Name: whole-cell recordings of light evoked excitatory synaptic currents in the retinal slice
Uploaded: 2008-07-02T15:50:00
Description: Watch this Scientific Journal Video about Whole-cell Recordings of Light Evoked Excitatory Synaptic Currents in the Retinal Slice at JoVE.com

<p class="jove_title"> Solutions </p><ul><li> Solution intracellulaire des cellules ganglionnaires (en mm): 100 K-gluconate, 8 KCl, MgCl 1 <sub>2,</sub> 1 EGTA, HEPES 10, 4 ATP, GTP 0,5, 90 uM Sulforhodamine B (pour la coloration), ajusté à pH = 7,4 avec KOH </li><li> Solution intracellulaire des cellules bipolaires (en mm): 90 K-gluconate, 8 KCl, MgCl <sub>2</sub> 1, 10 BAPTA, HEPES 10, 4 ATP, GTP 0,5, 90 uM Sulforhodamine B (pour la coloration), ajusté à pH = 7,4 avec KOH </li><li> Ringer extracellulaire de solution (en mM): 112 NaCl, glu...

<ol>
	<li>Cook, P. B., Lukasiewicz, P. D., McReynnolds, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=GABA(C)+receptors+control+adaptive+changes+in+a+glycinergic+inhibitory+pathway+in+salamander+retina.">GABA(C) receptors control adaptive changes in a glycinergic inhibitory pathway in salamander retina.</a> <em style='font-style: italic'>Journal of Neuroscience</em>. <b>20</b>, 806-812 (2000).</li><li>Heflin, S. J., Cook, P. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Narrow+and+wide+field+amacrine+cells+fire+action+potentials+in+response+to+depolarization+and+light+stimulation.">Narrow and wide field amacrine cells fire action potentials in response to depolarization and light stimulation.</a> <em style='font-style: italic'>Visual Neuroscience</em>. <b>24</b>, 197-206 (2007).</li><li>Ichinose, T., Shields, C. R., Lukasiewicz, 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=Sodium+Channels+in+the+Transient+Retinal+Bipolar+Cells+Enhance+Visual+Responses+in+Ganglion+Cells.">Sodium Channels in the Transient Retinal Bipolar Cells Enhance Visual Responses in Ganglion Cells.</a> <em style='font-style: italic'>J Neurosci.</em>. <b>25</b>, 1856-1865 (2005).</li></ol>

<p> Avantages: </p><ol><li> Tous les types de cellules sont accessibles <br /></li><li> Identification aisée des différents types cellulaires, en particulier si un colorant fluorescent est ajouté à la solution d&#39;électrode <br /></li><li> Les agents pharmacologiques peuvent facilement atteindre les cellules cibles <br /></li><li> La technique de patch-clamp permet d&#39;étudier le rôle des canaux ioniques différentes dans les calculs de la rétine. La même information ne peut être obtenu grâce à des enregistrements ou des enregis...

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		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Multiclamp 700A</td>
<td>Patch Clamp Amplifier</td>
<td>Molecular Devices</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>SZX9</td>
<td>Dissection Micropscope</td>
<td>Olympus</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>BX50WI</td>
<td>Upright Microscope</td>
<td>Olympus</td>
<td>&nbsp;</td>
<td>Equipped with 40x, 10x water immersion objective, fluorescent filters and mercury lamp</td>
</tr>
<tr>
<td>P-97</td>
<td>Micropipette Puller</td>
<td>Sutter Instruments</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Lucivid</td>
<td>Microscope image injector</td>
<td>Mbf bioscience</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>8-channel perfusion system</td>
<td>Microperfusion</td>
<td>Parker Hannfin corporation</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Bits++</td>
<td>Digital Video Processor</td>
<td>Cambridge Research Systems</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Infrared Oculars</td>
<td>Other</td>
<td>ITT</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Adhesive silicone wells</td>
<td>Other</td>
<td>Molecular Probes</td>
<td>&nbsp;</td>
<td>20mm diameter, 1.0mm deep</td>
</tr>
<tr>
<td>Membrane Filters</td>
<td>Filter</td>
<td>Milipore</td>
<td>HAWPO1300</td>
<td>.45&#181;m HA</td>
</tr>
<tr>
<td>Borosilicate Glass Capillaries</td>
<td>Electrode glass</td>
<td>World Precision Instruments</td>
<td>1B150-4</td>
<td>&nbsp;</td>
<tr>
<td>Syringe Filters</td>
<td>Filter</td>
<td>Whatman</td>
<td>6789-0402</td>
<td>4mm filters, .2um Nylon Membrane, Polypropylene housing</td>
</tr>
<tr>
<td>IR camera</td>
<td>Micropscope mounted camera</td>
<td>Sony</td>
<td>SPT-M324</td>
<td>Extrawave HAD, B7W video camera</td>
</tr>
<tr>
<td>Picrotoxin</td>
<td>Reagent</td>
<td>Sigma</td>
<td>P1675</td>
<td>&nbsp;</td>
<tr>
<td>Strychnine</td>
<td>Reagent</td>
<td>Sigma</td>
<td>S0532</td>
<td>&nbsp;</td>
<tr>
<td>Imidazole-4-acetic acid sodium salt</td>
<td>Reagent</td>
<td>Sigma</td>
<td>I7013</td>
<td>&nbsp;</td>
<tr>
<td>Larval tiger salamanders</td>
<td>Animal</td>
<td>Charles E. Sullivan (Nashville, TN)</td>
<td>&nbsp;</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>
	

whole-cell recordings of light evoked excitatory synaptic currents in the retinal slice

<p class="jove_content">Nous utilisons la technique de cellules entières patch clamp pour étudier les circuits synaptiques qui sous-tend le traitement des informations visuelles de la rétine. Dans cette vidéo, nous allons vous guider à travers le processus d&#39;exécution de cellules entières d&#39;enregistrements de la lumière des courants évoqués des cellules individuelles dans la préparation de tranches de la rétine. Nous utilisons la salamandre tigrée aquatiques comme un modèle animal. Nous commençons par décrire la dissection de l&#39;oeil et de montrer comment les tranches sont montés pour des enregistrements électrophysiologiques. Une fois que la tranche est placée dans la chambre d&#39;enregistrement, nous montrent comment effectuer la cellule entière enregistrements voltage. Nous avons ensuite projet de stimuli visuels sur les photorécepteurs de la tranche à susciter la lumière évoque les réponses actuelles. Pendant l&#39;enregistrement, nous perfuser la tranche avec des agents pharmacologiques, par lequel un système de perfusion à 8 canaux nous permet de basculer rapidement entre les différents agents. La préparation de tranches rétiniennes est largement utilisé pour les enregistrements de patch-clamp dans la rétine, en particulier d&#39;étudier ou de cellules amacrines bipolaire, qui ne sont pas accessibles dans une préparation toute monture.</p>

Watch this Scientific Journal Video about Whole-cell Recordings of Light Evoked Excitatory Synaptic Currents in the Retinal Slice at JoVE.com

Whole-cell Recordings of Light Evoked Excitatory Synaptic Currents in the Retinal Slice

<p class="jove_content">Cette vidéo montre le processus de la cellule entière enregistrements pince de tension dans la tranche la rétine de la salamandre aquatique tigre. Nous démontrons la préparation de la tranche ainsi que la façon d&#39;effectuer des enregistrements de patch-clamp pendant la stimulation visuelle de la rétine.</p>

Cellules entières Recordings of Light évoqués courants synaptiques excitateurs dans la tranche la rétine

We use the whole-cell patch clamp technique to study the synaptic circuitry that underlies visual information processing in the retina. In this video, we ...

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Horizontal Slice Preparation of the Retina

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Méthode d&#39;évaluation des effets d&#39;une gamme de longueurs d&#39;onde et des intensités de Luminothérapie Rouge / proche infrarouge sur le stress oxydatif<em&gt; In Vitro</em

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