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

Aktuelle und ehemalige Rubel lab Mitglieder.

1. Vorbereitung der Dissecting Bereich <ol><li> Kontinuierlich Blase künstliche Rückenmarksflüssigkeit (ACSF) mit einer Mischung aus 95% O 2 / 5% CO 2 (pH-Wert zwischen 7,2 bis 7,4, Osmolarität 295-310 mOsm / l). </li><li> Während ACSF brodelt, saubere Arbeitsfläche mit 70% EtOH. Reinigen Sie auch die Vibratom und Schneidscheibe. Spülen Klinge mit destilliertem Wasser (dH 2 O). </li><li> Legen Sie saubere Flüssigkeitsaufnahme Pad auf Sezieren mit geeigneten Werkzeugen sezieren....

<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>

Seit mehreren Jahrzehnten hat sich die akuten Scheibe Vorbereitung des Huhns Hirnstamm verwendet worden, um Studie auditiven Verarbeitungs-9, 10. Dieser Ansatz hat eine enorme Menge von in-vitro physiologischen Daten auf binauralen Verarbeitung sowohl von Entwicklungs-und reifen Staaten 4, 11, 12 vorgesehen. Viel ist über diese hoch spezialisierten Schaltung und die Rolle jeder Kern spielt in der zeitlichen Verarbeitung von Ton 13, 14 bekannt. In der Tat haben kurzfristige expe...

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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

Das Huhn auditorischen Hirnstamm ist ein gut etabliertes Modell, das in großem Umfang verwendet wurde, um die Anatomie und Physiologie der auditiven Verarbeitung bei diskreten Perioden der Entwicklung 1-4 sowie Mechanismen für die zeitliche Kodierung in das zentrale Nervensystem 5-7 studieren. Hier präsentieren wir eine Methode, um Hühner auditorischen Hirnstamm Scheiben, die für die akute experimentelle Verfahren oder zur Kultur organotypischen Schnitten für die langfristige experimentelle Manipulationen genutzt werden kann vorzubereiten. Das Huhn auditorischen Hirnstamm ist der Kern angularis, magnocellularis, Laminaris und oberen Olive zusammen. Diese Kerne sind verantwortlich für die binaurale Signalverarbeitung und Single koronare Schicht Präparate erhalten die gesamte Schaltung. Letztlich kann organotypischen Kulturen die Möglichkeit bieten, mehrere Entwicklungs-Parameter wie synaptische Aktivität, Expression von Pre-und postsynaptischen Komponenten, Ausdruck von Aspekten Controlling Erregbarkeit und differenziellen Genexpression zu manipulieren Dieser Ansatz kann verwendet werden, um die allgemeinen Kenntnisse über neuronale Schaltkreis Entwicklung, Verfeinerung und Reifung zu erweitern.

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

Das Huhn auditorischen Hirnstamm ist der Kerne verantwortlich für die binaurale Signalverarbeitung zusammen. Ein einziger koronare Schicht Vorbereitung unterhält die gesamte Schaltung, während die kultivierten Ansatz bietet eine einzigartige Vorbereitung auf die Entwicklung der neuronalen Struktur und auditiven Funktion auf molekularer, zellulärer und Netzebenen zu studieren.

Vorbereitung und Kultur von Chicken auditorischen Hirnstamm Slices

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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Herstellung von oligomeren β-Amyloid 1-42 Und Induktion der synaptischen Plastizität Impairment auf Schnitten des Hippocampus

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