF cerebral o irradiación corporal total 8, 9, este modelo explora cómo un cambio en el comportamiento impulsado centralmente, como la fatiga, puede ser provocada por un procedimiento de irradiación dirigida periféricamente. El procedimiento aquí descrito está diseñado para modelar la radioterapia se administra a pacientes con cáncer pélvico, usando protección de plomo para apuntar la región pélvica / abdominal inferior con irradiación. Sin embargo, mediante la modificación de la protección de plomo o su colocación con respecto a los animales de experimentación, este procedimiento podría ser adaptado para modelar irradiación de otras partes del cuerpo. Voluntaria actividad rueda de funcionamiento (VWRA) se utiliza para medir la fatiga behavio similarr; porque es un comportamiento voluntario y normal 10, se debe permitir el uso simultáneo de otras pruebas de comportamiento y biológicos. Hemos encontrado que la irradiación periférica es suficiente para reducir VWRA en ratones sin causar morbilidad abierta 7. Los futuros experimentos con este modelo pueden ayudar a revelar efectos de la irradiación periférica en la señalización biológica inmune y otros, así como los cambios aguas abajo en el sistema nervioso central que puede producir déficits asociados con CRF.

<table><tbody><tr><td>C57BL/6 Mice</td><td>Charles River</td><td></td><td>Strain code 027 (http://www.criver.com/products-services/basic-research/find-a-model/c57bl-6n-mouse)</td></tr><tr><td>Ketamine HCl</td><td>Putney</td><td></td><td>100 mg/mL stock solution</td></tr><tr><td>Xylazine HCl</td><td>Lloyd Laboratories</td><td></td><td>100 mg/mL stock solution</td></tr><tr><td>Rodent Tattoo System</td><td>AIMS</td><td>ATS-3</td><td>http://animalid.com/lab-animal-identification-systems/ats-3-general-rodent-tattoo-system</td></tr><tr><td>Lead Shielding Apparatus</td><td>(custom made)</td><td></td><td>One-inch thick lead shielding arranged as two boxes with a one-inch thick gap between them for targeted irradiation</td></tr><tr><td>Plexiglass shielding container</td><td>(custom made)</td><td></td><td>Plexiglass container filled with styrofoam. Styrofoam cutouts hold the lead shielding in place.</td></tr><tr><td>GammaCell 40 Exactor</td><td>Best Theratronics</td><td></td><td>http://www.theratronics.ca/product_gamma40.html</td></tr><tr><td>RAD Disk Ultra</td><td>Best Theratronics</td><td></td><td>http://www.theratronics.ca/product_rad.html</td></tr><tr><td>Mouse Single Activity Wheel Chamber</td><td>Lafayette Instrument Company</td><td>#80820</td><td>http://www.lafayetteneuroscience.com/product_detail.asp?itemid=980</td></tr><tr><td>Activity Wheel Counter for Computer Monitoring</td><td>Lafayette Instrument Company</td><td>#86061</td><td>http://www.lafayetteneuroscience.com/product_detail.asp?itemid=1052</td></tr><tr><td>Modular Cable for Wheel Counters</td><td>Lafayette Instrument Company</td><td>#86051-7</td><td>http://www.lafayetteneuroscience.com/product_detail.asp?itemid=1046</td></tr><tr><td>USB Computer Interface for Activity Wheel Counters</td><td>Lafayette Instrument Company</td><td>#86056A</td><td>http://www.lafayetteneuroscience.com/product_detail.asp?itemid=1047</td></tr><tr><td>Activity Wheel Monitor Software</td><td>Lafayette Instrument Company</td><td>#86065</td><td>http://www.lafayetteneuroscience.com/product_detail.asp?itemid=1053</td></tr></tbody></table>

a mouse model of fatigue induced by peripheral irradiation

la fatiga relacionada con el cáncer (CRF) es una condición penosa y costosa que a menudo afecta a los pacientes que reciben tratamientos contra el cáncer, incluyendo la terapia de radiación. Aquí se describe un método que utiliza la irradiación periférica dirigida a inducir un comportamiento similar a la fatiga en ratones. Con blindaje apropiado, la irradiación se dirige a la región pélvica / abdominal inferior del ratón, evitando el cerebro, en un esfuerzo para modelar el tratamiento de radiación recibida por los individuos con cáncer pélvico. Entregamos una dosis de irradiación que es suficiente para inducir un comportamiento a la fatiga como en ratones, medida por la actividad voluntaria rueda de funcionamiento (VWRA), sin causar morbilidad obvia. Desde rueda para correr es un comportamiento normal, voluntaria en ratones, su uso debe tener poco efecto de confusión en otras pruebas de comportamiento o medidas biológicas. Por lo tanto, la rueda de funcionamiento se puede utilizar como una medida de resultado factible en la comprensión de las correlaciones conductuales y biológicos de la fatiga. CRF es una afección compleja con el co frecuentesmorbilidades, y probablemente tiene causas relacionadas tanto con el cáncer y sus distintos tratamientos. Los métodos descritos en este documento son útiles para investigar los cambios inducidos por la radiación que contribuyen al desarrollo de la IRC y, más en general, para explorar las redes biológicas que pueden explicar el desarrollo y la persistencia de un comportamiento periférica provocada, pero centralmente impulsado similar a la fatiga .

Tres lotes de ratones se realizaron mediante el protocolo descrito anteriormente. Hubo un total de 16 farsa y 20 irradiadas (2.400 cGy, 3 x 800 / día cGy) ratones. Después de tres días consecutivos de la irradiación, el grupo irradiado demostraron una reducción significativa en comparación con VWRA simulado (mixtas ANOVA de medidas repetidas: efecto principal del tratamiento de irradiación, F 1,13 = 19,233, p &lt;0,001). El efecto fue significativo durante los primeros ...

Watch this Scientific Journal Video about A Mouse Model of Fatigue Induced by Peripheral Irradiation at JoVE.com

A Mouse Model of Fatigue Induced by Peripheral Irradiation

Se describe un método que usa irradiación periférica dirigida a inducir un comportamiento a la fatiga como en ratones. La dosis de irradiación no letal seleccionado conduce a una reducción de una semana de duración de la actividad voluntaria rueda de funcionamiento.

Un modelo de ratón de la fatiga inducida por irradiación periférica

Cancer-related fatigue (CRF) is a distressing and costly condition that often affects patients receiving cancer treatments, including radiation therapy. ...

a-mouse-model-of-fatigue-induced-by-peripheral-irradiation

Universidad de Cartagena UDEC

Research

JoVE Journal

Cancer Research

7.8K vistas.  National Institutes of Health.  Se describe un método que usa irradiación periférica dirigida a inducir un comportamiento a la fatiga como en ratones. La dosis de irradiación no letal seleccionado conduce a una reducción de una semana de duración de la actividad voluntaria rueda de funcionamiento. 

Video: A Mouse Model of Fatigue Induced by Peripheral Irradiation

Fluorescent Orthotopic Mouse Model of Pancreatic Cancer

Pancreatic cancer remains one of the cancers for which survival has not improved substantially in the last few decades. Only 7% of diagnosed patients will survive longer than five years. In order to understand and mimic the microenvironment of pancreatic tumors, we utilized a murine orthotopic model of pancreatic cancer that allows non-invasive imaging of tumor progression in real time. Pancreatic cancer cells expressing green fluorescent protein (PANC-1 GFP) were suspended in basement membrane matrix, high concentration, (e.g., Matrigel HC) with serum-free media and then injected into the tail of the pancreas via laparotomy. The cell suspension in the high concentration basement membrane matrix becomes a gel-like substance once it reaches room temperature; therefore, it gels when it comes in contact with the pancreas, creating a seal at the injection site and preventing any cell leakage. Tumor growth and metastasis to other organs are monitored in live animals by using fluorescence. It is critical to use the appropriate filters for excitation and emission of GFP. The steps for the orthotopic implantation are detailed in this article so researchers can easily replicate the procedure in nude mice. The main steps of this protocol are preparation of the cell suspension, surgical implantation, and whole body fluorescent in vivo imaging. This orthotopic model is designed to investigate the efficacy of novel therapeutics on primary and metastatic tumors.

A procedure to implant green fluorescent protein-expressing pancreatic cancer cells (PANC-1 GFP) orthotopically into the pancreas of Balb-c Ola Hsd-Fox1nu mice to assess tumor progression and metastasis is presented here.

Fluorescente modelo de ratón ortotópico de cáncer de páncreas

Pancreatic cancer remains one of the cancers for which survival has not improved substantially in the last few decades. Only 7% of diagnosed patients will ...

Investigación

Investigación sobre el cáncer

Evaluating the Role of Mitochondrial Function in Cancer-related Fatigue

Fatigue is a common and debilitating condition that affects most cancer patients. To date, fatigue remains poorly characterized with no diagnostic test to objectively measure the severity of this condition. Here we describe an optimized method for assessing mitochondrial function of PBMCs collected from fatigued cancer patients. Using a compact extracellular flux system and sequential injection of respiratory inhibitors, we examined PBMC mitochondrial functional status by measuring basal mitochondrial respiration, spare respiratory capacity, and energy phenotype, which describes the preferred energy pathway to respond to stress. Fresh PBMCs are readily available in the clinical setting using standard phlebotomy. The entire assay described in this protocol can be completed in less than 4 hours without the involvement of complex biochemical techniques. Additionally, we describe a normalization method that is necessary for obtaining reproducible data. The simple procedure and normalization methods presented allow for repeated sample collection from the same patient and generation of reproducible data that can be compared between time points to evaluate potential treatment effects.

Our goal was to develop a practical protocol to evaluate mitochondrial dysfunction associated with fatigue in cancer patients. This innovative protocol is optimized for clinical use involving only standard phlebotomy and basic laboratory procedures.

Evaluar el papel de la función mitocondrial en la fatiga relacionada con el cáncer

Fatigue is a common and debilitating condition that affects most cancer patients. To date, fatigue remains poorly characterized with no diagnostic test to ...

The Treadmill Fatigue Test: A Simple, High-throughput Assay of Fatigue-like Behavior for the Mouse

Fatigue is a prominent symptom in many diseases and disorders and reduces quality of life for many people. The lack of clear pathogenesis and failure of current interventions to adequately treat fatigue in all patients leaves a need for new treatment options. Despite the therapeutic need and importance of preclinical research in helping identify promising novel treatments, few preclinical assays of fatigue are available. Moreover, the most common preclinical assay used to assess fatigue-like behavior, voluntary wheel running, is not suitable for use with some strains of mice, may not be sensitive to drugs that reduce fatigue, and has relatively low throughput. The current protocol describes a novel, non-voluntary preclinical assay of fatigue-like behavior, the treadmill fatigue test, and provides evidence of its efficacy in detecting fatigue-like behavior in mice treated with a chemotherapy drug known to cause fatigue in humans and fatigue-like behavior in animals. This assay may be a beneficial alternative to wheel running, as fatigue-like behavior and potential interventions can be assessed in a greater number of mice over a shorter time frame, thus permitting faster discovery of new therapeutic options.

Fatigue is a common, undertreated and frequently poorly-understood symptom in many diseases and disorders. New preclinical assays of fatigue may help to improve current understanding and future treatment of fatigue. To that end, the current protocol provides a novel means of measuring fatigue-like behavior in the mouse.

El Ensayo de fatiga rueda de ardilla: Un ensayo sencillo, de gran procesamiento de comportamiento a fatiga similar para el ratón

Fatigue is a prominent symptom in many diseases and disorders and reduces quality of life for many people. The lack of clear pathogenesis and failure of ...

Comportamiento

Functional Interrogation of Adult Hypothalamic Neurogenesis with Focal Radiological Inhibition

The functional characterization of adult-born neurons remains a significant challenge. Approaches to inhibit adult neurogenesis via invasive viral delivery or transgenic animals have potential confounds that make interpretation of results from these studies difficult. New radiological tools are emerging, however, that allow one to noninvasively investigate the function of select groups of adult-born neurons through accurate and precise anatomical targeting in small animals. Focal ionizing radiation inhibits the birth and differentiation of new neurons, and allows targeting of specific neural progenitor regions. In order to illuminate the potential functional role that adult hypothalamic neurogenesis plays in the regulation of physiological processes, we developed a noninvasive focal irradiation technique to selectively inhibit the birth of adult-born neurons in the hypothalamic median eminence. We describe a method for Computer tomography-guided focal irradiation (CFIR) delivery to enable precise and accurate anatomical targeting in small animals. CFIR uses three-dimensional volumetric image guidance for localization and targeting of the radiation dose, minimizes radiation exposure to nontargeted brain regions, and allows for conformal dose distribution with sharp beam boundaries. This protocol allows one to ask questions regarding the function of adult-born neurons, but also opens areas to questions in areas of radiobiology, tumor biology, and immunology. These radiological tools will facilitate the translation of discoveries at the bench to the bedside.

The function of adult-born mammalian neurons remains an active area of investigation. Ionizing radiation inhibits the birth of new neurons. Using computer tomography-guided focal irradiation (CFIR), three-dimensional anatomical targeting of specific neural progenitor populations can now be used to assess the functional role of adult neurogenesis.

Interrogatorio funcional de adultos hipotalámica neurogénesis con focal Radiológica Inhibición

The functional characterization of adult-born neurons remains a significant challenge. Approaches to inhibit adult neurogenesis via invasive viral ...

Neurociencias

Establishment of a Robust and Reproducible Model of Radiation-Induced Skin and Muscle Fibrosis

Radiation-induced skin fibrosis (RISF) can result from a plethora of scenarios including cancer therapy, accidental exposure, or acts of terrorism. Radioactive beams can penetrate through the skin and affect the structures in their path including skin, muscles, and internal organs. Skin is the first structure to get exposed to radiation and is susceptible to develop chronic fibrosis, which is challenging to treat. Currently, limited treatment options show moderate efficacy in mitigating radiation-related skin fibrosis. A key factor hindering the development of effective countermeasures is the absence of a convenient and robust model that could allow for translation of the experimental findings to humans. Here, a robust and reproducible murine hind limb skin fibrosis model has been established for prophylactic and therapeutic evaluation of possible agents for functional and molecular recovery.
The right hind limb was irradiated using a single dose of 40 (Gray) Gy to induce skin fibrosis. Subjects developed edema and dermatitis in the early stages proceeded by visible skin constriction. Irradiated limbs showed a significantly reduced limb range of motion in the following weeks. In late stages, acute side effects subsided, yet chronic fibrosis persisted. A gait index was performed as an additional functional assay, which demonstrated the development of functional impairment. These non-invasive methods demonstrated reliable measurements for tracing fibrosis progression, which is supported by histological analyses. The radiation dose, application, and post-irradiation analyses employed in this model offer a vigorous and reproducible method for studying radiation-induced skin fibrosis and testing the efficacy of therapeutical agents.

Here we present a protocol to induce radiation-induced skin fibrosis in the hind limb of mice and perform post-irradiation measurements of chronic impairment via limb excursion and gait index analyses to evaluate the functional outcome. The model elucidates radiation-related skin fibrosis mechanisms and is useful in subclinical therapeutic studies.

Radiation-induced skin fibrosis (RISF) can result from a plethora of scenarios including cancer therapy, accidental exposure, or acts of terrorism. ...

Diffuse Optical Spectroscopy for the Quantitative Assessment of Acute Ionizing Radiation Induced Skin Toxicity Using a Mouse Model

Acute skin toxicities from ionizing radiation (IR) are a common side effect from therapeutic courses of external beam radiation therapy (RT) and negatively impact patient quality of life and long term survival. Advances in the understanding of the biological pathways associated with normal tissue toxicities have allowed for the development of interventional drugs, however, current response studies are limited by a lack of quantitative metrics for assessing the severity of skin reactions. Here we present a diffuse optical spectroscopic (DOS) approach that provides quantitative optical biomarkers of skin response to radiation. We describe the instrumentation design of the DOS system as well as the inversion algorithm for extracting the optical parameters. Finally, to demonstrate clinical utility, we present representative data from a pre-clinical mouse model of radiation induced erythema and compare the results with a commonly employed visual scoring. The described DOS method offers an objective, high through-put evaluation of skin toxicity via functional response that is translatable to the clinical setting.

We present a diffuse optical spectroscopic (DOS) approach that provides quantitative optical biomarkers of skin response to radiation. We describe DOS instrumentation design, optical parameters extraction algorithms and the animal handling procedures required to yield representative data from a pre-clinical mouse model of radiation induced erythema.

Espectroscopia óptica difusa para la evaluación cuantitativa de la radiación ionizante aguda La toxicidad cutánea inducida utilizando un modelo de ratón

Acute skin toxicities from ionizing radiation (IR) are a common side effect from therapeutic courses of external beam radiation therapy (RT) and ...

Medicina

A Rat Model of Central Fatigue Using a Modified Multiple Platform Method

In this article, we introduced a rat model of central fatigue using the modified multiple platform method (MMPM). The Multiple Platform box was designed as a water tank with narrow platforms on the bottom. The model rats were put into the tank and stood on the platforms for 14 h (18:00 - 8:00) per day for a consecutive 21 days, with a blank control group set for contrast. At the end of modeling, rats in the model group showed an obvious fatigued appearance. To assess the model, we performed several behavioral tests, including the open field test (OFT), the elevated plus maze (EPM) test, and the exhaustive swimming (ES) test. The results showed that anxiety, spatial cognition impairment, poor muscle performance, and declined voluntary activity presented in model rats confirm the diagnosis of&#160;central fatigue. Changes of the central neurotransmitters also verified the result. In conclusion, the model successfully simulated central fatigue, and future study with the model may help reveal the pathological mechanism of the disease.

Here, we present a protocol to introduce a rat model of central fatigue using the modified multiple platform method (MMPM).

Modelo de una rata de la fatiga Central mediante la método de plataforma múltiple

In this article, we introduced a rat model of central fatigue using the modified multiple platform method (MMPM). The Multiple Platform box was designed ...

A Preclinical Model of Exertional Heat Stroke in Mice

Heat stroke is the most severe manifestation of heat-related illnesses. Classic heat stroke (CHS), also known as passive heat stroke, occurs at rest, whereas exertional heat stroke (EHS) occurs during physical activity. EHS differs from CHS in etiology, clinical presentation, and sequelae of multi-organ dysfunction. Until recently, only models of CHS have been well established. This protocol aims to provide guidelines for a refined preclinical mouse model of EHS that is free from major limiting factors such as the use of anesthesia, restraint, rectal probes, or electric shock. Male and female C57Bl/6 mice, instrumented with core temperature (Tc) telemetric probes were utilized in this model. For familiarization with the running mode, mice undergo 3 weeks of training using both voluntary and forced running wheels. Thereafter, mice run on a forced wheel inside a climatic chamber set at 37.5 &#176;C and 40%-50% relative humidity (RH) until displaying symptom limitation (e.g., loss of consciousness) at Tc of 42.1-42.5 &#176;C, although suitable results can be obtained at chamber temperatures between 34.5-39.5 &#176;C and humidity between 30%-90%. Depending on the desired severity, mice are removed from the chamber immediately for recovery in ambient temperature or remain in the heated chamber for a longer duration, inducing a more severe exposure and a higher incidence of mortality. Results are compared with sham-matched exercise controls (EXC) and/or na&#239;ve controls (NC). The model mirrors many of the pathophysiological outcomes observed in human EHS, including loss of consciousness, severe hyperthermia, multi-organ damage as well as inflammatory cytokine release, and acute phase responses of the immune system. This model is ideal for hypothesis-driven research to test preventative and therapeutic strategies that may delay the onset of EHS or reduce the multi-organ damage that characterizes this manifestation.

The protocol describes the development of a standardized, repeatable, preclinical model of exertional heat stroke (EHS) in mice free from adverse external stimuli such as electric shock. The model provides a platform for mechanistic, preventative, and therapeutic studies.

Un modelo preclínico de golpe de calor por esfuerzo en ratones

Heat stroke is the most severe manifestation of heat-related illnesses. Classic heat stroke (CHS), also known as passive heat stroke, occurs at rest, ...

Método de tubo de choque para modelar explosiones de bajo nivel altamente repetitivas en ratones

Modeling Highly Repetitive Low-level Blast Exposure in Mice

Exposure to explosive blasts is a significant risk factor for brain trauma among exposed persons. Although the effects of large blasts on the brain are well understood, the effects of smaller blasts such as those that occur during military training are less understood. This small, low-level blast exposure also varies highly according to military occupation and training tempo, with some units experiencing few exposures over the course of several years whereas others experience hundreds within a few weeks. Animal models are an important tool in identifying both the injury mechanisms and long-term clinical health risks following low-level blast exposure. Models capable of recapitulating this wide range of exposures are necessary to inform acute and chronic injury outcomes across these disparate risk profiles.
Although outcomes following a few low-level blast exposures are easily modeled for mechanistic study, chronic exposures that occur over a career may be better modeled by blast injury paradigms with repeated exposures that occur frequently over weeks and months. Shown here are methods for modeling highly repetitive low-level blast exposure in mice. The procedures are based on established and widely used pneumatic shocktube models of open-field blast exposure that can be scaled to adjust the overpressure parameters and the number or interval of the exposures. These methods can then be used to either enable mechanistic investigations or recapitulate the routine blast exposures of clinical groups under study.

Autor destacado: Descifrando los efectos a largo plazo de las exposiciones a explosiones de bajo nivel en ratones

Presented here are methods for producing repeated low-intensity blast exposures using mice.

Modelado de la exposición a explosiones de bajo nivel altamente repetitivas en ratones

Exposure to explosive blasts is a significant risk factor for brain trauma among exposed persons. Although the effects of large blasts on the brain are ...

Un modelo de ratón de la fatiga inducida por irradiación periférica

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Establishment of a Robust and Reproducible Model of Radiation-Induced Skin and Muscle Fibrosis

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