Name: spinal cord electrophysiology ii: extracellular suction electrode fabrication
Uploaded: 2011-02-20T23:59:00
Description: Watch this Scientific Journal Video about Spinal Cord Electrophysiology II: Extracellular Suction Electrode Fabrication at JoVE.com

Samuel L. Pfaff is a Professor in the Gene Expression Laboratories at The Salk Institute for biological Studies and an Investigator in the Howard Hughes Medical Institute. This work was supported by the Christopher and Dana Reeve Foundation. Joe Belcovson, Kent Schnoeker and Mike Sullivan in Multimedia Resources at the Salk Institute provided assistance with photography and editing.

Electrophysiological recordings of isolated spinal cords can reveal genetic and developmental changes to neural circuitry 1. We previously demonstrated a method to dissect neonatal mouse spinal cords 2. Here we present a method to prepare suction electrodes useful in recording fictive locomotion in isolated spinal cords 3.
The tip of a plastic tubing electrode can be drawn to a very fine tip by hand using a low-temperature alcohol lamp 3-5. The plas...

<ol>
	<li>Gallarda, B. W., Sharpee, T. O., Pfaff, S. L., Alaynick, W. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Defining+rhythmic+locomotor+burst+patterns+using+a+continuous+wavelet+transform.">Defining rhythmic locomotor burst patterns using a continuous wavelet transform.</a> Ann N Y Acad Sci. 1198, 133-139 (2010).</li><li>Meyer, A., Gallarda, B. W., Pfaff, S., Alaynick, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spinal+cord+electrophysiology.">Spinal cord electrophysiology.</a> J Vis Exp. , (2010).</li><li>Gallarda, B. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Segregation+of+axial+motor+and+sensory+pathways+via+heterotypic+trans-axonal+signaling.">Segregation of axial motor and sensory pathways via heterotypic trans-axonal signaling.</a> Science. 320, 233-236 (2008).</li><li>Landmesser, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+development+of+motor+projection+patterns+in+the+chick+hind+limb.">The development of motor projection patterns in the chick hind limb.</a> J Physiol. 284, 391-414 (1978).</li><li>Myers, C. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cholinergic+input+is+required+during+embryonic+development+to+mediate+proper+assembly+of+spinal+locomotor+circuits.">Cholinergic input is required during embryonic development to mediate proper assembly of spinal locomotor circuits.</a> Neuron. 46, 37-49 (2005).</li><li>Chanin, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+determination+of+chloride+by+use+of+the+silver-silver+chloride+electrode.">The determination of chloride by use of the silver-silver chloride electrode.</a> Science. 119, 323-324 (1954).</li><li>Goulding, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Circuits+controlling+vertebrate+locomotion:+moving+in+a+new+direction.">Circuits controlling vertebrate locomotion: moving in a new direction.</a> Nat Rev Neurosci. 10, 507-518 (2009).</li></ol>

Nervous system development can be studied using isolated rodent spinal cords. In the presence of neurotransmitters, fictive locomotion can be generated from the spinal cord in the form of patterned electrical activity 1,3. These rhythmic bursts are produced at 0.2 to 0.5 Hz and are patterned in left-right and flexor-extensor alternations. At different developmental stages, the robustness and patterns of this activity varies 1. Genetic mutations can also disturb the patterning of this activity 3...

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<td>PTFE Sub Lite Wall Tubing (Small tubing)</td>
<td>&nbsp;</td>
<td>Zeus</td>
<td>36AWG</td>
<td>0.005&#x201D;ID x 0.003&#x201D; Wall (Small Parts) Also available in 0.003&#x201D; to 0.006&#x201D;</td>
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<td>Large tubing (0.86mm (0.34&#x201D;))</td>
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<td>Clay Adams Brand Intramedic Becton Dickinson and Company</td>
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<td>0.86mm (0.34&#x201D;) O.D. 1.27mm (.050&#x201D;)</td>
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<td>Electrode Barrel</td>
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<td>A-M Systems</td>
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<td>Adhesive: Silicone caulk</td>
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<td>Bleach</td>
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<td>Xylene</td>
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<td>Silver wire: 0.010&#x201D;</td>
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<td>A-M Systems</td>
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<td>Insect pins: Austerlitz 0.1mm</td>
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<td>Fine Science Tools</td>
<td>26002-10</td>
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<td>Magnetic Stand</td>
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<td>Narishige</td>
<td>GJ-8</td>
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<td>Micromanipulator</td>
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<td>Bipolar Portable Physiodata Amplifier System</td>
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<td>Grass Industries</td>
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<td>ANALOG TO DIGITAL CARD</td>
<td>&nbsp;</td>
<td>National Instruments</td>
<td>6035E</td>
<td>&nbsp;</td>
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<td>Air Table; Vibraplane</td>
<td>&nbsp;</td>
<td>Kinetic Systems</td>
<td>&nbsp;</td>
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spinal cord electrophysiology ii: extracellular suction electrode fabrication

Development of neural circuitries and locomotion can be studied using neonatal rodent spinal cord central pattern generator (CPG) behavior. We demonstrate a method to fabricate suction electrodes that are used to examine CPG activity, or fictive locomotion, in dissected rodent spinal cords. The rodent spinal cords are placed in artificial cerebrospinal fluid and the ventral roots are drawn into the suction electrode. The electrode is constructed by modifying a commercially available suction electrode. A heavier silver wire is used instead of the standard wire given by the commercially available electrode. The glass tip on the commercial electrode is replaced with a plastic tip for increased durability. We prepare hand drawn electrodes and electrodes made from specific sizes of tubing, allowing consistency and reproducibility. Data is collected using an amplifier and neurogram acquisition software. Recordings are performed on an air table within a Faraday cage to prevent mechanical and electrical interference, respectively.

Watch this Scientific Journal Video about Spinal Cord Electrophysiology II: Extracellular Suction Electrode Fabrication at JoVE.com

Spinal Cord Electrophysiology II: Extracellular Suction Electrode Fabrication

A demonstration of the fabrication and use of an extracellular suction electrode used to measure electrophysiological recordings of neonatal rodent spinal cords in vitro

Development of neural circuitries and locomotion can be studied using neonatal rodent spinal cord central pattern generator (CPG) behavior. We demonstrate ...

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