Name: tuning in the hippocampal theta band in vitro: methodologies for recording from the isolated rodent septohippocampal circuit
Uploaded: 2017-08-02T13:00:00
Description: Watch this Scientific Journal Video about Tuning in the Hippocampal Theta Band In Vitro: Methodologies for Recording from the Isolated Rodent Septohippocampal Circuit at JoVE.com

Questo lavoro è stato sostenuto dagli Istituti Canadesi di Salute e Scienze Naturali.

Tutte le procedure sono state eseguite secondo protocolli e linee guida approvate dal comitato McGill University Animal Care e dal Canadian Council on Animal Care. 1. Preparazione acuta di ippocampo in vitro NOTA: L&#39;isolamento della preparazione ippocampale intatta comporta tre fasi principali: (1) Preparazione di soluzioni e attrezzature, (2) Dissection dell&#39;ippocampo e (3) Impostazione del sistema di velocità di perfusione veloce necessaria per ...

<ol>
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E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Strategy+selection+in+a+task+with+spatial+and+nonspatial+components:+effects+of+fimbria-fornix+lesions+in+rats.">Strategy selection in a task with spatial and nonspatial components: effects of fimbria-fornix lesions in rats.</a> Behav Neural Biol. 58 (3), 171-179 (1992).</li><li>Osipova, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Theta+and+gamma+oscillations+predict+encoding+and+retrieval+of+declarative+memory.">Theta and gamma oscillations predict encoding and retrieval of declarative memory.</a> J Neurosci. 26 (28), 7523-7531 (2006).</li><li>Stumpf, C., Petsche, H., Gogolak, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+significance+of+the+rabbit's+septum+as+a+relay+station+between+the+midbrain+and+the+hippocampus.+II.+The+differential+influence+of+drugs+upon+both+the+septal+cell+firing+pattern+and+the+hippocampus+theta+activity.">The significance of the rabbit's septum as a relay station between the midbrain and the hippocampus. II. The differential influence of drugs upon both the septal cell firing pattern and the hippocampus theta activity.</a> Electroencephalogr Clin Neurophysiol. 14, 212-219 (1962).</li><li>Mitchell, S. J., Ranck, J. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Generation+of+theta+rhythm+in+medial+entorhinal+cortex+of+freely+moving+rats.">Generation of theta rhythm in medial entorhinal cortex of freely moving rats.</a> Brain Res. 189 (1), 49-66 (1980).</li><li>Alonso, A., Garcia-Austt, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuronal+sources+of+theta+rhythm+in+the+entorhinal+cortex+of+the+rat.+I.+Laminar+distribution+of+theta+field+potentials.">Neuronal sources of theta rhythm in the entorhinal cortex of the rat. I. Laminar distribution of theta field potentials.</a> Exp Brain Res. 67 (3), 493-501 (1987).</li><li>Vertes, R. P., Kocsis, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Brainstem-diencephalo-septohippocampal+systems+controlling+the+theta+rhythm+of+the+hippocampus.">Brainstem-diencephalo-septohippocampal systems controlling the theta rhythm of the hippocampus.</a> Neuroscience. 81 (4), 893-926 (1997).</li><li>Bland, B. H., Colom, L. V., Konopacki, J., Roth, S. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intracellular+records+of+carbachol-induced+theta+rhythm+in+hippocampal+slices.">Intracellular records of carbachol-induced theta rhythm in hippocampal slices.</a> Brain Res. 447 (2), 364-368 (1988).</li><li>Cobb, S. R., Buhl, E. H., Halasy, K., Paulsen, O., Somogyi, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synchronization+of+neuronal+activity+in+hippocampus+by+individual+GABAergic+interneurons.">Synchronization of neuronal activity in hippocampus by individual GABAergic interneurons.</a> Nature. 378 (6552), 75-78 (1995).</li><li>Williams, J. H., Kauer, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Properties+of+carbachol-induced+oscillatory+activity+in+rat+hippocampus.">Properties of carbachol-induced oscillatory activity in rat hippocampus.</a> J Neurophysiol. 78 (5), 2631-2640 (1997).</li><li>Chapman, C. A., Lacaille, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cholinergic+induction+of+theta-frequency+oscillations+in+hippocampal+inhibitory+interneurons+and+pacing+of+pyramidal+cell+firing.">Cholinergic induction of theta-frequency oscillations in hippocampal inhibitory interneurons and pacing of pyramidal cell firing.</a> J Neurosci. 19 (19), 8637-8645 (1999).</li><li>Strata, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intrinsic+oscillations+in+CA3+hippocampal+pyramids:+physiological+relevance+to+theta+rhythm+generation.">Intrinsic oscillations in CA3 hippocampal pyramids: physiological relevance to theta rhythm generation.</a> Hippocampus. 8 (6), 666-679 (1998).</li><li>Kocsis, B., Bragin, A., Buzsaki, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interdependence+of+multiple+theta+generators+in+the+hippocampus:+a+partial+coherence+analysis.">Interdependence of multiple theta generators in the hippocampus: a partial coherence analysis.</a> J Neurosci. 19 (14), 6200-6212 (1999).</li><li>Fellous, J. M., Sejnowski, T. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cholinergic+induction+of+oscillations+in+the+hippocampal+slice+in+the+slow+(0.5-2+Hz),+theta+(5-12+Hz),+and+gamma+(35-70+Hz)+bands.">Cholinergic induction of oscillations in the hippocampal slice in the slow (0.5-2 Hz), theta (5-12 Hz), and gamma (35-70 Hz) bands.</a> Hippocampus. 10 (2), 187-197 (2000).</li><li>Gillies, M. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+model+of+atropine-resistant+theta+oscillations+in+rat+hippocampal+area+CA1.">A model of atropine-resistant theta oscillations in rat hippocampal area CA1.</a> J Physiol. 543 (Pt 3), 779-793 (2002).</li><li>Gloveli, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthogonal+arrangement+of+rhythm-generating+microcircuits+in+the+hippocampus.">Orthogonal arrangement of rhythm-generating microcircuits in the hippocampus.</a> Proc Natl Acad Sci U S A. 102 (37), 13295-13300 (2005).</li><li>Konopacki, J., Eckersdorf, B., Kowalczyk, T., Golebiewski, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Firing+cell+repertoire+during+carbachol-induced+theta+rhythm+in+rat+hippocampal+formation+slices.">Firing cell repertoire during carbachol-induced theta rhythm in rat hippocampal formation slices.</a> Eur J Neurosci. 23 (7), 1811-1818 (2006).</li><li>Hajos, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Maintaining+network+activity+in+submerged+hippocampal+slices:+importance+of+oxygen+supply.">Maintaining network activity in submerged hippocampal slices: importance of oxygen supply.</a> Eur J Neurosci. 29 (2), 319-327 (2009).</li><li>Manseau, F., Goutagny, R., Danik, M., Williams, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+hippocamposeptal+pathway+generates+rhythmic+firing+of+GABAergic+neurons+in+the+medial+septum+and+diagonal+bands:+an+investigation+using+a+complete+septohippocampal+preparation+in+vitro.">The hippocamposeptal pathway generates rhythmic firing of GABAergic neurons in the medial septum and diagonal bands: an investigation using a complete septohippocampal preparation in vitro.</a> J Neurosci. 28 (15), 4096-4107 (2008).</li><li>Goutagny, R., Jackson, J., Williams, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Self-generated+theta+oscillations+in+the+hippocampus.">Self-generated theta oscillations in the hippocampus.</a> Nat Neurosci. 12 (12), 1491-1493 (2009).</li><li>Jackson, J., Goutagny, R., Williams, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fast+and+slow+gamma+rhythms+are+intrinsically+and+independently+generated+in+the+subiculum.">Fast and slow gamma rhythms are intrinsically and independently generated in the subiculum.</a> J Neurosci. 31 (34), 12104-12117 (2011).</li><li>Amilhon, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Parvalbumin+Interneurons+of+Hippocampus+Tune+Population+Activity+at+Theta+Frequency.">Parvalbumin Interneurons of Hippocampus Tune Population Activity at Theta Frequency.</a> Neuron. 86 (5), 1277-1289 (2015).</li><li>Huh, C. 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Mentre le registrazioni elettrofisiologiche da fette acute di ippocampo costituiscono una tecnica standard in vitro , i metodi presentati qui differiscono sostanzialmente dal metodo classico. A differenza delle preparazioni sottili di fetta dove sono visibili strati cellulari specifici sulla superficie e possono essere esaminati direttamente, i preparati ippocampali intatti sono più simili alle configurazioni in vivo in cui gli elettrodi vengono abbassati in regioni cerebrali mirate attraversando sing...

Le oscillazioni di theta ippocampali (4-12 Hz) sono tra le forme più prevalenti di attività ritmica nel cervello dei mammiferi e si credono a svolgere ruoli chiave nelle funzioni cognitive quali elaborazione di informazioni spaziali e formazione di memorie episodiche 1 , 2 , 3 . Mentre diversi studi in vivo che evidenziano il rapporto delle cellule place-theta modulate con studi di navigazione spaziale e lesioni, nonché prove cliniche, supportano la visione che le oscillazioni ippocampali sono coinvolte nella formazione della memoria 4 , 5 , 6 , i meccanismi associati Con generazione di oscillazioni theta ippocampali non sono ancora completamente compresi. Le prime indagini in vivo suggerivano che l&#39;attività theta dipendesse principalmente da oscillatori estrinseci, in particolare l&#39;ingresso ritmicoDa strutture cerebrali afferenti come il settto e la corteccia entorhina 7 , 8 , 9 , 10 . È stato anche postulato un ruolo per fattori intrinseci - connettività interna delle reti neuronali ippocampali insieme alle proprietà dei neuroni ippocampali - in base alle osservazioni in vitro 11 , 12 , 13 , 14 , 15 , 16 , 17 , 18 . Tuttavia, a parte alcuni studi di rilievo 19 , 20 , 21 , difficoltà nello sviluppo di approcci che potrebbero replicare attività fisiologicamente realistiche di popolazione in una semplice preparazione di fetta in vitroSono stati, per lungo tempo, ritardato un esame sperimentale più dettagliato delle abilità intrinseche dell&#39;ippocampo e delle aree correlate per auto generare oscillazioni di theta. Un importante inconveniente dell&#39;insieme sperimentale in vitro di sottile fetta è che l&#39;organizzazione cellulare e sinaptica 3D delle strutture cerebrali è di solito compromessa. Ciò significa che non possono essere supportate molte forme di attività di rete concertata basate su gruppi di cellule distribuite spazialmente, che vanno da gruppi localizzati (≤1 mm radius) alle popolazioni di neuroni diffusi in una o più aree del cervello (&gt; 1 mm). Tenuto conto di queste considerazioni, è stato necessario un approccio diverso per studiare come le oscillazioni di theta emergono nell&#39;ippocampo e si propagino alle strutture di uscita corticali e subcorticali correlate. Negli ultimi anni, lo sviluppo iniziale della preparazione completa &quot;septo-ippocampale&quot; per esaminare l&#39;intero bidirezionaleLe due delle strutture 22 e la conseguente evoluzione della preparazione &quot;ippocampo isolato&quot; hanno rivelato che le oscillazioni di theta intrinseca si verificano spontaneamente nell&#39;ippocampo privo di input ritmico esterno 23 . Il valore di questi approcci risiede sulla prima comprensione che tutta la struttura funzionale di queste regioni doveva essere conservata per poter funzionare come un ritmo di theta in vitro 22 .

<table border="0" cellpadding="0" cellspacing="0" width="820">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:319px;">Reagents</td>
			<td style="width:135px;"></td>
			<td style="width:119px;"></td>
			<td style="width:247px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sodium Chloride</td>
			<td>Sigma Aldrich</td>
			<td>S9625</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sucrose</td>
			<td>Sigma Aldrich</td>
			<td>S9378</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sodium Bicarbonate</td>
			<td>Sigma Aldrich</td>
			<td>S5761</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">NaH2PO4 - sodium phosphate monobasic</td>
			<td>Sigma Aldrich</td>
			<td>S8282</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Magnesium sulfate</td>
			<td>Sigma Aldrich</td>
			<td>M7506</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Potassium Chloride</td>
			<td>Sigma Aldrich</td>
			<td>P3911</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">D-(+)-Glucose</td>
			<td>Sigma Aldrich</td>
			<td>G7528</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Calcium chloride dihydrate</td>
			<td>Sigma Aldrich</td>
			<td>C5080</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sodium Ascorbate</td>
			<td>Sigma Aldrich</td>
			<td>A7631-25G</td>
			<td></td>
		</tr>
		<tr height="30">
			<td height="30" style="height:30px;width:319px;">Name</td>
			<td style="width:135px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Equipment</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Standard Dissecting Scissors</td>
			<td>Fisher Scientific</td>
			<td>08-951-25</td>
			<td>brain extraction</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Scalpel Handle #4, 14cm</td>
			<td>WPI</td>
			<td>500237</td>
			<td>brain extraction</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Filter forceps, flat jaws, straight (11cm)</td>
			<td>WPI</td>
			<td>500456</td>
			<td>brain extraction</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Paragon Stainless Steel Scalpel Blades #20</td>
			<td>Ultident</td>
			<td>02-90010-20</td>
			<td>brain extraction</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fine Point Curved Dissecting Scissors</td>
			<td>Thermo Fisher Scientific</td>
			<td>711999</td>
			<td>brain extraction</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Teflon (PTFE) -coated thin spatula</td>
			<td>VWR</td>
			<td>82027-534</td>
			<td>hippocampal preparation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hayman Style Microspatula</td>
			<td>Fisher Scientific</td>
			<td>21-401-25A</td>
			<td>hippocampal preparation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Lab spoon</td>
			<td>Fisher Scientific</td>
			<td>14-375-20</td>
			<td>hippocampal preparation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Borosilicate Glass Pasteur Pipets</td>
			<td>Fisher Scientific</td>
			<td>13-678-20A</td>
			<td>hippocampal preparation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Droper</td>
			<td>Fisher Scientific</td>
			<td></td>
			<td>hippocampal preparation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Razor blades Single edged</td>
			<td>VWR</td>
			<td>55411-055</td>
			<td>hippocampal preparation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Lens paper (4X6 inch)</td>
			<td>VWR</td>
			<td>52846-001</td>
			<td>hippocampal preparation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Glass petri dishes (100 x 20 mm)</td>
			<td>VWR</td>
			<td>25354-080</td>
			<td>hippocampal preparation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Plastic tray for ice; size 30 x 20 x 5 cm</td>
			<td>n.a.</td>
			<td>n.a.</td>
			<td>hippocampal preparation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Single Inline Solution Heater</td>
			<td>Warner Instruments</td>
			<td>SH-27B</td>
			<td>perfusion system</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Aquarium air stones for bubbling</td>
			<td>n.a.</td>
			<td>n.a.</td>
			<td>perfusion system</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Tygon E-3603 tubing (ID 1/16 OD 1/8)</td>
			<td>Fisherbrand</td>
			<td>14-171-129</td>
			<td>perfusion system</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Electric Skillet</td>
			<td>Black &#38; Decker</td>
			<td>n.a.</td>
			<td>perfusion system</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">95% O2/5% CO2 gas mixture (carbogen)&#160;</td>
			<td>Vitalaire</td>
			<td>SG466204A</td>
			<td>perfusion system</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Glass bottles/flasks (4 x 1 L)</td>
			<td>n.a.</td>
			<td>n.a.</td>
			<td>perfusion system</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Submerged recording Chamber</td>
			<td>custom design (FM)</td>
			<td>n.a.</td>
			<td style="width:247px;">Commercial alternative may be used</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Glass pipettes (1.5 / 0.84 OD/ID (mm) )</td>
			<td>WPI</td>
			<td>1B150F-4</td>
			<td>electrophysiology</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hum Bug 50/60 Hz Noise Eliminator</td>
			<td>Quest Scientific</td>
			<td>Q-Humbug</td>
			<td>electrophysiology</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Multiclamp 700B patch-clamp amplifier</td>
			<td>Molecular devices</td>
			<td>MULTICLAMP</td>
			<td>electrophysiology</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Multiclamp 700B Commander Program</td>
			<td>Molecular devices</td>
			<td>MULTICLAMP</td>
			<td>electrophysiology</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Digital/Analogue converter</td>
			<td>Molecular devices</td>
			<td>DDI440</td>
			<td>electrophysiology</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PCLAMP10</td>
			<td>Molecular devices</td>
			<td>PCLAMP10</td>
			<td>electrophysiology</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Vibration isolation table&#160;</td>
			<td>Newport</td>
			<td>n.a.</td>
			<td>electrophysiology</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Micromanipulators (manually operated )</td>
			<td>Siskiyou&#160;</td>
			<td>MX130</td>
			<td>electrophysiology (LFP)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Micromanipulators (automated)</td>
			<td>Siskiyou&#160;</td>
			<td>MC1000e</td>
			<td>electrophysiology (patch)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Audio monitor&#160;</td>
			<td>A-M Systems</td>
			<td>Model 3300</td>
			<td>electrophysiology</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Micropipette/Patch pipette puller</td>
			<td>Sutter</td>
			<td>P-97</td>
			<td>electrophysiology</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Custom-built upright fluorescence microscope</td>
			<td>Siskiyou</td>
			<td>n.a.</td>
			<td>Imaging</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Analogue video camera</td>
			<td>COHU</td>
			<td>4912-2000/0000</td>
			<td>Imaging</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Digital frame grabber with imaging software</td>
			<td>EPIX, Inc</td>
			<td>PIXCI-SV7</td>
			<td>Imaging</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Olympus 2.5x objective</td>
			<td>Olympus</td>
			<td>MPLFLN</td>
			<td>Imaging</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Olympus 40x water immersion objective</td>
			<td>Olympus</td>
			<td>UIS2 LUMPLFLN</td>
			<td>Imaging</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Custom-made light-emitting diode (LED) system&#160;</td>
			<td>custom</td>
			<td>n.a.</td>
			<td>optogenetic stimulation (Amhilon et al., 2015)</td>
		</tr>
		<tr height="30">
			<td height="30" style="height:30px;width:319px;">Name</td>
			<td style="width:135px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Animals</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:319px;">PV::Cre (KI) mice</td>
			<td style="width:135px;">Jackson Laboratory</td>
			<td style="width:119px;">stock number 008069</td>
			<td style="width:247px;">Allow&#160; Cre-directed gene expression in PV interneurons</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:319px;">Constitutive-conditional Ai9 mice (R26-lox-stop-lox-tdTomato (KI))</td>
			<td style="width:135px;">Jackson Laboratory</td>
			<td style="width:119px;">stock number 007905</td>
			<td style="width:247px;">Express TdTomato following Cre-mediated recombination</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:319px;">Ai32 mice (R26-lox-stop-lox-ChR2(H134R)-EYFP</td>
			<td style="width:135px;">Jackson Laboratory</td>
			<td style="width:119px;">stock 
			number 012569</td>
			<td style="width:247px;">Express the improved channelrhodopsin-2/EYFP fusion protein following exposure to Cre recombinase</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PVChY mice</td>
			<td>In house breeding</td>
			<td>n.a.</td>
			<td style="width:247px;">Offspring obtained from cross-breeding the PV-Cre line with Ai32 mice (R26-lox-stop-lox-ChR2(H134R)-EYFP</td>
		</tr>
	</tbody>
</table>

tuning in the hippocampal theta band in vitro: methodologies for recording from the isolated rodent septohippocampal circuit

Questo protocollo descrive le procedure per la preparazione e la registrazione dall&#39;ippocampo intero isolato, dai topi transgenici e dai topi transgenici, nonché dai recenti miglioramenti nelle metodologie e nelle applicazioni per lo studio delle oscillazioni. Viene presentata una semplice caratterizzazione della preparazione ippocampale isolata, in cui viene analizzata la relazione tra oscillatori ippocampali theta e l&#39;attività delle cellule piramidali e degli interneuroni GABAergici delle aree cornu ammonis-1 (CA1) e subiculum (SUB). Nel complesso mostriamo che l&#39;ippocampo isolato è in grado di generare oscillazioni di theta intrinseche in vitro e che la ritmicità generata all&#39;interno dell&#39;ippocampo può essere manipolata in modo preciso dalla stimolazione ottogenetica di interneuroni parvalbuminici (PV). Il preparato ippocampo isolato in vitro offre un&#39;occasione unica per utilizzare registrazioni simultanee di campo e intracellulari da un patch-clamp da identificazioni visivamente identificatePer capire meglio i meccanismi che sottendono alla generazione del ritmo theta.

<html> Questa sezione illustra esempi di risultati che possono essere ottenuti studiando le oscillazioni di theta nella preparazione ippocampale isolata in mouse in vitro . La procedura di dissezione per estrarre l&#39;ippocampo isolato è illustrata in Figura 1 . Utilizzando questa preparazione, le oscillazioni di theta intrinseca possono essere esaminate durante il posizionamento di elettrodi di campo multiplo, registrando l&#39;attività complessi...

Watch this Scientific Journal Video about Tuning in the Hippocampal Theta Band In Vitro: Methodologies for Recording from the Isolated Rodent Septohippocampal Circuit at JoVE.com

Tuning in the Hippocampal Theta Band In Vitro: Methodologies for Recording from the Isolated Rodent Septohippocampal Circuit

Qui presentiamo un protocollo per registrare la teoria ritmica della rete neuronale e le oscillazioni gamma da una preparazione interamente isolata di ippocampo. Descriviamo le fasi sperimentali dall&#39;estrazione dell&#39;ippocampo ai dettagli del campo, delle registrazioni a blocchi di patch patch unitari e intere cellule, nonché della stimolazione ottogenetica del ritmo theta.

Tuning nella Banda Theta Ippocampale<em&gt; In vitro</em&gt;: Metodologie per la registrazione dal circuito isolante di roditore Septohippocampal

This protocol outlines the procedures for preparing and recording from the isolated whole hippocampus, of WT and transgenic mice, along with recent ...

Research

JoVE Journal

Neuroscience

Tuning in the Hippocampal Theta Band In Vitro: Methodologies for Recording from the Isolated Rodent Septohippocampal Circuit

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

Retrograda Etichettatura fluorescente Consente mirata extracellulare singola unità di registrazione da parte dei neuroni identificati<em&gt; In vivo</em

The overall goal of this method is to record single-unit responses from an identified population of neurons. In vivo electrophysiological recordings from ...

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Investigations on Alterations of Hippocampal Circuit Function Following Mild Traumatic Brain Injury

Indagini su Alterazioni della funzione del circuito ippocampale seguito lieve trauma cranico

Traumatic Brain Injury (TBI) afflicts more than 1.7 million people in the United States each year and even mild TBI can lead to persistent neurological ...

Dual Electrophysiological Recordings of Synaptically-evoked Astroglial and Neuronal Responses in Acute Hippocampal Slices

Due registrazioni elettrofisiologiche di sinapticamente-evocati Risposte astrogliale e neuronale in acute fettine di ippocampo

Astrocytes form together with neurons tripartite synapses, where they integrate and modulate neuronal activity. Indeed, astrocytes sense neuronal inputs ...

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Multi-unit Recording Methods to Characterize Neural Activity in the Locust Schistocerca Americana Olfactory Circuits

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Detection and interpretation of olfactory cues are critical for the survival  of many organisms. Remarkably, species across phyla have strikingly similar ...

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Consegna diretta intraventricolare di farmaci per il sistema nervoso centrale, Roditore

Due to an inability to cross the blood brain barrier, certain drugs need to be directly delivered into the central nervous system (CNS). Our lab focuses ...

An In-vitro Preparation of Isolated Enteric Neurons and Glia from the Myenteric Plexus of the Adult Mouse

An In-vitro Preparation of Isolated Enteric Neurons and Glia from the Myenteric Plexus of the Adult Mouse

Un<em&gt; In-vitro</em&gt; Preparazione dei neuroni enterici isolati e glia dal plesso mienterico del topo adulto

The enteric nervous system is a vast network of neurons and glia running the length of the gastrointestinal tract that functionally controls ...

Simultaneous Electrophysiological Recording and Micro-injections of Inhibitory Agents in the Rodent Brain

Registrazione simultanea elettrofisiologici e micro-iniezioni di agenti inibitori nel cervello del roditore

Here we describe a method for the construction of a single-use &#8220;injectrode&#8221; using commercially accessible and affordable parts. A probing ...

Utilizing Combined Methodologies to Define the Role of Plasma Membrane Delivery During Axon Branching and Neuronal Morphogenesis

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During neural development, growing axons extend to multiple synaptic partners by elaborating axonal branches. Axon branching is promoted by extracellular ...

Recording Gamma Band Oscillations in Pedunculopontine Nucleus Neurons

Registrazione Gamma banda oscillazioni Pedunculopontine Nucleo neuroni

Synaptic efferents from the PPN are known to modulate the neuronal activity of several intralaminar thalamic regions (e.g., the ...

Recording Synaptic Plasticity in Acute Hippocampal Slices Maintained in a Small-volume Recycling-, Perfusion-, and Submersion-type Chamber System

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Even though experiments on brain slices have been in use since 1951, problems remain that reduce the probability of achieving a stable and successful ...