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"> Lösningar </p><ul><li> Intracellulär lösning för ganglion celler (i mm): 100 K-glukonat, 8 KCl, 1 MgCl <sub>2,</sub> 1 EGTA, 10 HEPES, 4 ATP, 0,5 GTP, 90 mikroM Sulforhodamine B (för färgning), justerat till pH = 7,4 med KOH </li><li> Intracellulär lösning för bipolär celler (i mm): 90 K-glukonat, 8 KCl, 1 MgCl <sub>2,</sub> 10 BAPTA, 10 HEPES, 4 ATP, 0,5 GTP, 90 mikroM Sulforhodamine B (för färgning), justerat till pH = 7,4 med KOH </li><li> Extracellulär Ringers lösning (i mm): 112 NaCl, 5 glukos, 5 HEPES,...

<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> Fördelar: </p><ol><li> Alla celltyper är tillgängliga <br /></li><li> Enkel identifiering av celltyper, särskilt om en fluorescerande färg läggs till elektroden lösningen <br /></li><li> Farmakologiska agenter kan lätt nå målceller <br /></li><li> Plåstret clamp tekniken tillåter att undersöka betydelsen av olika jonkanaler i retina beräkningar. Samma information kan inte erhållas genom extracellulära spik inspelningar eller inspelningar med skarpa elektroder. <br /></li><li> Denna teknik kan tillämpas på andra djurmodelle...

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		<th>Type</th>
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		<th>Catalogue Number</th>
		<th>Comment</th>
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<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>
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<td>8-channel perfusion system</td>
<td>Microperfusion</td>
<td>Parker Hannfin corporation</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
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<td>Bits++</td>
<td>Digital Video Processor</td>
<td>Cambridge Research Systems</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
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<td>Infrared Oculars</td>
<td>Other</td>
<td>ITT</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
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<td>Adhesive silicone wells</td>
<td>Other</td>
<td>Molecular Probes</td>
<td>&nbsp;</td>
<td>20mm diameter, 1.0mm deep</td>
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<td>Membrane Filters</td>
<td>Filter</td>
<td>Milipore</td>
<td>HAWPO1300</td>
<td>.45&#181;m HA</td>
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<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>
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<td>IR camera</td>
<td>Micropscope mounted camera</td>
<td>Sony</td>
<td>SPT-M324</td>
<td>Extrawave HAD, B7W video camera</td>
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<td>Picrotoxin</td>
<td>Reagent</td>
<td>Sigma</td>
<td>P1675</td>
<td>&nbsp;</td>
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<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>
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whole-cell recordings of light evoked excitatory synaptic currents in the retinal slice

<p class="jove_content">Vi använder helcells-tekniken patch clamp att studera de synaptiska kretsar som ligger bakom visuell information bearbetning i näthinnan. I den här videon kommer vi att guida dig genom processen att utföra helcells-inspelningar av ljuset framkallade strömmar av enskilda celler i näthinnan skiva beredning. Vi använder vattenlevande tiger salamandern som en djurmodell. Vi börjar med att beskriva dissekering av ögat och visa hur skivor är monterade för elektrofysiologiska inspelningar. När skiva är placerad i inspelningen kammare, visar vi hur du utför helcells-inspelningar spänning klämma. Vi projicerar då visuella stimuli på fotoreceptorer i segmentet att framkalla ljus framkallat Dagens svar. Under inspelningen vi BEGJUTA segmentet med farmakologiska medel, där en 8-kanals perfusion systemet tillåter oss att snabbt växla mellan olika aktörer. Näthinnans skiva preparatet används ofta för inspelningar patch clamp i näthinnan, i synnerhet för att studera amakrina eller bipolär celler, som inte är tillgängliga i hela montera beredning.</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">Denna video visar processen helcells-inspelningar spänning klämma i retinal bit av vattenlevande tiger salamandern. Vi visar förberedelserna för den del, liksom hur man utför inspelningar patch clamp under visuell stimulering av näthinnan.</p>

Helcells-Inspelningar av Ljus Evoked excitatoriska Synaptic strömmar i näthinnan 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 ...

Research

JoVE Journal

Biology

Horizontal Slice Preparation of the Retina

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In this video, we demonstrate the procedure for isolating whole brains from adult Drosophila in preparation for recording from single neurons. We begin by ...

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The light microscope is a basic tool for the cell biologist, who should have a thorough understanding of how it works, how it should be aligned for ...

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Rod and cone photoreceptors in the retina are responsible for light detection. In darkness, cyclic nucleotide-gated (CNG) channels in the outer segment ...

Whole-Cell Recording of Calcium Release-Activated Calcium (CRAC) Currents in Human T Lymphocytes

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Explant culture allows manipulation of developing organs at specific time points&#160;and is therefore an important method for the developmental ...

<em>In vivo</em> Clonal Tracking of Hematopoietic Stem and Progenitor Cells Marked by Five Fluorescent Proteins using Confocal and Multiphoton Microscopy

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<em>In Vivo</em> 4-Dimensional Spårning av Hematopoietic Stem och stamceller i vuxen mus calvarial Benmärg

Through a delicate balance between quiescence and proliferation, self renewal and production of differentiated progeny, hematopoietic stem cells (HSCs) ...

Method for the Assessment of Effects of a Range of Wavelengths and Intensities of Red/near-infrared Light Therapy on Oxidative Stress <em>In Vitro</em>

Method for the Assessment of Effects of a Range of Wavelengths and Intensities of Red/near-infrared Light Therapy on Oxidative Stress In Vitro

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Red/near-infrared light therapy (R/NIR-LT), delivered by laser or light emitting diode (LED), improves functional and morphological outcomes in a range of ...