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

Attuali ed ex membri del laboratorio Rubel.

1. Preparazione di dissezione Area <ol><li> Continuamente bolla artificiale liquido cerebro-spinale (ACSF) con una miscela di 95% O 2 / 5% di CO 2 (pH compreso tra 7,2-7,4, osmolarità 295-310 mOsm / l). </li><li> Mentre ACSF è spumeggiante, pulire l&#39;area di lavoro con il 70% EtOH. Pulire anche il vibratome e lama di taglio. Risciacquare con acqua distillata lama (dH 2 O). </li><li> Luogo pulito pad assorbimento di liquidi sulla dissezione area con adeguati strumenti di dissezione...

<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>

Per diversi decenni, la preparazione fetta acuta del tronco cerebrale pollo è stato usato per studiare l&#39;elaborazione uditiva 9, 10. Questo approccio ha fornito una quantità enorme di dati in vitro fisiologici sul trattamento binaurale da entrambi gli stati di sviluppo e maturare 4, 11, 12. Si sa molto su questo circuito ad alta specializzazione e il ruolo di ogni nucleo svolge nella lavorazione temporale dei suoni 13, 14. In realtà, manipolazioni sperimentali a breve ter...

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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

Il tronco cerebrale uditivo pollo è un collaudato sistema di modello che è stato ampiamente usato per studiare l&#39;anatomia e la fisiologia del processo uditivo in periodi di discreto sviluppo 04/01 nonché meccanismi per la codifica temporale nel sistema nervoso centrale 5-7. Qui vi presentiamo un metodo per preparare le fette di pollo uditivi del tronco encefalico, che può essere utilizzato per acuti procedure sperimentali o di fette cultura organotipica per le manipolazioni sperimentali a lungo termine. Il tronco cerebrale uditivo pollo è composto da angularis nucleo, magnocellularis, oliva laminaris e superiore. Questi nuclei sono responsabili per l&#39;elaborazione del suono binaurale e singole preparazioni fetta coronale preservare l&#39;intero circuito. In definitiva, le culture fetta organotipica in grado di fornire l&#39;opportunità di manipolare vari parametri di sviluppo come l&#39;attività sinaptica, espressione di componenti pre e post-sinaptici, espressione di eccitabilità aspetti di controllo e di espressione genica differenziale Questo approccio può essere utilizzato per ampliare la conoscenza generale sullo sviluppo del circuito neurale, affinamento e maturazione.

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

Il tronco cerebrale uditivo pollo è costituita da nuclei responsabile per l&#39;elaborazione del suono binaurale. Una preparazione singola fetta coronale mantiene l&#39;intero circuito mentre l&#39;approccio colto fornisce una preparazione unica di studiare lo sviluppo della struttura neuronale e della funzione uditiva a livello molecolare, cellulare e di rete.

La preparazione e la cultura di fette di pollo uditive tronco

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