Name: methods for experimental manipulations after optic nerve transection in the mammalian cns
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Description: Watch this Scientific Journal Video about Methods for Experimental Manipulations after Optic Nerve Transection in the Mammalian CNS at JoVE.com

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1. Técnica quirúrgica <ol><li> Los experimentos deben llevarse a cabo utilizando una técnica aséptica y siguiendo los protocolos de uso de los animales de su institución específica. Instrumentos y materiales (soluciones, las sustancias químicas, marcadores, agujas, etc) que entran en contacto con el tejido vivo debe ser estéril para prevenir la infección y los efectos negativos sobre el bienestar animal y los posibles impactos negativos en el estudio. </li></ol> 2. Anestesia <ol><li>...

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	<li>Bahr, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Live+or+let+die+-+retinal+ganglion+cell+death+and+survival+during+development+and+in+the+lesioned+adult+CNS.">Live or let die - retinal ganglion cell death and survival during development and in the lesioned adult CNS.</a> Trends Neurosci. 23, 483-4890 (2000).</li><li>Isenmann, S., Kretz, A., Cellerino, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+determinants+of+retinal+ganglion+cell+development,+survival,+and+regeneration.">Molecular determinants of retinal ganglion cell development, survival, and regeneration.</a> Prog Retin Eye Res. 22, 483-543 (2003).</li><li>Koeberle, P. D., Bahr, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Growth+and+guidance+cues+for+regenerating+axons:+where+have+they+gone.">Growth and guidance cues for regenerating axons: where have they gone.</a> J Neurobiol. 59, 162-180 (2004).</li><li>Weishaupt, J. H., Bahr, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Degeneration+of+axotomized+retinal+ganglion+cells+as+a+model+for+neuronal+apoptosis+in+the+central+nervous+system+-+molecular+death+and+survival+pathways.">Degeneration of axotomized retinal ganglion cells as a model for neuronal apoptosis in the central nervous system - molecular death and survival pathways.</a> Restor. Neurol. Neurosci. 19, 1-2 (2001).</li><li>Arai-Gaun, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heme+oxygenase-1+induced+in+muller+cells+plays+a+protective+role+in+retinal+ischemia-reperfusion+injury+in+rats.">Heme oxygenase-1 induced in muller cells plays a protective role in retinal ischemia-reperfusion injury in rats.</a> Invest Ophthalmol Vis Sci. 45, 4226-4232 (2004).</li><li>Bainbridge, J. W., Tan, M. H., Ali, R. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gene+therapy+progress+and+prospects:+the+eye.">Gene therapy progress and prospects: the eye.</a> Gene Ther. 13, 1191-1197 (2006).</li><li>Polo, A. D. i. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prolonged+delivery+of+brain-derived+neurotrophic+factor+by+adenovirus-infected+Muller+cells+temporarily+rescues+injured+retinal+ganglion+cells.">Prolonged delivery of brain-derived neurotrophic factor by adenovirus-infected Muller cells temporarily rescues injured retinal ganglion cells.</a> Proc Natl Acad Sci U S A. 95, 3978-3983 (1998).</li><li>Herard, A. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=siRNA+targeted+against+amyloid+precursor+protein+impairs+synaptic+activity+in+vivo.">siRNA targeted against amyloid precursor protein impairs synaptic activity in vivo.</a> Neurobiol Aging. 27, 1740-1750 (2006).</li><li>Koeberle, P. D., Bahr, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+upregulation+of+GLAST-1+is+an+indirect+antiapoptotic+mechanism+of+GDNF+and+neurturin+in+the+adult+CNS.">The upregulation of GLAST-1 is an indirect antiapoptotic mechanism of GDNF and neurturin in the adult CNS.</a> Cell Death Differ. 15, 471-483 (2008).</li><li>Koeberle, P. D., Gauldie, J., Ball, A. K. <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+adenoviral-mediated+gene+transfer+of+interleukin-10,+interleukin-4,+and+transforming+growth+factor-beta+on+the+survival+of+axotomized+retinal+ganglion+cells.">Effects of adenoviral-mediated gene transfer of interleukin-10, interleukin-4, and transforming growth factor-beta on the survival of axotomized retinal ganglion cells.</a> Neuroscience. 125, 903-920 (2004).</li><li>Naik, R., Mukhopadhyay, A., Ganguli, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gene+delivery+to+the+retina:+focus+on+non-viral+approaches.">Gene delivery to the retina: focus on non-viral approaches.</a> Drug Discov Today. 14, 306-315 (2009).</li><li>van Adel, B. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Delivery+of+ciliary+neurotrophic+factor+via+lentiviral-mediated+transfer+protects+axotomized+retinal+ganglion+cells+for+an+extended+period+of+time.">Delivery of ciliary neurotrophic factor via lentiviral-mediated transfer protects axotomized retinal ganglion cells for an extended period of time.</a> Hum Gene Ther. 14, 103-115 (2003).</li><li>Alexander, J. J., Hauswirth, W. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adeno-associated+viral+vectors+and+the+retina.">Adeno-associated viral vectors and the retina.</a> Adv Exp Med Biol. 613, 121-128 (2008).</li><li>Allocca, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=AAV-mediated+gene+transfer+for+retinal+diseases.">AAV-mediated gene transfer for retinal diseases.</a> Expert Opin Biol Ther. 6, 1279-1294 (2006).</li><li>Surace, E. M., Auricchio, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Versatility+of+AAV+vectors+for+retinal+gene+transfer.">Versatility of AAV vectors for retinal gene transfer.</a> Vision Res. 48, 353-359 (2008).</li><li>Garcia-Valenzuela, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Axon-mediated+gene+transfer+of+retinal+ganglion+cells+in+vivo.">Axon-mediated gene transfer of retinal ganglion cells in vivo.</a> J Neurobiol. 32, 111-122 (1997).</li><li>Koeberle, P. D., Wang, Y., Schlichter, L. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kv1.1+and+Kv1.3+channels+contribute+to+the+degeneration+of+retinal+ganglion+cells+after+optic+nerve+transection+in+vivo.">Kv1.1 and Kv1.3 channels contribute to the degeneration of retinal ganglion cells after optic nerve transection in vivo.</a> Cell Death Differ. 17, 134-144 (2010).</li><li>Kugler, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transduction+of+axotomized+retinal+ganglion+cells+by+adenoviral+vector+administration+at+the+optic+nerve+stump:+an+in+vivo+model+system+for+the+inhibition+of+neuronal+apoptotic+cell+death.">Transduction of axotomized retinal ganglion cells by adenoviral vector administration at the optic nerve stump: an in vivo model system for the inhibition of neuronal apoptotic cell death.</a> Gene Ther. 6, 1759-1767 (1999).</li><li>Lingor, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Down-regulation+of+apoptosis+mediators+by+RNAi+inhibits+axotomy-induced+retinal+ganglion+cell+death+in+vivo.">Down-regulation of apoptosis mediators by RNAi inhibits axotomy-induced retinal ganglion cell death in vivo.</a> Brain. 128, 550-558 (2005).</li><li>Leon, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lens+injury+stimulates+axon+regeneration+in+the+mature+rat+optic+nerve.">Lens injury stimulates axon regeneration in the mature rat optic nerve.</a> J Neurosci. 20, 4615-4626 (2000).</li><li>Mansour-Robaey, S. <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+ocular+injury+and+administration+of+brain-derived+neurotrophic+factor+on+survival+and+regrowth+of+axotomized+retinal+ganglion+cells.">Effects of ocular injury and administration of brain-derived neurotrophic factor on survival and regrowth of axotomized retinal ganglion cells.</a> Proc Natl Acad Sci U S A. 91, 1632-1636 (1994).</li><li>D'Onofrio, P. M., Magharious, M. M., Koeberle, P. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optic+Nerve+Transection:+A+Model+of+Adult.">Optic Nerve Transection: A Model of Adult.</a> J Vis Exp. , .</li></ol>

Transección del nervio óptico es un modelo altamente reproducible de la apoptosis de las neuronas del SNC adulto. Las manipulaciones experimentales demostraron en este manuscrito permite el estudio de los mecanismos de apoptosis RGC después de la lesión. Inyecciones intraoculares son útiles para la orientación global de la retina. Este procedimiento requiere una cierta práctica, ya que es fundamental para no dañar la lente con la punta de la pipeta de vidrio. Daños en el objetivo ha...

A correction to the article Methods for Experimental Manipulations after Optic Nerve Transection in the Mammalian CNS was made.

Erratum: Methods for Experimental Manipulations after Optic Nerve Transection in the Mammalian CNS

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methods for experimental manipulations after optic nerve transection in the mammalian cns

Las células ganglionares de la retina (CGR) son las neuronas del SNC que la producción de información visual desde la retina hasta el cerebro, a través del nervio óptico. El nervio óptico se puede acceder dentro de la órbita del ojo y completamente seccionado (axotomized), el corte de los axones de la población RGC todo. Transección del nervio óptico es un modelo reproducible de la muerte apoptótica de las células neuronales en el SNC adulto 1-4. Este modelo es particularmente atractivo debido a que la cámara vítrea del ojo actúa como una cápsula para administración de fármacos a la retina, lo que permite manipulaciones experimentales a través de inyecciones intraoculares. La difusión de los productos químicos a través del humor vítreo se asegura de que actúan sobre la población RGC todo. Vectores virales, los plásmidos o corto RNAs de interferencia (siRNA) también puede ser entregado a la cámara vítrea para infectar o transfectar células de la retina 05.12. El tropismo de alta de virus adeno-asociado (AAV) vectores es beneficioso para la CGR de destino, con una tasa de infección cercana al 90% de las células cerca del sitio de la inyección 6, 7, 13-15. Por otra parte, la CGR puede ser selectiva transfectadas mediante la aplicación de siRNAs, plásmidos o vectores virales para el extremo del corte del nervio óptico o vectores 16-19 inyectar en su objetivo el colículo superior 10. Esto permite a los investigadores a estudiar los mecanismos de apoptosis en la población de neuronas lesionadas, sin factores de confusión en las neuronas o glía espectador otras circundantes. RGC apoptosis tiene una característica de tiempo-por supuesto por el que se retrasa la muerte celular postaxotomy 3-4 días, después de que las células degeneran rápidamente. Esto proporciona una ventana para la manipulación experimental dirigida contra los involucrados en la apoptosis. Manipulaciones que afectan directamente a la CGR del tronco del nervio óptico seccionado se llevan a cabo en el momento de axotomía, inmediatamente después de cortar el nervio. Por el contrario, cuando las sustancias son entregados a través de una vía intraocular, que puede ser inyectada antes de la cirugía o durante los primeros 3 días después de la cirugía, antes de la iniciación de la apoptosis en CGR axotomized. En el presente artículo, demostramos varios métodos para la manipulación experimental después de la transección del nervio óptico.

Watch this Scientific Journal Video about Methods for Experimental Manipulations after Optic Nerve Transection in the Mammalian CNS at JoVE.com

Methods for Experimental Manipulations after Optic Nerve Transection in the Mammalian CNS

Transección del nervio óptico es un modelo ampliamente utilizado de la lesión en el SNC adulto. Este modelo es ideal para llevar a cabo una serie de manipulaciones experimentales que se dirigen a la retina a nivel mundial o directamente dirigidos a la población heridos neuronal de las células ganglionares de la retina.

Los métodos para la manipulación experimental después de la transección del nervio óptico en el SNC de los mamíferos

Retinal ganglion cells (RGCs) are CNS neurons that output visual information from the retina to the brain, via the optic nerve. The optic nerve can be ...

Research

JoVE Journal

Neuroscience

Optic Nerve Transection: A Model of Adult Neuron Apoptosis in the Central Nervous System

La transección del nervio óptico: un modelo de apoptosis neuronal de adultos en el sistema nervioso central

Retinal ganglion cells (RGCs) are CNS neurons that output visual information from the retina to the brain, via the optic nerve. 
The optic nerve can be ...

Investigación

Neurociencias

An Optic Nerve Crush Injury Murine Model to Study Retinal Ganglion Cell Survival

Un aplastamiento del nervio óptico Lesiones modelo murino para estudiar la supervivencia de células ganglionares retinianas

Injury to the optic nerve can lead to axonal degeneration, followed by a gradual death of retinal ganglion cells (RGCs), which results in irreversible ...

Transplantation of Olfactory Ensheathing Cells to Evaluate Functional Recovery after Peripheral Nerve Injury

El trasplante de células ensheathing olfativo para evaluar la recuperación funcional después de la lesión del nervio periférico

Olfactory ensheathing cells (OECs) are neural crest cells which allow growth and regrowth of the primary olfactory neurons. Indeed, the primary olfactory ...

Motor Nerve Transection and Time-lapse Imaging of Glial Cell Behaviors in Live Zebrafish

Motor transección del nervio y Time-lapse de imágenes de Conductas de células gliales en el pez cebra en vivo

The  nervous system is often described as a hard-wired component of the body even  though it is a considerably fluid organ system that reacts to external ...

Fast Micro-iontophoresis of Glutamate and GABA: A Useful Tool to Investigate Synaptic Integration

Fast Micro-iontoforesis de glutamato y GABA: una herramienta útil para investigar la integración sináptica

One of the fundamental interests in neuroscience is to understand the integration of excitatory and inhibitory inputs along the very complex structure of ...

In vivo Imaging of Optic Nerve Fiber Integrity by Contrast-Enhanced MRI in Mice

In vivo Imaging of Optic Nerve Fiber Integrity by Contrast-Enhanced MRI in Mice

Imágenes in vivo de Óptica de Fibras Nerviosas Integridad de la RM con contraste en ratones

The rodent visual system encompasses retinal ganglion cells and their axons that form the optic nerve to enter thalamic and midbrain centers, and ...

Spinal Cord Transection in the Larval Zebrafish

La transección de la médula espinal en el pez cebra larval

Mammals fail in sensory and motor recovery following spinal cord injury due to lack of axonal regrowth below the level of injury as well as an inability ...

Minimally-invasive Technique for Injection into Rat Optic Nerve

Técnica mínimamente invasiva para la inyección en nervio óptico de la rata

The rat optic nerve is a useful model for stem cell regeneration research. Direct injection into the rat optic nerve allows delivery into the central ...

Visual Evoked Potential Recording in a Rat Model of Experimental Optic Nerve Demyelination

Grabación de potencial evocado visual en un modelo de rata de Experimental del Nervio Óptico Desmielinización

The visual evoked potential (VEP) recording is widely used in clinical practice to assess the severity of optic neuritis in its acute phase, and to ...

Exposure of the Pig CNS for Histological Analysis: A Manual for Decapitation, Skull Opening, and Brain Removal

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Pigs have become increasingly popular in large-animal translational neuroscience research as an economically and ethically feasible substitute to ...