Name: mouse microsurgery infusion technique for targeted substance delivery into the cns via the internal carotid artery
Uploaded: 2017-01-31T14:00:00
Description: Watch this Scientific Journal Video about Mouse Microsurgery Infusion Technique for Targeted Substance Delivery into the CNS via the Internal Carotid Artery at JoVE.com

We would like to thank Dr. Lei Chen (Icahn School of Medicine at Mount Sinai, NY) who first established the use of this model in our laboratory, and to Dr. Gretchen Wolff (German Cancer Research Center, Heidelberg, Germany) for disseminating the technique in our laboratory. Supported in part by HL126559, DA039576, MH098891, MH63022, MH072567, DA027569, and NSC 2015/17/B/NZ7/02985.

Los procedimientos involucrados en el siguiente protocolo han sido aprobados por la Universidad de Miami Institucional Cuidado de Animales y el empleo Comisión (IACUC). Además, todos los procedimientos se llevan a cabo en las instalaciones aprobadas por la Asociación para la Evaluación y Acreditación de Laboratorio Animal Care International (AAALAC). 1. Preparación de los ratones para la cirugía <ol><li> Anestesiar al ratón con isoflurano mezclado con oxígeno, utilizando un sistem...

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La microcirugía de infusión descrito aquí se ha demostrado ser muy exitoso en la entrega de sustancias exógenas de diversas características biológicas en el SNC, la prevención de la difusión no deseada a través del cuerpo 1, 12. La interrupción de la barrera sangre-cerebro es una característica patológica de diversas enfermedades relacionadas con el SNC; por lo tanto, la evaluación de la relación de sustancias exógenas con la barrera sangre-cerebr...

En modelos in vivo del sistema nervioso central (SNC) requieren un suministro eficaz de sustancias exógenas, tales como medicamentos, agentes patógenos, o exosomas, en el cerebro. Por lo tanto, un método de entrega ideal debe causar un trauma mínimo para el animal, preservar la integridad de la red neuronal, y alcanzar altas concentraciones de sustancia en el cerebro 1. Se han descrito varios métodos quirúrgicos de entrega sustancia local, incluyendo intra-vaina, intracerebral, y las inyecciones intraventriculares o implantes 2, 3, 4, 5. Estos enfoques, sin embargo, se consideran traumática en el sistema nervioso central, y permiten la administración de sólo volúmenes bajos de la sustancia de interés. Por otra parte, se ha sugerido que las sustancias exógenas pueden eliminarse rápidamente por el líquido cefalorraquídeo 6 </sup&gt;, y una gama baja penetración en el parénquima cerebral se ha observado 7 cuando se emplean las técnicas antes mencionadas. Métodos de administración sistémicas, tales como administración oral, pulmonar, subcutánea, intravenosa, y se utilizan más comúnmente en modelos animales, a pesar de que exhiben una baja eficacia en la entrega de las sustancias en el sistema nervioso central, debido a la absorción por otros órganos 8, 9. Por lo tanto, estas vías de administración requieren dosis elevadas de las sustancias administradas, aumentando el riesgo de efectos secundarios y toxicidad 10, 11. A continuación, describimos una técnica de microcirugía infusión de ratón, que permite suministrar sustancias directamente en el cerebro a través de la arteria carótida interna. Además de dirigirse a la entrega al sistema nervioso central, esta técnica no pasa por alto las barreras fisiológicas normales y por lo tanto es de gran importancia para biological procesos involucrados en los pasajes de la terapéutica o patógenos en el cerebro.

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			<td height="21" style="height:21px;width:367px;">Anesthesia instrument</td>
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			<td align="right" style="width:119px;">901806</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:367px;">Surgical scissors</td>
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			<td height="21" style="height:21px;width:367px;">Surgical forceps straight tip</td>
			<td style="width:149px;">Fine Science Tool</td>
			<td style="width:119px;">00108-11</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:367px;">Surgical forceps angled tip</td>
			<td style="width:149px;">Fine Science Tool</td>
			<td style="width:119px;">00109-11</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:367px;">Spring scissors</td>
			<td style="width:149px;">Fine Science Tool</td>
			<td style="width:119px;">15000-08</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:367px;">Nylon suture</td>
			<td style="width:149px;">Braintree Scientific</td>
			<td style="width:119px;">SUT-S 104</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Capillary tubing (Micro-Renathane 0.010&#8221; x 0.005&#8221; per ft.)&#160;</td>
			<td style="width:149px;">Braintree Scientific</td>
			<td style="width:119px;">MRE01050</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:367px;">Closing suture</td>
			<td style="width:149px;">VWR</td>
			<td style="width:119px;">95057-036</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:367px;">Isoflurane</td>
			<td style="width:149px;">Piramal</td>
			<td style="width:119px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:367px;">2,3,5-Triphenyltetrazolium chloride</td>
			<td style="width:149px;">FisherScientific</td>
			<td style="width:119px;">50-121-8005</td>
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mouse microsurgery infusion technique for targeted substance delivery into the cns via the internal carotid artery

Animal models of central nervous system (CNS) diseases and, consequently, blood-brain barrier disruption diseases, require the delivery of exogenous substances into the brain. These exogenous substances may induce injurious impact or constitute therapeutic strategy. The most common delivery methods of exogenous substances into the brain are based on systemic deliveries, such as subcutaneous or intravenous routes. Although commonly used, these approaches have several limitations, including low delivery efficacy into the brain. In contrast, surgical methods that locally deliver substances into the CNS are more specific and prevent the uptake of the exogenous substances by other organs. Several surgical methods for CNS delivery are available; however, they tend to be very traumatic. Here, we describe a mouse infusion microsurgery technique, which effectively delivers substances into the brain via the internal carotid artery, with minimal trauma and no interference with normal CNS functionality.

La técnica de microcirugía infusión de ratón descrito aquí es muy versátil y se ha utilizado para suministrar diferentes sustancias directamente en el cerebro, incluyendo la entrega de células tumorales en un modelo representativo de la formación de metástasis cerebral 1, 12. Esta técnica también es adecuado para evaluar los aspectos patológicos de diferent...

Watch this Scientific Journal Video about Mouse Microsurgery Infusion Technique for Targeted Substance Delivery into the CNS via the Internal Carotid Artery at JoVE.com

Mouse Microsurgery Infusion Technique for Targeted Substance Delivery into the CNS via the Internal Carotid Artery

The present protocol describes a mouse microsurgery infusion technique, which effectively delivers substances directly into the brain via the internal carotid artery.

Técnica ratón Microcirugía de infusión para Targeted Sustancia de entrega en el SNC<i&gt; vía</i&gt; La arteria carótida interna

Animal models of central nervous system (CNS) diseases and, consequently, blood-brain barrier disruption diseases, require the delivery of exogenous ...

Research

JoVE Journal

Neuroscience

Mouse Microsurgery Infusion Technique for Targeted Substance Delivery into the CNS via the Internal Carotid Artery

Ultrasound-Guided Microinjection into the Mouse Forebrain In Utero at E9.5

Ultrasound-Guided Microinjection into the Mouse Forebrain In Utero at E9.5

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In utero survival surgery in mice permits the molecular manipulation of gene expression during development.  However, because the uterine wall is opaque ...

Investigación

Neurociencias

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