We thank Gilles Mithieux and Aude Barataud (INSERM U1213, Lyon, France) and Marie Liabeuf and Stephanie Lemarchand-Minde (Animal facility, l'Institut du Thorax, Nantes, France) for their help with the animal care protocol. This work was supported by grants from La r&#233;gion des Pays de la Loire, the Fondation d'Avenir, and the Casden Bank. We would like to thank Catherine Postic, Fadila Benhamed and Michelle Ca&#252;zac from l'institut Cochin for their hospitality and their help during the filming process.

Y alojamiento de animales: Obtener C57BL / 6 ratones machos de 8 semanas de edad. A las 10 semanas de edad, dar a los ratones C57BL / 6 libre acceso al agua y una dieta alta en grasas (DIO dieta: 35% kcal de la grasa, el 25,8% de caseína, 1,30% AIN mineral, 1,30% de vitaminas, 1,70% de fosfato dicálcico, 0,7 carbonato% de calcio, 2,10% citrato de potasio, 0,026% bitartrato de colina, un 8,9% de sacarosa, 0,384% cistina, 6,5% de celulosa, el 31,7% de manteca de cerdo, aceite de soja 3,3%, y 16.29% de maltodextrina) de 8 (SG) o 14 (BGYR) semanas antes de la cirugía bariátrica. Para promover el aumento de peso, dar a los ratones dedicada a la cirugía DGYR (farsa y DGYR) agua que contiene 20% de fructosa (w / v), además de la dieta alta en grasas. Ética Declaración: Todos los pasos del protocolo se describen a continuación sigue las directrices del Comité de Ética de Experimentación Animal de Pays de la Loire bajo el número de homologación de 01.953,01. 1. Preparación preoperatoria general <ol><li> Ayunar a los ratones durante 6 h antes de la cirugía. Gel diy se le dio comida de 3 días antes de la cirugía y exclusivamente un día antes de la cirugía. Inducir la anestesia en una cámara con 5% de isoflurano (0,4 L / min) y O 2 (dioxígeno; 0,4 L / min). Realizar una prueba de dedo del pie-pellizco para confirmar que la anestesia es eficaz. Administrar analgésicos (0,1 mg / kg de buprenorfina), antibióticos (10 mg / kg marbofloxacine), y pro-cinética (1 mg / kg de metoclopramida) a través de inyecciones subcutáneas. </li><li> Asignar un área de trabajo específica para los procedimientos quirúrgicos (distantes de los fans de tráfico de laboratorio y de ventilación). Limpiar la zona quirúrgica con desinfectantes de superficies duras reservados. Coloque una almohadilla térmica en el área de espacio de trabajo para mantener homeotermia ratón durante la cirugía. Aplicar una almohadilla inferior absorbente y limpia sobre el cojín eléctrico. </li><li> Afeitarse el abdomen desde el esternón hasta la pelvis usando una máquina de afeitar eléctrica. Envolver el abdomen del ratón en una envoltura de plástico esterilizada (Figuras 1A y 2A) y colocar el puntero del ratón en la posición supina sobre el calentamiento panuncio. </li><li> Aplique un ungüento veterinario en los ojos para evitar la sequedad, mientras que bajo anestesia. Coloque el ratón en el cono de la nariz y mantener la anestesia con isoflurano al 2% (0,4 L / min) y O 2 (0,4 L / min). </li><li> Antes de comenzar la cirugía, se puso una bata estéril examen, una gorra batas desechables, mascarillas y guantes estériles. Abrir un paquete de autoclave que contiene un conjunto instrumento quirúrgico esterilizado, que se requiere para todos los pasos de la cirugía. Cortar una ventana en la envoltura de plástico esterilizada y desinfectar la piel con 2 soluciones sucesivas de povidona-yodo. </li></ol> 2. Manga Gástrica: Procedimiento quirúrgico <ol><li> Laparotomía media (Figura 1B) <ol><li> Bajo un microscopio binocular y con un escalpelo, realizar una incisión en la línea media desde el esternón hasta la mitad del abdomen para abrir la cavidad abdominal (no dañar los músculos abdominales). Proteger la piel con una compresa estéril empapada con solución salina C 37 °. </li></ol></li><li> La exposición del estómago (Figura 1C) <ol><li> movilizar suavemente el estómago utilizando hisopos de algodón humedecido. Liberar el estómago de sus laterales estrechos adjuntos tejido conectivo utilizando hisopos de algodón humedecidos o micro tijeras. </li><li> Suavemente, externalizar el estómago completamente y colocar un colágeno hemostático comprimir detrás del estómago. Suturar el píloro y vasos esófago a lo largo de la curvatura mayor del estómago con 8,0 suturas no absorbibles para evitar futuras hemorragia (Figura 1D y E). Nota: El uso hemostático puede inducir daño tisular, asegúrese de utilizar solamente en los tejidos que tienen como objetivo ser resecados. </li></ol></li><li> Resección de la región cardiaca del estómago (fondo) <ol><li> Realizar una gastrotomía en la línea anatómica presente entre la región pilórica (corpus) y la región cardiaca de estómago (fondo) (Figura 1F). </li><li> Retire el alimento residual con 2 hisopos de algodón humedecidos hacia cada unolado del estómago. El uso de micro tijeras, corte el estómago a lo largo de la frontera entre el fundus y el corpus del estómago (Figura 1G). </li><li> Presione un colágeno hemostático comprimir durante 2 minutos en caso de hemorragia grave. Cerrar la abertura del estómago con 8,0 suturas no absorbibles de la unión gastro-esofágico hasta el final de la incisión (Figura 1H). </li></ol></li><li> La resección de la región pilórica del estómago (corpus) <ol><li> Completar la resección del estómago a lo largo de la curvatura mayor con el fin de eliminar aproximadamente 80% del estómago (Figura 1I). Estandarizar el ancho de la SG a 3 mm. Presione un colágeno hemostático comprimir durante 2 minutos en caso de hemorragia grave. </li><li> Cerrar la abertura de la región pilórica del estómago con 8,0 sutura no absorbible (Figura 1J). rodar suavemente 2 hisopos de algodón humedecidos hacia cada lado del estómago para asegurar que la sutura es a prueba de fugas. </li><li>Completar con 8,0 puntos de sutura no absorbibles en caso de fuga. Retire las compresas de colágeno hemostáticos y devolver el estómago a la cavidad abdominal. Deja un colágeno hemostático comprimir contra la sutura. </li></ol></li><li> cierre abdominal <ol><li> Cierre la capa muscular de la pared abdominal con 5,0 suturas no absorbibles (Figura 1C). Reducir la anestesia mediante la reducción de la concentración de isoflurano al 1%. Cierre la piel con 5,0 suturas no absorbibles (Figura 1 l). </li></ol></li></ol> 3. Roux-en-Y bypass gástrico: Procedimiento quirúrgico <ol><li> laparotomía media <ol><li> Bajo un microscopio binocular y con un escalpelo, realizar una incisión en la línea media desde el esternón hasta la mitad del abdomen para abrir la cavidad abdominal (no dañar los músculos abdominales). Proteger la piel con una compresa estéril empapada con 37 ° C una solución salina (Figura 2B). </li></ol> &lt;/ Li&gt; <li> Asa biliopancreática y asa alimentaria <ol><li> Exteriorizar el intestino (Figura 2C). Medida 8 cm del píloro y realizar dos ligaduras del intestino con 5,0 suturas no absorbibles (Figura 2D). </li><li> Cortar el intestino entre las dos ligaduras (Figura 2E). Coloque la extremidad proximal de los dos extremos en el cuadrante superior izquierdo del abdomen. Nota: Esto se utiliza como el asa alimentaria (Figura 2F). </li><li> Coloque la extremidad distal de los dos extremos que se enfrenta el asa alimentaria 6 cm por debajo de la extremidad proximal (Figura 2F). NOTA: Este será utilizado como el miembro biliar. </li></ol></li><li> Yeyuno-yeyunostomía <ol><li> Cortar tanto la extremidad proximal y el bucle de intestino utilizando micro tijeras y realizar dos incisiones antimesenteric de la misma longitud (Figura 2G y 2H). </li><li> Realizar lado a lado anastomosis con dos 8,0Las suturas no absorbibles. Realizar la anastomosis lado dorsal primero (Figura 2I), seguida de la anastomosis lado ventral (Figura 2J). </li><li> rodar suavemente 2 hisopos de algodón humedecidos hacia cada lado de la anastomosis para garantizar que la sutura es a prueba de fugas. Completar con 8,0 puntos de sutura no absorbibles en caso de fuga. </li></ol></li><li> Gastro-yeyunostomía <ol><li> movilizar suavemente el estómago utilizando hisopos de algodón humedecidos. Liberar el estómago de sus laterales estrechos adjuntos tejido conectivo utilizando hisopos de algodón humedecidos o micro tijeras. </li><li> Suavemente, exteriorizar el estómago completamente, la colocación de un re-absorbible colágeno hemostático comprimir detrás del estómago. Realizar una ligadura del píloro utilizando una sutura no absorbible 5.0 pasado a través del epiplón utilizando pinzas micro curvas (Figura 2K y 2L). </li><li> Cortar tanto en el lado ventral del estómago, 1,5 cm desde el píloro, y la extremidad distalutilizando micro tijeras, creando dos incisiones de la misma longitud (Figura 2 M). </li><li> Realizar la anastomosis de lado a lado con dos 8,0 suturas no absorbibles. Comience con la anastomosis lado dorsal (Figura 2 N), y luego realizar la anastomosis lado ventral (Figura 2O). rodar suavemente 2 hisopos de algodón humedecido hacia cada lado de la anastomosis para garantizar que la sutura es a prueba de fugas. Completar con 8,0 puntos de sutura no absorbibles en caso de fuga. </li></ol></li><li> cierre abdominal <ol><li> Cierre la capa muscular de la pared abdominal usando 5,0 suturas no absorbibles (Figura 2P). Reducir la anestesia mediante la reducción de la concentración de isoflurano al 1%. Cierre la piel utilizando 5.0 suturas no absorbibles (Figura 2Q) .Administer 25 ml / kg de solución salina caliente a través de la inyección subcutánea. </li></ol></li></ol> 4. general Cuidado posoperatorio <ol><li> Deja de isofluranoy continuar con un flujo de O2 de 0,8 l / min hasta que el ratón está totalmente despierto. No deje desatendido el ratón hasta que recupera el control motor, indicado por la partida de los animales a moverse alrededor de la jaula y ser capaz de ponerse de pie y caminar sin caerse. </li><li> Coloque el ratón (sólo un ratón por jaula) en una incubadora bajo condición de temperatura de 30 ° C durante 5 días. </li><li> Mantener hierro (0,5 mg / kg / día; inyección subcutánea) y vitamina (800 mg / 180 ml en agua) supplementations en los ratones DGYR hasta el final del protocolo. </li><li> Devolver el libre acceso a la dieta gel de alimentos (dieta con gel de alto contenido de grasa: 10% de manteca de cerdo, azúcar líquido al 10%, el 57% de agua) durante 5 días después de la cirugía. Reintroducir una dieta sólida de 3 días después de la cirugía. </li><li> Por vía subcutánea inyectar buprenorfina (0,1 mg / kg, dos veces al día, desde el día 0 al día 3 después de la cirugía), meloxicam (1 mg / kg, desde el día 0 al día 3 después de la cirugía), la metoclopramida (1 mg / kg, desde el día 0 a 5 días después de la cirugía), y marbofloxacine (10 mg / kg, desde el día 0 hasta el día 3 faet<html> cirugía r). </li></ol> 5. La evaluación postoperatoria metabólico Parámetros <ol><li> Medición de la ingesta de alimentos <ol><li> Permitir que los ratones se recuperen durante 1 semana después de la cirugía. Casa los ratones uno por jaula. Medir la cantidad de dieta sólida colocado en la jaula. El peso de los alimentos restantes 24 h más tarde. Repita este paso todo el tiempo necesario. </li></ol></li><li> Test oral de tolerancia a la glucosa <ol><li> Ayunar a los ratones durante 6 horas antes del bolo oral. Cosecha de una gota de sangre de la punta de la cola y aplicarlo sobre una tira de glucosa insertado en un glucómetro para determinar el valor de glucosa en sangre 0 tiempo. </li><li> Administrar un bolo oral de solución de D-glucosa 20% (2 g / kg). Medir los niveles de glucosa en sangre a los 15, 30, 60, y 120 minutos después de la alimentación forzada. </li></ol></li><li> El análisis del nivel de hemoglobina <ol><li> El uso de un sistema de hematología, medir 20 l de sangre fresca para probar el nivel de hemoglobina. </li></ol></li></ol>&quot;&gt; 6. La eutanasia <ol><li> Anestesiar a los ratones por vía intraperitoneal inyección de una solución de xilazina / quetamina (10/80 mg / kg). Realizar una dislocación cervical. Verificar la eutanasia completa, garantizando que el corazón no late y se pierde el reflejo de parpadeo globo ocular. </li></ol>

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The authors declare no conflicts of interest.

Para superar la creciente epidemia de obesidad, los primeros procedimientos de cirugía bariátrica surgieron en la década de 1960 en los Estados Unidos. Desde entonces, el número de procedimientos realizados en todo el mundo cada año aún aumentar, y hoy en día, estas técnicas representan la mejor opción terapéutica para el tratamiento de la obesidad mórbida 6. Entre los procedimientos desarrollados, SG y DGYR son los dos métodos más populares utilizados en la práctica clínica <sup c...

La pandemia mundial de la obesidad y la diabetes es devastador en gravedad. Más de dos mil millones de adultos en todo el mundo (30% de la población) tienen sobrepeso (IMC&gt; 25 kg / m 2) u obesidad (IMC&gt; 30 kg / m 2) 1. Esto puede venir junto con las complicaciones metabólicas, como la diabetes tipo 2, hipertensión y dislipidemia, lo que lleva a un aumento de la morbilidad y la mortalidad. La obesidad aumenta la mortalidad global y la prevalencia de cáncer 2. Debido a la falta de tratamientos no invasivos eficaces, la cirugía bariátrica representa la única opción que puede conducir a largo plazo, la pérdida de peso sostenida 3, 4. Un número de diferentes métodos quirúrgicos se han desarrollado, pero la gastrectomía en manga (SG) y el bypass gástrico Roux-en-Y (BGYR) son los dos procedimientos más utilizados en la práctica clínica. Durante el procedimiento de SG, 80% del volumen inicial del estómago esremoto; Por lo tanto, esta técnica es una de las cirugías restrictivas que mejora la sensación de saciedad. El BGYR es una de las técnicas restrictivas-malabsorción. Durante DGYR, se crea una pequeña bolsa gástrica (1-2% del volumen gástrico total) y el intestino se reordena en una forma de Y, lo que retrasa la digestión y la absorción de nutrientes. Estas dos técnicas conducen a una reducción significativa del peso corporal y la mejora general de las comorbilidades asociadas con frecuencia (por ejemplo, la hipertensión, la diabetes tipo 2 y dislipidemia) 3, con una mayor eficiencia se ve en DGYR. Sin embargo, los mecanismos moleculares detrás de los efectos beneficiosos pleiotrópicos de las cirugías bariátricas a menudo no están completamente aclarada. Debido a las numerosas cepas genéticamente modificadas disponibles, un modelo de ratón es el modelo animal más conveniente para explorar estos mecanismos moleculares. Sin embargo, los procedimientos bariátricos son difíciles de adaptar directamente a los pequeños modelos animales y requirE High destreza quirúrgica. Mientras SG se puede realizar fácilmente en roedores con una muy buena tasa de supervivencia, DGYR es letal en ratones debido a las obstrucciones intestinales graves 5. Se han propuesto diferentes técnicas de BGYR modificados para contrarrestar este problema, en particular el oesojejunostomy 5. A continuación, presentamos otra alternativa: la gastroyeyunostomía sin extirpación del estómago. Este DGYR modificado reproduce la mayor parte de los efectos beneficiosos observados en los seres humanos (es decir, una reducción de peso corporal significativa y una mejora de la glucosa y de lípidos homeostasis). Este manuscrito tiene como objetivo resumir y discutir los detalles técnicos y experimentales de SG y DGYR en ratones y facilitar estos procedimientos con la ayuda de vídeos. Se hará un destacado específica con respecto a la optimización de los protocolos sanitarios preoperatorios y postoperatorios que permiten la reducción de las vitaminas y de hierro.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Drugs</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>High Fat diet</td>
			<td>DIO diet</td>
			<td>Safe</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Forane</td>
			<td>Baxter</td>
			<td></td>
		</tr>
		<tr>
			<td>Buprenorphin</td>
			<td>Buprecare</td>
			<td>Animalcare</td>
			<td></td>
		</tr>
		<tr>
			<td>Marbofloxacine</td>
			<td>Marbocyl</td>
			<td>Vetoquinol</td>
			<td></td>
		</tr>
		<tr>
			<td>Ammonium iron citrate, vitamins PP-B12</td>
			<td>Fercobsang</td>
			<td>Vetoquinol</td>
			<td></td>
		</tr>
		<tr>
			<td>Vitamins A-D3-E-K-B</td>
			<td>Vita Rongeur</td>
			<td>Virbac</td>
			<td></td>
		</tr>
		<tr>
			<td>NaCl 0,9%</td>
			<td>NaCl 0,9%</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Povidone solution</td>
			<td>Betadine Scrub</td>
			<td>Betadine</td>
			<td></td>
		</tr>
		<tr>
			<td>Povidone solution</td>
			<td>Betadine Solution</td>
			<td>Betadine</td>
			<td></td>
		</tr>
		<tr>
			<td>Carboptol 980 NF</td>
			<td>Ocrygel</td>
			<td>TVM</td>
			<td></td>
		</tr>
		<tr>
			<td>Name</td>
			<td>References</td>
			<td>Company</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Sutures</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Prolene&#174;</td>
			<td>8.0, 6,5 mm</td>
			<td>Ethicon</td>
			<td></td>
		</tr>
		<tr>
			<td>Prolene&#174;</td>
			<td>5.0, 13 mm</td>
			<td>Ethicon</td>
			<td></td>
		</tr>
		<tr>
			<td>Name</td>
			<td>References</td>
			<td>Company</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Surgical equipments</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Scissors</td>
			<td></td>
			<td>FST</td>
			<td></td>
		</tr>
		<tr>
			<td>Needle holder</td>
			<td>Olsen-Hegar</td>
			<td>FST</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro scissors</td>
			<td>Vannas</td>
			<td>FST</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro forceps&#160;</td>
			<td>Graefe</td>
			<td>FST</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro forceps curved</td>
			<td>Graefe</td>
			<td>FST</td>
			<td></td>
		</tr>
		<tr>
			<td>Curved micro needle holder</td>
			<td>Castroviejo</td>
			<td>FST</td>
			<td></td>
		</tr>
		<tr>
			<td>Hemostatic collagen compress</td>
			<td>Pangen</td>
			<td>Urgo</td>
			<td></td>
		</tr>
		<tr>
			<td>Absorbent underpads</td>
			<td></td>
			<td>VWR</td>
			<td></td>
		</tr>
		<tr>
			<td>Name</td>
			<td>References</td>
			<td>Company</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Specific equipments</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Hematology system&#160;</td>
			<td>Hemavet 950FS</td>
			<td>Hemavet</td>
			<td></td>
		</tr>
		<tr>
			<td>Glucose strips and glucometer</td>
			<td>One touch Verio</td>
			<td>Life scan</td>
			<td></td>
		</tr>
		<tr>
			<td>Stereo microscope</td>
			<td>MZ6</td>
			<td>Leica&#160;</td>
			<td></td>
		</tr>
	</tbody>
</table>

techniques of sleeve gastrectomy and modified roux-en-y gastric bypass in mice

La obesidad es un importante problema de salud pública, con una prevalencia de 4 a 28% para los hombres y 6.2 a 36.5% para las mujeres en Europa (de 2003 a 2008). La obesidad mórbida se asocia con frecuencia a complicaciones metabólicas, como la diabetes tipo 2, hipertensión y dislipidemia, la reducción de la esperanza de vida y la calidad. En la ausencia de tratamientos no invasivos eficaces, la cirugía bariátrica es una valiosa opción terapéutica para los pacientes con obesidad mórbida (índice de masa corporal (IMC)&gt; 40 kg / m 2), que conduce a largo plazo, la pérdida de peso y la mejora de las complicaciones metabólicas sostenida . Sin embargo, los mecanismos celulares y moleculares subyacentes que sostienen los efectos beneficiosos de la cirugía bariátrica aún no se entienden completamente. Debido a las numerosas cepas genéticamente modificadas disponibles, el modelo de ratón es el modelo animal más conveniente para explorar los mecanismos moleculares detrás de los efectos beneficiosos pleiotrópicos de la cirugía bariátrica. A continuación, detallamos la Healthc optimizadoson métodos y protocolos quirúrgicos en los ratones de los dos cirugías bariátricas más ampliamente utilizados: la manga gástrica y el bypass gástrico Roux-en-Y modificada. Descifrar los mecanismos moleculares que subyacen a los efectos terapéuticos de la cirugía bariátrica ofrece la promesa de la identificación de nuevas dianas terapéuticas.

Condiciones generales El tiempo operatorio para el procedimiento de SG fue de 49,3 ± 1,5 min. Hemos eliminado 62,8 ± 5,0 mg de estómago, lo que representa alrededor del 80% por ciento del estómago. Sin ratones murieron durante la cirugía o durante los siguientes siete días. Un ratón (7,1%) murieron en el 11 º día postoperatorio debido a una obstrucción gástrica causada por un bezoar. ...

Watch this Scientific Journal Video about Techniques of Sleeve Gastrectomy and Modified Roux-en-Y Gastric Bypass in Mice at JoVE.com

Techniques of Sleeve Gastrectomy and Modified Roux-en-Y Gastric Bypass in Mice

Bariatric surgery is the most efficient way to reduce body weight and the deadly metabolic complications (diabetes, obesity, and dyslipidemia) frequently associated with morbid obesity. Mouse models of bariatric surgery represent a unique asset for deciphering molecular mechanisms behind the beneficial effects of these surgeries on diabetes, hypertension, and dyslipidemia.

Técnicas de Manga Gástrica y modificación de Roux-en-Y bypass gástrico en ratones

Obesity is a major public health issue, with a prevalence of 4 to 28% for men and 6.2 to 36.5% for women in Europe (from 2003 to 2008). Morbid obesity is ...

techniques-sleeve-gastrectomy-modified-roux-en-y-gastric-bypass

Research

JoVE Journal

Medicine

11.9K vistas.  INSERM, CNRS, UNIV Nantes. Bariatric surgery is the most efficient way to reduce body weight and the deadly metabolic complications (diabetes, obesity, and dyslipidemia) frequently associated with morbid obesity. Mouse models of bariatric surgery represent a unique asset for deciphering molecular mechanisms behind the beneficial effects of these surgeries on diabetes, hypertension, and dyslipidemia.

Video: Techniques of Sleeve Gastrectomy and Modified Roux-en-Y Gastric Bypass in Mice

Modified Technique for Coronary Artery Ligation in Mice

Myocardial infarction (MI) is one of the most important causes of mortality in humans1-3. In order to improve morbidity and mortality in patients with MI we need better knowledge about pathophysiology of myocardial ischemia. This knowledge may be valuable to define new therapeutic targets for innovative cardiovascular therapies4. Experimental MI model in mice is an increasingly popular small-animal model in preclinical research in which MI is induced by means of permanent or temporary ligation of left coronary artery (LCA)5. In this video, we describe the step-by-step method of how to induce experimental MI in mice.
The animal is first anesthetized with 2% isoflurane. The unconscious mouse is then intubated and connected to a ventilator for artificial ventilation. The left chest is shaved and 1.5 cm incision along mid-axillary line is made in the skin. The left pectoralis major muscle is bluntly dissociated until the ribs are exposed. The muscle layers are pulled aside and fixed with an eyelid-retractor. After these preparations, left thoracotomy is performed between the third and fourth ribs in order to visualize the anterior surface of the heart and left lung. The proximal segment of LCA artery is then ligated with a 7-0 ethilon suture which typically induces an infarct size ~40% of left ventricle. At the end, the chest is closed and the animals receive postoperative analgesia (Temgesic, 0.3 mg/50 ml, ip). The animals are kept in a warm cage until spontaneous recovery.

The surgical procedure used to induce experimental myocardial infarction in mice begins with left thoracotomy between the third and the fourth ribs in order to visualize the anterior surface of the heart and left lung. The left coronary artery is ligated, the chest is closed and the mouse is allowed to recover spontaneously.

Técnica modificada para la ligadura de la arteria coronaria en ratones

Myocardial infarction (MI) is one of the most important causes of mortality in humans1-3. In order to improve morbidity and mortality in patients with MI ...

Investigación

Medicina

Roux-en-Y Gastric Bypass Operation in Rats

Currently, the most effective therapy for the treatment of morbid obesity to induce significant and maintained body weight loss with a proven mortality benefit is bariatric surgery1,2. Consequently, there has been a steady rise in the number of bariatric operations done worldwide in recent years with the Roux-en-Y gastric bypass (gastric bypass) being the most commonly performed operation3. Against this background, it is important to understand the physiological mechanisms by which gastric bypass induces and maintains body weight loss. These mechanisms are yet not fully understood, but may include reduced hunger and increased satiation4,5, increased energy expenditure6,7, altered preference for food high in fat and sugar8,9, altered salt and water handling of the kidney10 as well as alterations in gut microbiota11. Such changes seen after gastric bypass may at least partly stem from how the surgery alters the hormonal milieu because gastric bypass increases the postprandial release of peptide-YY (PYY) and glucagon-like-peptide-1 (GLP-1), hormones that are released by the gut in the presence of nutrients and that reduce eating12.
During the last two decades numerous studies using rats have been carried out to further investigate physiological changes after gastric bypass. The gastric bypass rat model has proven to be a valuable experimental tool not least as it closely mimics the time profile and magnitude of human weight loss, but also allows researchers to control and manipulate critical anatomic and physiologic factors including the use of appropriate controls. Consequently, there is a wide array of rat gastric bypass models available in the literature reviewed elsewhere in more detail 13-15. The description of the exact surgical technique of these models varies widely and differs e.g. in terms of pouch size, limb lengths, and the preservation of the vagal nerve. If reported, mortality rates seem to range from 0 to 35%15. Furthermore, surgery has been carried out almost exclusively in male rats of different strains and ages. Pre- and postoperative diets also varied significantly.
Technical and experimental variations in published gastric bypass rat models complicate the comparison and identification of potential physiological mechanisms involved in gastric bypass. There is no clear evidence that any of these models is superior, but there is an emerging need for standardization of the procedure to achieve consistent and comparable data. This article therefore aims to summarize and discuss technical and experimental details of our previously validated and published gastric bypass rat model.

Numerous studies using gastric bypass rat models have been recently conducted to uncover the underlying physiological mechanisms of Roux-en-Y gastric bypass operations. This article aims to demonstrate and discuss the technical and experimental details of our published gastric bypass rat model to understand advantages and limitations of this experimental tool.

Roux-en-Y Gastric operación de bypass en las ratas

Currently, the most effective therapy for the treatment of morbid obesity to induce significant and maintained body weight loss with a proven mortality ...

Assessment of Gastric Emptying in Non-obese Diabetic Mice Using a [13C]-octanoic Acid Breath Test

Gastric emptying studies in mice have been limited by the inability to follow gastric emptying changes in the same animal since the most commonly used techniques require killing of the animals and postmortem recovery of the meal1,2. This approach prevents longitudinal studies to determine changes in gastric emptying with age and progression of disease. The commonly used [13C]-octanoic acid breath test for humans3 has been modified for use in mice4-6 and rats7 and we previously showed that this test is reliable and responsive to changes in gastric emptying in response to drugs and during diabetic disease progression8. In this video presentation the principle and practical implementation of this modified test is explained. As in the previous study, NOD LtJ mice are used, a model of type 1 diabetes9. A proportion of these mice develop the symptoms of gastroparesis, a complication of diabetes characterized by delayed gastric emptying without mechanical obstruction of the stomach10.
This paper demonstrates how to train the mice for testing, how to prepare the test meal and obtain 4 hr gastric emptying data and how to analyze the obtained data. The carbon isotope analyzer used in the present study is suitable for the automatic sampling of the air samples from up to 12 mice at the same time. This technique allows the longitudinal follow-up of gastric emptying from larger groups of mice with diabetes or other long-standing diseases.

Assessment of Gastric Emptying in Non-obese Diabetic Mice Using a [13C]-octanoic Acid Breath Test

Determination of gastric emptying with a non-invasive [13C]-octanoic acid breath test for tracking gastroparesis in female NOD LtJ mice.

Evaluación del vaciamiento gástrico en ratones diabéticos no obesos mediante un [<sup&gt; 13</sup&gt; C]-octanoico prueba del aliento ácido

Gastric emptying studies in mice have been limited by the inability to follow gastric emptying changes in the same animal since the most commonly used ...

Using the Sleeve Technique in a Mouse Model of Aortic Transplantation - An Instructional Video

Orthotopic aortic transplantation using the sleeve technique reduces injury to the aorta with failure rate of only 10-20%. The time to anastomose the aorta in mice using the sleeve method was short and easy averaging 20 min, permitting studies of iso/allo grafts. The following article describes the aortic transplantation procedure used in our laboratory. The mice were anesthetized with a mixture of 1.5% volume isoflurane and 100% oxygen through a face mask. At this point, the segment of the aorta between the renal arteries and its bifurcation was separated from the vena cava, freely prepared and clampedat the proximal and distal segments with a single silk suture. Prior to the removal of the aorta, a saline solution containing heparin was injected into the inferior vena cava. Then the aorta was cut between the clamps and a saline heparin solution was used to flush the lumen. The sleeve technique with monofilament sutures was used in order to transplant the abdominal aorta in the orthotopic position.

We present an orthotopic aortic transplantation model using the sleeve technique in mice. It is a very rapid anastomosis method, which can be employed in studies of vascular disease.

Utilizando la técnica de la manga en un modelo murino de trasplante aórtico - un Video instructivo

Orthotopic aortic transplantation using the sleeve technique reduces injury to the aorta with failure rate of only 10-20%. The time to anastomose the ...

Cardiopulmonary Bypass in a Mouse Model: A Novel Approach

As prolonged cardiopulmonary bypass becomes more essential during cardiac interventions, an increasing clinical demand arises for procedure optimization and for minimizing organ damage resulting from prolonged extracorporal circulation. The goal of this paper was to demonstrate a fully functional and clinically relevant model of cardiopulmonary bypass in a mouse. We report on the device design, perfusion circuit optimization, and microsurgical techniques. This model is an acute model, which is not compatible with survival due to the need for multiple blood drawings. Because of the range of tools available for mice (e.g., markers, knockouts, etc.), this model will facilitate investigation into the molecular mechanisms of organ damage and the effect of cardiopulmonary bypass in relation to other comorbidities.

This paper describes how to perform cardiopulmonary bypass in mice. This novel model will facilitate the investigation of the molecular mechanisms involved in organ damage.

Puente cardiopulmonar en un modelo de ratón: un nuevo enfoque

As prolonged cardiopulmonary bypass becomes more essential during cardiac interventions, an increasing clinical demand arises for procedure optimization ...

One-anastomosis Gastric Bypass (OAGB) in Rats

The goal of this protocol is to set up a preclinical model of bariatric surgery and, more specifically, OAGB in obese rats. Based on this preclinical model, longitudinal studies can be carried out to provide an improved understanding of the mechanisms underlying the outcomes seen after bariatric surgery in humans. For this purpose, rats are operated on through a laparotomy under general anesthesia with isoflurane. First, the surgeon creates a long and tubular gastric pouch: after greater curve and hiatal dissection, the nonglandular stomach is stapled and removed. Then, the remaining stomach is also stapled in order to create a gastric tube and exclude the antrum of the stomach. After that, the surgeon performs a single end-to-side gastrojejunostomy 35 cm from the duodenojejunal angle. This limb length has been chosen in order to reproduce the same ratio between the biliopancreatic limb (BPL) and common limb (CL) length as in human bariatric surgery. The operation ends by aponeurotic and cutaneous closure. The early postoperative management consists of subcutaneous hydration, an intramuscular prophylactic antibiotic injection, a parietal injection of xylocaine, the administration of painkillers, and a progressive reintroduction of diet.

One-anastomosis Gastric Bypass OAGB in Rats

This protocol is for the performance of one-anastomosis gastric bypass (OAGB) on rat. The operator performs a long and tubular-stapled gastric pouch followed by hand-sewn anastomosis. For this model, the operator reproduces the same ratio between biliopancreatic and common limb in humans; therefore, the biliopancreatic limb measures 35 cm.

Bypass gástrico de una anastomosis (OAGB) en ratas

The goal of this protocol is to set up a preclinical model of bariatric surgery and, more specifically, OAGB in obese rats. Based on this preclinical ...

Sleeve Gastrectomy in Mice using Surgical Clips

The number of people who are overweight and obese is continually increasing both in the adult and adolescent populations. This coincides with the increased universal phenomenon of type 2 diabetes (T2D) and other metabolic problems. Bariatric surgery, such as SG, is currently one of the most effective and commonly used long-term treatment for obesity and T2D, but the association between them is not completely explored yet. The mechanisms underlying the outcomes seen after bariatric surgery in humans can be investigated based on preclinical animal studies. The SG reduces body weight, glucose levels and many metabolic parameters, and is easy to perform with a low incidence of complications. The goal of this work is to provide a simple method and an uncomplicated preclinical model of bariatric surgery in animals for researchers.

The prevalence of diabetes and obesity is continuously increasing worldwide. The mechanisms between diabetes, obesity and their associated mortality and co-morbidities need to be further investigated. Here, we present a protocol for sleeve gastrectomy (SG) in animals as an uncomplicated preclinical model of bariatric surgery.

Gastrectomía de manga en ratones usando clips quirúrgicos

The number of people who are overweight and obese is continually increasing both in the adult and adolescent populations. This coincides with the ...

A Protocol for Roux-en-Y Gastric Bypass in Rats using Linear Staplers

Roux-en-Y gastric bypass (RYGB) is commonly performed for the treatment of severe obesity and type 2 diabetes. However, the mechanism of weight loss and metabolic changes are not well understood. Multiple factors are thought to play a role, including reduced caloric intake, decreased nutrient absorption, increased satiety, the release of satiety-promoting hormones, shifts in bile acid metabolism, and alterations in the gut microbiota.
The rat RYGB model presents an ideal framework to study these mechanisms. Prior work on mouse models have had high mortality rates, ranging from 17 to 52%, limiting their adoption. Rat models demonstrate more physiologic reserve to surgical stimulus and are technically easier to adopt as they allow for the use of surgical staplers. One challenge with surgical staplers, however, is that they often leave a large gastric pouch which is not representative of RYGB in humans. 
In this protocol, we present a RYGB protocol in rats that result in a small gastric pouch using surgical staplers. Utilizing two stapler fires which remove the forestomach of the rat, we obtain a smaller gastric pouch similar to that following a typical human RYGB. Surgical stapling also results in better hemostasis than sharp division. Additionally, the forestomach of the rat does not contain any glands and its removal should not alter the physiology of RYGB.
Weight loss and metabolic changes in the RYGB cohort were significant compared to the sham cohort, with significantly lower glucose tolerance at 14 weeks. Furthermore, this protocol has an excellent survival of 88.9% after RYGB. The skills described in this protocol can be acquired without previous microsurgical experience. Once mastered, this procedure will provide a reproducible tool for studying the mechanisms and effects of RYGB.

Roux-en-Y gastric bypass (RYGB) is performed to treat obesity and diabetes. However, the mechanisms underlying RYGB's efficacy are not fully understood, and studies are limited by technical difficulty leading to high mortality in animal models. This article provides instructions on how to perform RYGB in rats with high success rates.

Un Protocolo Para Roux-en-Y Bypass Gástrico En Ratas Usando Grapadoras Lineales

Roux-en-Y gastric bypass (RYGB) is commonly performed for the treatment of severe obesity and type 2 diabetes. However, the mechanism of weight loss and ...

Robotic Central Pancreatectomy with Roux-en-Y Pancreaticojejunostomy

Central pancreatectomy is a parenchyma-sparing alternative to distal pancreatectomy in patients with a benign or low-grade malignant tumor in the body of the pancreas. The aim of central pancreatectomy is to prevent postoperative life-long endocrine and exocrine insufficiency. The downside of central pancreatectomy is the high rate of postoperative pancreatic fistula, which is the main reason that many surgeons do not routinely use central pancreatectomy in eligible patients. Most studies report open or laparoscopic central pancreatectomy with a pancreatico-gastrostomy anastomosis in adults. This is the first description of a standardized approach to robotic central pancreatectomy with Roux-en-Y pancreaticojejunostomy reconstruction in an adolescent (16-year-old boy) with a pseudopapillary tumor in the body of the pancreas. The operation time was 248 min with 20 mL of blood loss. The postoperative course was uneventful except for the short-term medical treatment for a grade B pancreatic fistula. Robotic central pancreatectomy can be safely applied in selected patients in experienced centers.

Robotic central pancreatectomy may be used in selected patients in experienced centers. This protocol presents all steps and the feasibility of a robotic central pancreatectomy with Roux-en-Y pancreaticojejunostomy in a 16-year-old adolescent patient.

Pancreatectomía central robótica con pancreaticojeyunostomía Roux-en-Y

Central pancreatectomy is a parenchyma-sparing alternative to distal pancreatectomy in patients with a benign or low-grade malignant tumor in the body of ...

A Modified Simple Method for Induction of Myocardial Infarction in Mice

Myocardial infarction (MI) represents one of the leading causes of death. MI models are widely used for investigating the pathomechanisms of post-MI remodeling and evaluation of novel therapeutics. Different methods (e.g., isoproterenol treatment, cryoinjury, coronary artery ligation, etc.) have been used to induce MI. Compared with isoproterenol treatment and cryoinjury, coronary artery ligation may better reflect the ischemic response and chronic remodeling after MI. However, traditional methods for coronary ligation in mice are technically challenging. The current study describes a simple and efficient process for induction of MI in mice with readily available materials. The mouse chest skin was cut open under stable anesthesia. The heart was immediately externalized through the intercostal space after blunt separation of the pectoralis major and pectoralis minor. The left anterior descending branch (LAD) was ligated with a 6-0 suture 3 mm from its origin. Following LAD ligation, staining with 2,3,5-Triphenyltetrazolium chloride (TTC) indicated successful induction of MI and temporal changes of post-MI scar size. Meanwhile, survival analysis results showed overt mortality within 7 days after MI, mainly due to cardiac rupture. Moreover, post-MI echocardiographic assessment demonstrated successful induction of contractile dysfunction and ventricular remodeling. Once mastered, an MI model can be established in mice within 2-3 min with readily available materials.

Under adequate anesthesia, the mouse heart was externalized through the intercostal space, and myocardial infarction was successfully induced by ligating the left anterior descending artery (LAD) using materials readily available in most laboratories.

Un método simple modificado para la inducción de infarto de miocardio en ratones

Myocardial infarction (MI) represents one of the leading causes of death. MI models are widely used for investigating the pathomechanisms of post-MI ...