Name: retrograde loading of nerves, tracts, and spinal roots with fluorescent dyes
Uploaded: 2012-04-19T13:00:00
Description: Watch this Scientific Journal Video about Retrograde Loading of Nerves, Tracts, and Spinal Roots with Fluorescent Dyes at JoVE.com

<p class="jove_content"&gt; Este trabajo fue apoyado por el programa intramural de los Institutos Nacionales de Trastornos Neurológicos y Accidentes Cerebrovasculares de los Institutos Nacionales de Salud. También nos gustaría agradecer al Dr. George Mentis por sus contribuciones anteriores al método y los datos de la Figura 3.</p

Dextranos fluorescentes se han utilizado como herramientas anatómicas y para la actividad neuronal imagen 1-4. Campos et al, (2009) 4 publicó un protocolo para la aplicación de iones y sensibles al voltaje tintes de vías axonales con un enfoque en la médula espinal como un sistema modelo. Aquí se describe un procedimiento más detallado para la aplicación de tinte fluorescente y / o de iones sensibles a las raíces cortadas ventrales, raíces dorsales o cualquier tracto neuronal de ...

<ol>
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<p class="jove_content"&gt; Se describe aquí un protocolo simple y rentable para la introducción de colorantes en las neuronas, los nervios y tractos espinales. Este método consiste en exponer identificados vías anatómicas a una solución de tinte altamente concentrada para la duración del proceso de carga. Esto se traduce en el etiquetado retrógrada del sitio de destino con poca experiencia en comparación con microinyecciones, la aplicación de baños y técnicas de electroporación. Sin embargo, el método está limitado a los sitios ...

<table border="1"&gt;<tbody&gt;<tr&gt;<td colspan="2"<strong&gt; Nombre del Material</strong</td&gt;<td<strong&gt; Empresa</strong</td&gt;<td<strong&gt; Número de catálogo</strong</td&gt;<td<strong&gt; Comentarios</strong</td&gt;</tr&gt;<tr&gt;<td rowspan="2"&gt; Tubos</td&gt;<td&gt; PE90 (IDxOD: 0.034 &quot;x0.050&quot;, Espesor de pared: 0.008 &quot;)</td&gt;<td&gt; Clay Adams Marca Intermedic</td&gt;<td&gt; 427421</td&gt;<td&gt; Por tipo I y tipo II pipetas</td&gt;</tr&gt;<tr&gt;<td&gt; NSF-51 (IDxOD: 1/16x1/8; Espesor de pared: 1/32)</td&gt;<td&gt; PharMed BPT, Cole-Parmer</td&gt;<td&gt; AY242002</td&gt;<td&gt; Para los tubos flexibles</td&gt;</tr&gt;<tr&gt;<td&gt; Jeringa</td&gt;<td&gt; 1 ml jeringa de insulina U-100</td&gt;<td&gt; Becton Dickenson</td&gt;<td&gt; 329650</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Aguja</td&gt;<td&gt; 19G x1-1/2 &quot;Metal cubo de la aguja</td&gt;<td&gt; MONOJECT</td&gt;<td&gt; 200136</td&gt;<td&gt; Conexión de Tipo II pipeta a la jeringa</td&gt;</tr&gt;<tr&gt;<td&gt;</td&gt;<td&gt; El alcohol de la lámpara</td&gt;<td&gt;</td&gt;<td&gt;</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Llave de paso</td&gt;<td&gt; Llave de tres vías con adaptador Luer Slip Mujer</td&gt;<td&gt; Baxter Healthcare Corp.</td&gt;<td&gt; 2C6241</td&gt;<td&gt; Utilice con la jeringa</td&gt;</tr&gt;<tr&gt;<td rowspan="3"&gt; Soporte</td&gt;<td&gt; Titular de la H-1 electrodo</td&gt;<td&gt; Narishige</td&gt;<td&gt; H-1/12</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Imán soporte</td&gt;<td&gt; Narishige</td&gt;<td&gt; GJ-8</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Micromanipulador</td&gt;<td&gt; Narishige</td&gt;<td&gt; M-3333</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Pinzas</td&gt;<td&gt; S &amp; T DUMONT suiza 00632-11</td&gt;<td&gt; DUMONT</td&gt;<td&gt; JF-5 TC</td&gt;<td&gt;</td&gt;</tr&gt;</tbody&gt;</table&gt;<p class="jove_content"&gt; * Estos son los materiales sugeridos. Puede ser reemplazado por cualquiera de los productos compatibles</p

retrograde loading of nerves, tracts, and spinal roots with fluorescent dyes

Etiquetado retrógrada de las neuronas es un 1,2 anatómica método estándar que se ha utilizado también para cargar calcio y tintes sensibles al voltaje en neuronas 3-6. En general, los colorantes se aplican en forma de cristales sólidos o por inyección a presión local utilizando pipetas de vidrio. Sin embargo, esto puede resultar en la dilución de la intensidad colorante y etiquetado reducida, particularmente cuando se requieren varias horas para la difusión del colorante. Aquí se demuestra una técnica sencilla y de bajo costo para la introducción de colorantes fluorescentes y de iones sensibles a las neuronas con una pipeta de succión de polietileno llena de la solución de colorante. Este método ofrece una forma fiable para mantener una alta concentración del colorante en contacto con los axones en todo el procedimiento de carga.

Watch this Scientific Journal Video about Retrograde Loading of Nerves, Tracts, and Spinal Roots with Fluorescent Dyes at JoVE.com

Retrograde Loading of Nerves, Tracts, and Spinal Roots with Fluorescent Dyes

Se describe una técnica sencilla y de bajo costo para la introducción de alta concentración de colorantes fluorescentes y sensible de calcio en las neuronas o cualquier aparato neuronal con una pipeta de succión de polietileno.

Cargando retrógrada de los nervios, panfletos, y las raíces espinales con tintes fluorescentes

Retrograde labeling of neurons is a standard anatomical method1,2 that has also been used to load calcium and voltage-sensitive dyes into neurons3-6. ...

Research

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The fusion of synaptic vesicles with the plasma membrane (exocytosis) is a required step in neurotransmitter release and neuronal communication. The ...

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Drosophila melanogaster is emerging as a powerful model system for studying the development and function of the nervous system, particularly because of ...

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Voltage-sensitive dye imaging of neurons is a key methodology for the understanding of how neuronal networks are organised and how the simultaneous ...

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

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Marcaje fluorescente retrógrada Permite dirigida extracelular sola unidad de grabación de neuronas identificadas<em&gt; En vivo</em

The overall goal of this method is to record single-unit responses from an identified population of neurons. In vivo electrophysiological recordings from ...

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Neural stem cells (NSCs) can self-renew and differentiate into neurons and glia. Transplanted NSCs can replace lost neurons and glia after spinal cord ...

Retrograde Labeling of Retinal Ganglion Cells in Adult Zebrafish with Fluorescent Dyes

Etiquetado retrógrada de células ganglionares retinianas en adultos de pez cebra con tintes fluorescentes

As retrograde labeling retinal ganglion cells (RGCs) can isolate RGCs somata from dying sites, it has become the gold standard for counting RGCs in RGCs ...

In Vivo Electrophysiological Measurements on Mouse Sciatic Nerves

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En Vivo electrofisiológicos mediciones sobre los nervios ciáticos de ratones

Electrophysiological studies allow a rational classification of various neuromuscular diseases and are of help, together with neuropathological ...

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Studies in the mammalian neocortex have enabled unprecedented resolution of cortical structure, activity, and response to neurodegenerative insults by ...

Imaging Membrane Potential with Two Types of Genetically Encoded Fluorescent Voltage Sensors

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Genetically encoded voltage indicators (GEVIs) have improved to the point where they are beginning to be useful for in vivo recordings. While the ultimate ...

Retrograde Neuroanatomical Tracing of Phrenic Motor Neurons in Mice

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Phrenic motor neurons are cervical motor neurons originating from C3 to C6 levels in most mammalian species. Axonal projections converge into phrenic ...