Name: preparation and culture of chicken auditory brainstem slices
Uploaded: 2011-03-21T13:00:00
Description: Watch this Scientific Journal Video about Preparation and Culture of Chicken Auditory Brainstem Slices at JoVE.com

Actuels et anciens membres du laboratoire Rubel.

1. Préparation de la dissection Zone <ol><li> Continuellement bulle artificielle liquide céphalo-rachidien (ACSF), avec un mélange de 95% d&#39;O 2 / 5% de CO 2 (pH entre 7.2 à 7.4, l&#39;osmolarité de 295 à 310 mOsm / l). </li><li> Alors que l&#39;ACSF est en ébullition, nettoyer la zone de travail avec de l&#39;EtOH 70%. Aussi nettoyer les vibratome et lame de tranchage. Rincer la lame à l&#39;eau distillée (dH 2 O). </li><li> Endroit propre tampon d&#39;absorption de liquid...

<ol>
	<li>Howard, M. A., Burger, R. M., Rubel, E. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+developmental+switch+to+GABAergic+inhibition+dependent+on+increases+in+Kv1-type+K++currents.">A developmental switch to GABAergic inhibition dependent on increases in Kv1-type K+ currents.</a> J Neurosci. 27, 2112-2123 (2007).</li><li>Kuba, H., Koyano, K., Ohmori, H. <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+membrane+conductance+improves+coincidence+detection+in+the+nucleus+laminaris+of+the+chicken.">Development of membrane conductance improves coincidence detection in the nucleus laminaris of the chicken.</a> J Physiol. 540, 529-542 (2002).</li><li>Gao, H., Lu, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+development+of+intrinsic+and+synaptic+properties+of+chicken+nucleus+laminaris+neurons.">Early development of intrinsic and synaptic properties of chicken nucleus laminaris neurons.</a> Neuroscience. 153, 131-143 (2008).</li><li>Sanchez, J. T., Wang, Y., Rubel, E. W., Barria, A. <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+glutamatergic+synaptic+transmission+in+binaural+auditory+neurons.">Development of glutamatergic synaptic transmission in binaural auditory neurons.</a> J Neurophysiol. 104, 1774-1789 (2010).</li><li>Jackson, H., Hackett, J. T., Rubel, E. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Organization+and+development+of+brain+stem+auditory+nuclei+in+the+chick:+ontogeny+of+postsynaptic+responses.">Organization and development of brain stem auditory nuclei in the chick: ontogeny of postsynaptic responses.</a> J Comp Neurol. 210, 80-86 (1982).</li><li>Rubel, E. W., Parks, T. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Organization+and+development+of+brain+stem+auditory+nuclei+of+the+chicken:+tonotopic+organization+of+n.+magnocellularis+and+n.+laminaris.">Organization and development of brain stem auditory nuclei of the chicken: tonotopic organization of n. magnocellularis and n. laminaris.</a> J Comp Neurol. 164, 411-433 (1975).</li><li>Rubel, E. W., Smith, D. J., Miller, L. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Organization+and+development+of+brain+stem+auditory+nuclei+of+the+chicken:+ontogeny+of+n.+magnocellularis+and+n.+laminaris.">Organization and development of brain stem auditory nuclei of the chicken: ontogeny of n. magnocellularis and n. laminaris.</a> J Comp Neurol. 166, 469-489 (1976).</li><li>Stoppini, L., Buchs, P. A., Muller, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simple+method+for+organotypic+cultures+of+nervous+tissue.">A simple method for organotypic cultures of nervous tissue.</a> J Neurosci Methods. 37, 173-182 (1991).</li><li>Reyes, A. D., Rubel, E. W., Spain, W. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Membrane+properties+underlying+the+firing+of+neurons+in+the+avian+cochlear+nucleus.">Membrane properties underlying the firing of neurons in the avian cochlear nucleus.</a> J Neurosci. 14, 5352-5364 (1994).</li><li>Monsivais, P., Rubel, E. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accommodation+enhances+depolarizing+inhibition+in+central+neurons.">Accommodation enhances depolarizing inhibition in central neurons.</a> J Neurosci. 21, 7823-7830 (2001).</li><li>Lu, T., Trussell, L. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+and+elimination+of+endbulb+synapses+in+the+chick+cochlear+nucleus.">Development and elimination of endbulb synapses in the chick cochlear nucleus.</a> J Neurosci. 27, 808-8017 (2007).</li><li>Kuba, H., Yamada, R., Fukui, I., Ohmori, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tonotopic+specialization+of+auditory+coincidence+detection+in+nucleus+laminaris+of+the+chick.">Tonotopic specialization of auditory coincidence detection in nucleus laminaris of the chick.</a> J Neurosci. 25, 1924-1934 (2005).</li><li>Trussell, L. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cellular+mechanisms+for+preservation+of+timing+in+central+auditory+pathways.">Cellular mechanisms for preservation of timing in central auditory pathways.</a> Curr Opin Neurobiol. 7, 487-492 (1997).</li><li>Trussell, L. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synaptic+mechanisms+for+coding+timing+in+auditory+neurons.">Synaptic mechanisms for coding timing in auditory neurons.</a> Annu Rev Physiol. 61, 477-496 (1999).</li><li>Sorensen, S. A., Rubel, E. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+level+and+integrity+of+synaptic+input+regulates+dendrite+structure.">The level and integrity of synaptic input regulates dendrite structure.</a> J Neurosci. 26, 1539-1550 (2006).</li></ol>

Depuis plusieurs décennies, la préparation de tranches aiguë du tronc cérébral du poulet a été utilisée pour étudier le traitement auditif 9, 10. Cette approche a fourni une énorme quantité de données in vitro sur le traitement binaural physiologiques des deux états de développement et de maturité 4, 11, 12. On connaît bien ce circuit très spécialisé et le rôle de chaque noyau joue dans le traitement temporel des sons 13, 14. En fait, à court terme ...

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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>Chicken ACSF</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>NaCl</td>
<td>&nbsp;</td>
<td>Fisher Scientific</td>
<td>M-11624</td>
<td>130 mM</td>
</tr>
<tr>
<td>NaHCO3</td>
<td>&nbsp;</td>
<td>Fisher Scientific</td>
<td>M-10576</td>
<td>26 mM</td>
</tr>
<tr>
<td>KCl</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>P-9333</td>
<td>3 mM</td>
</tr>
<tr>
<td>NaH2PO4</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>S-8282</td>
<td>1.25 mM</td>
</tr>
<tr>
<td>Glucose</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>G-7528</td>
<td>10 mM</td>
</tr>
<tr>
<td>MgCl2</td>
<td>&nbsp;</td>
<td>Fisher Scientific</td>
<td>M33-500</td>
<td>1 mM</td>
</tr>
<tr>
<td>CaCl2</td>
<td>&nbsp;</td>
<td>Acros Organics</td>
<td>423525000</td>
<td>2 mM</td>
</tr>
<tr>
<td>Chicken culture medium (store at 4&deg;C)</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>advanced minimum essential medium (with NEAA, sodium pyruvate at 110 mg/l, without L-glutamate)</td>
<td>&nbsp;</td>
<td>Gibco Invitrogen</td>
<td>12492-013</td>
<td>48.00%</td>
</tr>
<tr>
<td>Earl's balanced salt solution</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>E-2888</td>
<td>24.00%</td>
</tr>
<tr>
<td>L-glutamine (200 mM)</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>G-7513</td>
<td>1.00%</td>
</tr>
<tr>
<td>Glucose solution (200 g/l, sterile filtered)</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>G-7528</td>
<td>2.75%</td>
</tr>
<tr>
<td>horse serum (heat inactivated, sterile filtered)</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>H-1138</td>
<td>24.00%</td>
</tr>
<tr>
<td>Penicillin-streptomycin*</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>P-0781</td>
<td>1.00 ml 
* = Add to 100 ml culture medium if needed to prevent contamination</td>
</tr>
<tr>
<td>Specific equipment</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Millicell-CM cell culture inserts</td>
<td>PICMORG50</td>
<td>Millipore</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>6-well plate</td>
<td>6 Well Cell Culture Cluster</td>
<td>Corning Incorporated</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Incubator</td>
<td>Forma Scientific CO2 Water Jacketed Incubator</td>
<td>Forma Scientific</td>
<td>&nbsp;</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>

preparation and culture of chicken auditory brainstem slices

Le tronc cérébral du poulet est un système modèle bien établi qui a été largement utilisée pour étudier l&#39;anatomie et la physiologie du système auditif à des périodes discrètes du développement de 1-4 ainsi que les mécanismes de codage temporel dans le système nerveux central 5-7. Nous présentons ici une méthode pour préparer les tranches de poulet auditifs du tronc cérébral qui peut être utilisé pour des procédures expérimentales aiguës ou de tranches de culture organotypique à long terme des manipulations expérimentales. Le tronc cérébral est composé de poulet angularis noyau, magnocellularis, olive laminaris et supérieures. Ces noyaux sont responsables du traitement des sons binauraux et unique préparation tranche coronale de préserver tout le circuit. Finalement, les cultures organotypique peut fournir l&#39;occasion de manipuler plusieurs paramètres du développement tels que l&#39;activité synaptique, l&#39;expression de composants pré-et post-synaptiques, l&#39;expression de l&#39;excitabilité des aspects de contrôle et de l&#39;expression différentielle des gènes Cette approche peut être utilisée pour élargir les connaissances générales sur le développement des circuits neuronaux, de raffinement et de la maturation.

Watch this Scientific Journal Video about Preparation and Culture of Chicken Auditory Brainstem Slices at JoVE.com

Preparation and Culture of Chicken Auditory Brainstem Slices

Le tronc cérébral du poulet est composé de noyaux responsable du traitement des sons binauraux. Une préparation seule tranche coronale maintient tout le circuit alors que l&#39;approche de culture fournit une préparation unique pour étudier le développement de la structure neuronale et de la fonction auditive au niveau moléculaire, cellulaire et réseau.

Préparation et de la culture de tranches de poulet auditive du tronc

The chicken auditory brainstem is a well-established model system that has been widely used to study the anatomy and physiology of auditory processing at ...

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