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"> Oplossingen </p><ul><li> Intracellulaire oplossing voor het ganglion cellen (in mm): 100 K-gluconaat, 8 KCl, 1 MgCl <sub>2,</sub> 1 EGTA, 10 HEPES, 4 ATP, GTP 0.5, 90 uM sulforhodamine B (voor de kleuring), aangepast aan pH = 7,4 met KOH </li><li> Intracellulaire oplossing voor bipolaire cellen (in mm): 90 K-gluconaat, 8 KCl, 1 MgCl <sub>2,</sub> 10 BAPTA, 10 HEPES, 4 ATP, GTP 0.5, 90 uM sulforhodamine B (voor de kleuring), aangepast aan pH = 7,4 met KOH </li><li> Extracellulaire Ringer-oplossing (in mm): 112 NaCl, 5 Gluc...

<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> Voordelen: </p><ol><li> Alle celtypen zijn toegankelijk <br /></li><li> Eenvoudige identificatie van celtypen, met name wanneer een fluorescerende kleurstof wordt toegevoegd aan de elektrode oplossing <br /></li><li> Farmacologische middelen gemakkelijk kunt bereiken doelcellen <br /></li><li> De patch clamp techniek maakt het mogelijk om de rol van verschillende ionenkanalen te onderzoeken in het netvlies berekeningen. Dezelfde informatie kan niet worden verkregen via extracellulaire spike-opnamen of opnamen met scherpe elektroden. <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">We gebruiken de whole-cell patch clamp techniek om de synaptische circuits die ten grondslag ligt visuele informatieverwerking in het netvlies te bestuderen. In deze video, zullen wij begeleiden u door het proces van het uitvoeren van whole-cell-opnamen van het licht opgeroepen stromingen van de individuele cellen in het netvlies slice voorbereiding. We gebruiken de in het water levende tijger salamander als een diermodel. We beginnen met een beschrijving van de dissectie van het oog en laten zien hoe schijfjes zijn gemonteerd voor elektrofysiologische opnames. Zodra het schijfje is geplaatst in de opname kamer, we laten zien hoe je whole-cell voltage clamp opnames uit te voeren. Vervolgens hebben we het project visuele prikkels op de fotoreceptoren in de slice aan het licht opgewekte stroom reacties uitlokken. Tijdens de opname hebben we perfuseren het segment met farmacologische stoffen, waarbij een 8-kanaals perfusie-systeem stelt ons in staat om snel te schakelen tussen verschillende agenten. Het netvlies slice voorbereiding wordt veel gebruikt voor patch clamp opnames in het netvlies, met name om amacrine of bipolaire cellen, die niet toegankelijk zijn in een geheel-mount voorbereiding van studie.</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">Deze video toont het proces van het whole-cell voltage clamp opnamen in de retinale deel van de in het water levende tijger salamander. Tonen we de voorbereiding van het stuk, alsook hoe de patch clamp opnames uit te voeren tijdens visuele stimulatie van het netvlies.</p>

Whole-cell Opnames van Licht prikkelende synaptische Stromingen opgeroepen in het netvlies Slice

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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