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; Questo lavoro è stato sostenuto dal programma intramurale del National Institute of Neurological Disorder and Stroke dei National Institutes of Health. Vorremmo anche ringraziare il Dr. George Mentis per i suoi contributi precedenti al metodo e ai dati in Figura 3.</p

Destrani fluorescenti sono stati utilizzati come strumenti anatomici e per l&#39;imaging attività neuronale 1-4. Campi et al. (2009) 4 pubblicato un protocollo per l&#39;applicazione di ioni e della tensione di coloranti sensibili ai tratti assonali con un focus sul midollo spinale come sistema modello. Qui si descrive una procedura più dettagliata per l&#39;applicazione colorante fluorescente e / o ione-sensibile alle radici taglio ventrali, radici dorsali o qualsiasi tratto neuronale d...

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	<li>Nance, D. M., Burns, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fluorescent+dextrans+as+sensitive+anterograde+neuroanatomical+tracers:+applications+and+pitfalls.">Fluorescent dextrans as sensitive anterograde neuroanatomical tracers: applications and pitfalls.</a> Brain Res. Bull. 25, 139-145 (1990).</li><li>Glover, J. C., Petursdottir, G., Jansen, J. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fluorescent+dextran-amines+used+as+axonal+tracers+in+the+nervous+system+of+the+chicken+embryo.">Fluorescent dextran-amines used as axonal tracers in the nervous system of the chicken embryo.</a> J. Neurosci. Methods. 18, 243-254 (1986).</li><li>McPherson, D. R., McClellan, A. D., O'Donovan, M. 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J., Ho, S., Sholomenko, G., Yee, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Real-time+imaging+of+neurons+retrogradely+and+anterogradely+labelled+with+calcium-sensitive+dyes.">Real-time imaging of neurons retrogradely and anterogradely labelled with calcium-sensitive dyes.</a> J. Neurosci. Methods. 46, 91-106 (1993).</li><li>Wenner, P., Tsau, Y., Cohen, L. B., O'Donovan, M. J., Dan, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Voltage-sensitive+dye+recording+using+retrogradely+transported+dye+in+the+chicken+spinal+cord:+staining+and+signal+characteristics.">Voltage-sensitive dye recording using retrogradely transported dye in the chicken spinal cord: staining and signal characteristics.</a> J. Neurosci. 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Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+functional+impairment+of+sensory-motor+connectivity+in+a+mouse+model+of+spinal+muscular+atrophy.">Early functional impairment of sensory-motor connectivity in a mouse model of spinal muscular atrophy.</a> Neuron. 69, 453-467 (2011).</li><li>Blivis, D., Mentis, Z., O'Donovan, J. M. G., Lev-Tov, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Studies+of+sacral+neurons+involved+in+activation+of+the+lumbar+central+pattern+generator+for+locomotion+in+the+neonatal+rodent+spinal+cord.">Studies of sacral neurons involved in activation of the lumbar central pattern generator for locomotion in the neonatal rodent spinal cord.</a> Soc. Neurosci. Abstr. 564.8, (2009).</li><li>O'Donovan, M. 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<p class="jove_content"&gt; Descriviamo qui un protocollo semplice e conveniente per l&#39;introduzione di coloranti in neuroni, nervi e tratti della colonna vertebrale. Questo metodo consiste nell&#39;esporre i percorsi identificati anatomiche a una soluzione altamente concentrata colorante per la durata del processo di caricamento. Ciò si traduce in etichettatura retrogrado del sito bersaglio con sfondo poco rispetto microiniezioni, applicazione bagno e tecniche di elettroporazione. Tuttavia, il metodo è limitato ai siti nel sistema nervoso...

<table border="1"&gt;<tbody&gt;<tr&gt;<td colspan="2"<strong&gt; Nome del materiale</strong</td&gt;<td<strong&gt; Azienda</strong</td&gt;<td<strong&gt; Numero di catalogo</strong</td&gt;<td<strong&gt; Commenti</strong</td&gt;</tr&gt;<tr&gt;<td rowspan="2"&gt; Tubi</td&gt;<td&gt; PE90 (IDxOD: 0,034 &quot;x0.050&quot;; Spessore parete: 0.008 &quot;)</td&gt;<td&gt; Clay Adams Marca Intermedic</td&gt;<td&gt; 427.421</td&gt;<td&gt; Per Tipo I e Tipo II pipette</td&gt;</tr&gt;<tr&gt;<td&gt; NSF-51 (IDxOD: 1/16x1/8; Spessore parete: 1/32)</td&gt;<td&gt; PharMed BPT, Cole-Parmer</td&gt;<td&gt; AY242002</td&gt;<td&gt; Per Tubi flessibili</td&gt;</tr&gt;<tr&gt;<td&gt; Siringa</td&gt;<td&gt; Siringa da insulina da 1 ml U-100</td&gt;<td&gt; Becton Dickinson</td&gt;<td&gt; 329.650</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Needle</td&gt;<td&gt; 19G x1-1/2 &quot;Metal Hub Needle</td&gt;<td&gt; MONOJECT</td&gt;<td&gt; 200.136</td&gt;<td&gt; Collegamento di tipo II pipetta siringa</td&gt;</tr&gt;<tr&gt;<td&gt;</td&gt;<td&gt; Alcohol Lamp</td&gt;<td&gt;</td&gt;<td&gt;</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Rubinetto</td&gt;<td&gt; Valvola a tre vie con Maschio Adattatore Luer slip</td&gt;<td&gt; Baxter Healthcare Corp.</td&gt;<td&gt; 2C6241</td&gt;<td&gt; Usa con siringa</td&gt;</tr&gt;<tr&gt;<td rowspan="3"&gt; Titolare</td&gt;<td&gt; Supporto H-1 elettrodo</td&gt;<td&gt; Narishige</td&gt;<td&gt; H-1/12</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Magnet supporto</td&gt;<td&gt; Narishige</td&gt;<td&gt;-8 GJ</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Micromanipolatore</td&gt;<td&gt; Narishige</td&gt;<td&gt; M-3333</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Pinzetta</td&gt;<td&gt; S &amp; T DUMONT svizzera 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; * Questi sono materiali suggeriti. Può essere sostituito con qualsiasi prodotti compatibili</p

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

Etichettatura retrograda di neuroni è uno standard 1,2 metodo anatomica che è stata anche utilizzata per caricare e calcio voltaggio-coloranti sensibili in neuroni 3-6. Generalmente, i coloranti vengono applicate come cristalli solidi o mediante iniezione a pressione locale utilizzando pipette di vetro. Tuttavia, ciò può causare diluizione della intensità etichettatura colorante e ridotta, in particolare quando sono necessarie diverse ore per diffusione colorante. Qui mostriamo una tecnica semplice e di basso costo per l&#39;introduzione coloranti fluorescenti e ioni sensibile in neuroni con una pipetta di aspirazione polietilene riempito con la soluzione colorante. Questo metodo offre un modo affidabile per mantenere un&#39;elevata concentrazione del colorante in contatto con assoni tutta la procedura di caricamento.

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

Descriviamo una tecnica semplice ea basso costo per introdurre alta concentrazione di coloranti fluorescenti e calcio-sensibili in neuroni o qualsiasi tratto neuronale mediante una pipetta di aspirazione polietilene.

Caricamento in corso retrogrado di Nervi, tratti e radici spinali con le tinture fluorescenti

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. ...

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

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