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"> Soluções </p><ul><li> Solução intracelular de células ganglionares (em mM): 100 K gluconato, 8 KCl, 1 MgCl <sub>2,</sub> 1 EGTA, 10 HEPES, 4 ATP, GTP 0,5, 90 mM Sulforhodamine B (para coloração), ajustada para pH = 7,4 com KOH </li><li> Solução intracelular de células bipolares (em mM): 90 K gluconato, 8 KCl, 1 MgCl <sub>2,</sub> 10 BAPTA, 10 HEPES, 4 ATP, GTP 0,5, 90 mM Sulforhodamine B (para coloração), ajustada para pH = 7,4 com KOH </li><li> Solução de Ringer extracelular (em mM): 112 NaCl, Glicose 5, 5 ...

<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> Benefícios: </p><ol><li> Todos os tipos de células são acessíveis <br /></li><li> A fácil identificação de tipos de células, em particular se um corante fluorescente é adicionado à solução do eletrodo <br /></li><li> Agentes farmacológicos pode chegar facilmente a células-alvo <br /></li><li> A técnica de patch clamp permite investigar o papel dos canais iônicos em cálculos diferentes da retina. A mesma informação não pode ser obtida através de gravações extracelluar espiga ou gravações com eletrodos afiada. <br /><...

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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">Usamos a técnica de célula inteira patch clamp para estudar os circuitos sinápticos que está por trás de processamento de informações visuais na retina. Neste vídeo, que irá guiá-lo através do processo de realização de célula inteira gravações de luz correntes evocados de células individuais na preparação fatia da retina. Usamos o tigre aquáticos salamandra como modelo animal. Começamos por descrever a dissecação do olho e mostrar como as fatias são montados para as gravações eletrofisiológicas. Uma vez que a fatia é colocada na câmara de gravação, que demonstram como executar de célula inteira gravações braçadeira de tensão. Em seguida, projeto de estímulos visuais sobre os fotorreceptores na fatia de obter luz evocou respostas atual. Durante a gravação que perfundir a fatia com agentes farmacológicos, no qual um sistema de perfusão de 8 canais nos permite alternar rapidamente entre diferentes agentes. A preparação fatia da retina é amplamente usado para gravações de patch clamp na retina, em particular para estudar ou células amácrinas bipolar, que não são acessíveis em uma preparação de todo o monte.</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">Este vídeo mostra o processo de whole-cell recording voltagem clamp na fatia da retina do tigre aquáticos salamandra. Nós demonstramos a preparação da fatia, bem como a forma de realizar gravações patch clamp durante a estimulação visual da retina.</p>

De célula inteira Recordings of Light Evocados Excitatórios Correntes Synaptic na Slice Retinal

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 ...

Research

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