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.

1. Prepare artificial cerebrospinal fluid (aCSF)1. We first prepare a 2L of a 10X stock of aCSF without magnesium or calcium. Reagents listed in millimolar. Catalog numbers refer to Sigma/Aldrich.
<table border="1" cellpadding="0">
<tbody>
<tr><td colspan="4">2 Liters 10X aCSF (without Mg or Ca)</td></tr>
<tr><td>Reagent</td><td>mM</td><td>g/2L</td><td>Catalog</td></tr>
<tr><td>KCl</td><td>40</td><td>5.96</td><td>P-9333</td></tr>
<tr><td>NaCl</td><td>1280</td><td>149...

<ol>
	<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>Goulding, M., Pfaff, S. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+circuits+that+generate+simple+rhythmic+behaviors+in+vertebrates.">Development of circuits that generate simple rhythmic behaviors in vertebrates.</a> Curr Opin Neurobiol. 15 (1), 14-20 (2005).</li><li>Gallarda, B. <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, (2008).</li><li>Gosgnach, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=V1+spinal+neurons+regulate+the+speed+of+vertebrate+locomotor+outputs.">V1 spinal neurons regulate the speed of vertebrate locomotor outputs.</a> Nature. 440 (7081), 215-219 (2006).</li><li>Lanuza, G. M., Gosgnach, S., Pierani, A., Jessell, T. M., 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=Genetic+identification+of+spinal+interneurons+that+coordinate+left-right+locomotor+activity+necessary+for+walking+movements.">Genetic identification of spinal interneurons that coordinate left-right locomotor activity necessary for walking movements.</a> Neuron. 42 (3), 375-386 (2004).</li><li>Jiang, Z., Carlin, K. P., Brownstone, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+in+vitro+functionally+mature+mouse+spinal+cord+preparation+for+the+study+of+spinal+motor+networks.">An in vitro functionally mature mouse spinal cord preparation for the study of spinal motor networks.</a> Brain Res. 816 (2), 493-499 (1999).</li><li>Ziskind-Conhaim, L., Gao, B. X., Hinckley, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ethanol+dual+modulatory+actions+on+spontaneous+postsynaptic+currents+in+spinal+motoneurons.">Ethanol dual modulatory actions on spontaneous postsynaptic currents in spinal motoneurons.</a> J Neurophysiol. 89 (2), 806-813 (2003).</li><li>Tabak, J., Rinzel, J., O'Donovan, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+activity-dependent+network+depression+in+the+expression+and+self-regulation+of+spontaneous+activity+in+the+developing+spinal+cord.">The role of activity-dependent network depression in the expression and self-regulation of spontaneous activity in the developing spinal cord.</a> J Neurosci. 21 (22), 8966-8978 (2001).</li><li>Chub, N., Mentis, G. Z., O'Donovan, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chloride-sensitive+MEQ+fluorescence+in+chick+embryo+motoneurons+following+manipulations+of+chloride+and+during+spontaneous+network+activity.">Chloride-sensitive MEQ fluorescence in chick embryo motoneurons following manipulations of chloride and during spontaneous network activity.</a> J Neurophysiol. 95 (1), 323-330 (2006).</li></ol>

The isolated neonatal spinal cord provides a tractable method of studying nervous system development1,2. Within the lumbar spinal cord of neonatal rodents, central pattern generating circuitry can produce fictive locomotion in the presence of neurotransmitters. This fictive locomotion consists of rhythmic increases in activity, bursts, that are produced at 0.2 to 0.5 Hz. These bursts are organized to produce left-right alternation along the length of the cord and flexor-extensor alternation (ipsilateral L2 relative to L5...

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		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>KCl</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>P-9333</td>
<td>&nbsp;</td>
<tr>
<td>NaCl	</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>S-7653</td>
<td>&nbsp;</td>
<tr>
<td>NaHCO3</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>S-6297</td>
<td>&nbsp;</td>
<tr>
<td>NaH2PO4</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>S-9638</td>
<td>&nbsp;</td>
<tr>
<td>Glucose</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>D-9434</td>
<td>&nbsp;</td>
<tr>
<td>CaCl2</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>C-5080</td>
<td>&nbsp;</td>
<tr>
<td>MgSO4</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>M-5921</td>
<td>&nbsp;</td>
<tr>
<td>Large Scissors</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>14070-12</td>
<td>&nbsp;</td>
<tr>
<td>Forceps</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>11050-10</td>
<td>&nbsp;</td>
<tr>
<td>Fine Scissors</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>15000-10</td>
<td>&nbsp;</td>
<tr>
<td>Insect pins</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>26002-10</td>
<td>&nbsp;</td>
<tr>
<td>Sylgard 184 (Dow-Corning)</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>1L volumetric flask</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>100mL volumetric flask</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>

spinal cord electrophysiology

The neonatal mouse spinal cord is a model for studying the development of neural circuitries and locomotor movement. We demonstrate the spinal cord dissection and preparation of recording bath artificial cerebrospinal fluid used for locomotor studies. Once dissected, the spinal cord ventral nerve roots can be attached to a recording electrode to record the electrophysiologic signals of the central pattern generating circuitry within the lumbar cord.

Watch this Scientific Journal Video about Spinal Cord Electrophysiology at JoVE.com

Spinal Cord Electrophysiology

A demonstration of the isolation of neonatal mouse spinal cord for electrophysiologic studies.

The neonatal mouse spinal cord is a model for studying the development of neural circuitries and locomotor movement. We demonstrate the spinal cord ...

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21.0K vistas.  Howard Hughes Medical Institute and Gene Expression Laboratory. Una demostración del aislamiento de la médula espinal de ratón neonatal para los estudios electrofisiológicos.