この研究は、カナダの健康研究および自然科学研究所によって支援されました。

<ol>
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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. Y., 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=Excitatory+Inputs+Determine+Phase-Locking+Strength+and+Spike-Timing+of+CA1+Stratum+Oriens/Alveus+Parvalbumin+and+Somatostatin+Interneurons+during+Intrinsically+Generated+Hippocampal+Theta+Rhythm.">Excitatory Inputs Determine Phase-Locking Strength and Spike-Timing of CA1 Stratum Oriens/Alveus Parvalbumin and Somatostatin Interneurons during Intrinsically Generated Hippocampal Theta Rhythm.</a> J Neurosci. 36 (25), 6605-6622 (2016).</li><li>Gu, 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=NMDA-dependent+phase+synchronization+between+septal+and+temporal+CA3+hippocampal+networks.">NMDA-dependent phase synchronization between septal and temporal CA3 hippocampal networks.</a> J Neurosci. 33 (19), 8276-8287 (2013).</li><li>Jackson, 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=Reversal+of+theta+rhythm+flow+through+intact+hippocampal+circuits.">Reversal of theta rhythm flow through intact hippocampal circuits.</a> Nat Neurosci. 17 (10), 1362-1370 (2014).</li><li>Gonzalez-Burgos, G., Lewis, D. 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著者は競合する商業的または財政的利益を宣言していない。

急性海馬スライスからの電気生理学的記録は標準的なインビトロ技術を構成するが、ここに提示される方法は従来のアプローチとは実質的に異なる。特定の細胞層が表面で可視であり、直接検査することができる薄いスライス調製物とは異なり、インタクトな海馬調製物は、個々の層を横切って電極を標的脳領域に下ろすインビボ配置にさらに類似している。海馬の完全?...

海馬のシータ振動（4から12 Hz）は哺乳動物の脳におけるリズム活性の最も優勢な形態の中で、そのようなエピソードメモリ1、2、3の時空間情報及び形成の処理として、認知機能に重要な役割を果たしていると考えられています。空間ナビゲーションと病変の研究、ならびに臨床的証拠を有するシータ変調場所細胞の関係を強調いくつかのin vivo研究は、海馬のシータ振動が記憶形成4、5、6、関連するメカニズムに関与しているという見解を支持しながら海馬の生成は、まだ完全に理解されていない。初期のインビボでの研究では、シータ活性は主として外因性オシレータ、特にリズム入力に依存していることが示唆された求心性脳構造から、このような隔壁及び嗅内皮質7、8、9、10として。一緒に海馬ニューロンの特性を有する海馬ニューラルネットワークの内部接続- -内因性因子の役割はまた、in vitroの観察11、12、13、14、15、16、17、18に基づいて仮定されました。しかし、離れていくつかの画期的な研究19、20、21、 体外スライス標本でシンプルに生理的に現実的な人口の活動を複製することができなアプローチを開発する上での困難から長い間、海馬の固有の能力および関連領域のより詳細な実験的検討が、シータ振動を自己生成するのを遅らせてしまった。 インビトロ薄切片実験の標準的な設定の重要な欠点は、脳構造の3D細胞およびシナプス組織が通常損なわれていることである。これは、局所的なグループ（半径1mm以下）から1つ以上の脳領域（&gt; 1mm）にわたって広がるニューロンの集団に及ぶ、空間的に分布した細胞アセンブリに基づく協調的ネットワーク活動の多くの形態がサポートされないことを意味する。これらの考察を考慮して、シータ振動が海馬でどのように現れ、関連する皮質および皮質下の出力構造に伝播するかを研究するために、異なるタイプのアプローチが必要であった。 近年、双方向性を調べるための「完全なsepto-hippocampal」準備の初期の開発2つの構造22の構造、およびその後の「単離された海馬」調製の進化は、固有のシータ振動が外的な律動的な入力23を欠く海馬において自発的に起こることを明らかにした。これらのアプローチの値は、これらの領域の全体の機能構成は、インビトロ 22 におけるシータリズム発生器として機能するために保存しなければならなかった最初の洞察です。

<table><tbody><tr><td>&lt;strong&gt;Reagents&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Sodium Chloride</td><td>Sigma Aldrich</td><td>S9625</td><td></td></tr><tr><td>Sucrose</td><td>Sigma Aldrich</td><td>S9378</td><td></td></tr><tr><td>Sodium Bicarbonate</td><td>Sigma Aldrich</td><td>S5761</td><td></td></tr><tr><td>NaH&lt;sub&gt;2&lt;/sub&gt;PO&lt;sub&gt;4&lt;/sub&gt; - sodium phosphate monobasic</td><td>Sigma Aldrich</td><td>S8282</td><td></td></tr><tr><td>Magnesium sulfate</td><td>Sigma Aldrich</td><td>M7506</td><td></td></tr><tr><td>Potassium Chloride</td><td>Sigma Aldrich</td><td>P3911</td><td></td></tr><tr><td>D-(+)-Glucose</td><td>Sigma Aldrich</td><td>G7528</td><td></td></tr><tr><td>Calcium chloride dihydrate</td><td>Sigma Aldrich</td><td>C5080</td><td></td></tr><tr><td>Sodium Ascorbate</td><td>Sigma Aldrich</td><td>A7631-25G</td><td></td></tr><tr><td>&lt;strong&gt;Name&lt;/strong&gt;</td><td>&lt;strong&gt;Company&lt;/strong&gt;</td><td>&lt;strong&gt;Catalog Number&lt;/strong&gt;</td><td>&lt;strong&gt;Comments&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;Equipment&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Standard Dissecting Scissors</td><td>Fisher Scientific</td><td>08-951-25</td><td>brain extraction</td></tr><tr><td>Scalpel Handle #4, 14 cm</td><td>WPI</td><td>500237</td><td>brain extraction</td></tr><tr><td>Filter forceps, flat jaws, straight (11 cm)</td><td>WPI</td><td>500456</td><td>brain extraction</td></tr><tr><td>Paragon Stainless Steel Scalpel Blades #20</td><td>Ultident</td><td>02-90010-20</td><td>brain extraction</td></tr><tr><td>Fine Point Curved Dissecting Scissors</td><td>Thermo Fisher Scientific</td><td>711999</td><td>brain extraction</td></tr><tr><td>Teflon (PTFE) -coated thin spatula</td><td>VWR</td><td>82027-534</td><td>hippocampal preparation</td></tr><tr><td>Hayman Style Microspatula</td><td>Fisher Scientific</td><td>21-401-25A</td><td>hippocampal preparation</td></tr><tr><td>Lab spoon</td><td>Fisher Scientific</td><td>14-375-20</td><td>hippocampal preparation</td></tr><tr><td>Borosilicate Glass Pasteur Pipets</td><td>Fisher Scientific</td><td>13-678-20A</td><td>hippocampal preparation</td></tr><tr><td>Droper</td><td>Fisher Scientific</td><td></td><td>hippocampal preparation</td></tr><tr><td>Razor blades Single edged</td><td>VWR</td><td>55411-055</td><td>hippocampal preparation</td></tr><tr><td>Lens paper (4 x 6&quot;)</td><td>VWR</td><td>52846-001</td><td>hippocampal preparation</td></tr><tr><td>Glass petri dishes (100 x 20 mm)</td><td>VWR</td><td>25354-080</td><td>hippocampal preparation</td></tr><tr><td>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><td>Single Inline Solution Heater</td><td>Warner Instruments</td><td>SH-27B</td><td>perfusion system</td></tr><tr><td>Aquarium air stones for bubbling</td><td>n.a.</td><td>n.a.</td><td>perfusion system</td></tr><tr><td>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><td>Electric Skillet</td><td>Black &amp;amp; Decker</td><td>n.a.</td><td>perfusion system</td></tr><tr><td>95% O&lt;sub&gt;2&lt;/sub&gt;/5% CO&lt;sub&gt;2&lt;/sub&gt; gas mixture (carbogen)&amp;nbsp;</td><td>Vitalaire</td><td>SG466204A</td><td>perfusion system</td></tr><tr><td>Glass bottles/flasks (4 x 1 L)</td><td>n.a.</td><td>n.a.</td><td>perfusion system</td></tr><tr><td>Submerged recording Chamber</td><td>custom design (FM)</td><td>n.a.</td><td>Commercial alternative may be used</td></tr><tr><td>Glass pipettes (1.5/0.84 OD/ID (mm) )</td><td>WPI</td><td>1B150F-4</td><td>electrophysiology</td></tr><tr><td>Hum Bug 50/60 Hz Noise Eliminator</td><td>Quest Scientific</td><td>Q-Humbug</td><td>electrophysiology</td></tr><tr><td>Multiclamp 700B patch-clamp amplifier</td><td>Molecular devices</td><td>MULTICLAMP</td><td>electrophysiology</td></tr><tr><td>Multiclamp 700B Commander Program</td><td>Molecular devices</td><td>MULTICLAMP</td><td>electrophysiology</td></tr><tr><td>Digital/Analogue converter</td><td>Molecular devices</td><td>DDI440</td><td>electrophysiology</td></tr><tr><td>PCLAMP10</td><td>Molecular devices</td><td>PCLAMP10</td><td>electrophysiology</td></tr><tr><td>Vibration isolation table&amp;nbsp;</td><td>Newport</td><td>n.a.</td><td>electrophysiology</td></tr><tr><td>Micromanipulators (manually operated )</td><td>Siskiyou&amp;nbsp;</td><td>MX130</td><td>electrophysiology (LFP)</td></tr><tr><td>Micromanipulators (automated)</td><td>Siskiyou&amp;nbsp;</td><td>MC1000e</td><td>electrophysiology (patch)</td></tr><tr><td>Audio monitor&amp;nbsp;</td><td>A-M Systems</td><td>Model 3300</td><td>electrophysiology</td></tr><tr><td>Micropipette/Patch pipette puller</td><td>Sutter</td><td>P-97</td><td>electrophysiology</td></tr><tr><td>Custom-built upright fluorescence microscope</td><td>Siskiyou</td><td>n.a.</td><td>Imaging</td></tr><tr><td>Analogue video camera</td><td>COHU</td><td>4912-2000/0000</td><td>Imaging</td></tr><tr><td>Digital frame grabber with imaging software</td><td>EPIX, Inc</td><td>PIXCI-SV7</td><td>Imaging</td></tr><tr><td>Olympus 2.5X objective</td><td>Olympus</td><td>MPLFLN</td><td>Imaging</td></tr><tr><td>Olympus 40X water immersion objective</td><td>Olympus</td><td>UIS2 LUMPLFLN</td><td>Imaging</td></tr><tr><td>Custom-made Light-Emitting Diode (LED) system&amp;nbsp;</td><td>custom</td><td>n.a.</td><td>optogenetic stimulation (Amhilon et al., 2015)</td></tr><tr><td>&lt;strong&gt;Name&lt;/strong&gt;</td><td>&lt;strong&gt;Company&lt;/strong&gt;</td><td>&lt;strong&gt;Catalog Number&lt;/strong&gt;</td><td>&lt;strong&gt;Comments&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;Animals&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>PV::Cre (KI) mice</td><td>Jackson Laboratory</td><td>stock number 008069</td><td>Allow&amp;nbsp; Cre-directed gene expression in PV interneurons</td></tr><tr><td>Constitutive-conditional Ai9 mice (R26-lox-stop-lox-tdTomato (KI))</td><td>Jackson Laboratory</td><td>stock number 007905</td><td>Express TdTomato following Cre-mediated recombination</td></tr><tr><td>Ai32 mice (R26-lox-stop-lox-ChR2(H134R)-EYFP</td><td>Jackson Laboratory</td><td>stock&lt;br/&gt; number 012569</td><td>Express the improved channelrhodopsin-2/EYFP fusion protein following exposure to Cre recombinase</td></tr><tr><td>PVChY mice</td><td>In house breeding</td><td>n.a.</td><td>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

このプロトコールは、WTおよびトランスジェニックマウスの単離された全海馬からの調製および記録のための手順、ならびにシータ振動の研究のための最近の方法論および適用の概要を概説する。単離された海馬調製物の簡単な特徴付けが提示され、それにより、角膜アンモニア-1（CA1）および漿膜（SUB）領域のピラミッド細胞およびGABA作動性介在ニューロンの活性とともに、内部海馬θオシレータ間の関係が調べられる。全体的に、本発明者らは、単離された海馬がインビトロで固有のシータ振動を生成することができ、海馬内で生成されたリズムが、パルブアルブミン陽性（PV）介在ニューロンの光発生刺激によって正確に操作され得ることを示す。 インビトロで単離された海馬調製物は、視覚的に同定されたneuからの同時のフィールドおよび細胞内パッチクランプ記録を使用する独特の機会を提供するシータ・リズムの生成メカニズムの理解を深めています。

<html>このセクションは、インビトロでマウス単離された海馬調製物におけるシータ振動を研究することによって得られ得る結果の例を説明する。単離された海馬を抽出するための解剖手順を図1に示す 。この準備を使用して、複数のフィールド電極の配置中に固有のシータ振動を検査し、全体的な活動を記録?...

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

ここでは、分離された全海馬調製物からのリズミカルなニューロンネットワークのシータおよびガンマ振動を記録するためのプロトコルを提示する。我々は、海馬の抽出からフィールド、ユニタリーおよび全細胞のパッチクランプ記録ならびにテータリズムの光発生ペーシングの詳細までの実験ステップを記載する。

海馬シータバンドのチューニング<em&gt;インビトロ</em&gt;：隔離されたげっ歯類胸腺腫回路からの記録方法

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

tuning-hippocampal-theta-band-vitro-methodologies-for-recording-from

Research

JoVE Journal

Neuroscience

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

9.7K ビュー.  McGill University. ここでは、分離された全海馬調製物からのリズミカルなニューロンネットワークのシータおよびガンマ振動を記録するためのプロトコルを提示する。我々は、海馬の抽出からフィールド、ユニタリーおよび全細胞のパッチクランプ記録ならびにテータリズムの光発生ペーシングの詳細までの実験ステップを記載する。 

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

Automatic Detection of Highly Organized Theta Oscillations in the Murine EEG

Theta activity is generated in the septohippocampal system and can be recorded using deep intrahippocampal electrodes and implantable electroencephalography (EEG) radiotelemetry or tether system approaches. Pharmacologically, hippocampal theta is heterogeneous (see dualistic theory) and can be differentiated into type I and type II theta. These individual EEG subtypes are related to specific cognitive and behavioral states, such as arousal, exploration, learning and memory, higher integrative functions, etc. In neurodegenerative diseases such as Alzheimer&#39;s, structural and functional alterations of the septohippocampal system can result in impaired theta activity/oscillations. A standard quantitative analysis of the hippocampal EEG includes a Fast-Fourier-Transformation (FFT)-based frequency analysis. However, this procedure does not provide details about theta activity in general and highly-organized theta oscillations in particular. In order to obtain detailed information on highly-organized theta oscillations in the hippocampus, we have developed a new analytical approach. This approach allows for time- and cost-effective quantification of the duration of highly-organized theta oscillations and their frequency characteristics.

Theta activity in the hippocampus is related to specific cognitive and behavioral stages. Here, we describe an analytical method to detect highly-organized theta oscillations within the hippocampus using a time-frequency (i.e., wavelet analysis)-based approach.

マウス脳波で高度に組織化されたシータ振動の自動検出

Theta activity is generated in the septohippocampal system and can be recorded using deep intrahippocampal electrodes and implantable ...

行動学

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

Astrocytes form together with neurons tripartite synapses, where they integrate and modulate neuronal activity. Indeed, astrocytes sense neuronal inputs through activation of their ion channels and neurotransmitter receptors, and process information in part through activity-dependent release of gliotransmitters. Furthermore, astrocytes constitute the main uptake system for glutamate, contribute to potassium spatial buffering, as well as to GABA clearance. These cells therefore constantly monitor synaptic activity, and are thereby sensitive indicators for alterations in synaptically-released glutamate, GABA and extracellular potassium levels. Additionally, alterations in astroglial uptake activity or buffering capacity can have severe effects on neuronal functions, and might be overlooked when characterizing physiopathological situations or knockout mice. Dual recording of neuronal and astroglial activities is therefore an important method to study alterations in synaptic strength associated to concomitant changes in astroglial uptake and buffering capacities. Here we describe how to prepare hippocampal slices, how to identify stratum radiatum astrocytes, and how to record simultaneously neuronal and astroglial electrophysiological responses. Furthermore, we describe how to isolate pharmacologically the synaptically-evoked astroglial currents.

The preparation of acute brain slices from isolated hippocampi, as well as the simultaneous electrophysiological recordings of astrocytes and neurons in stratum radiatum during stimulation of schaffer collaterals is described. The pharmacological isolation of astroglial potassium and glutamate transporter currents is demonstrated.

急性海馬スライスにおけるシナプス誘発アストログリアとニューロン活動の二重の電気生理学的記録

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

神経科学

Improved Preparation and Preservation of Hippocampal Mouse Slices for a Very Stable and Reproducible Recording of Long-term Potentiation

Long-term potentiation (LTP) is a type of synaptic plasticity characterized by an increase in synaptic strength and believed to be involved in memory encoding. LTP elicited in the CA1 region of acute hippocampal slices has been extensively studied. However the molecular mechanisms underlying the maintenance phase of this phenomenon are still poorly understood. This could be partly due to the various experimental conditions used by different laboratories. Indeed, the maintenance phase of LTP is strongly dependent on external parameters like oxygenation, temperature and humidity. It is also dependent on internal parameters like orientation of the slicing plane and slice viability after dissection.
The optimization of all these parameters enables the induction of a very reproducible and very stable long-term potentiation. This methodology offers the possibility to further explore the molecular mechanisms involved in the stable increase in synaptic strength in hippocampal slices. It also highlights the importance of experimental conditions in in vitro investigation of neurophysiological phenomena.

This paper presents a complete methodology to prepare and preserve in vitro acute hippocampal slices from adult mice. This protocol allows the recording of very stable long-lasting long-term potentiation (LTP) for more than 8 hours with a success rate of 95%.

長期増強の非常に安定しており、再現性録音のための改良された準備と海馬マウススライスの保全

Long-term potentiation (LTP) is a type of synaptic plasticity characterized by an increase in synaptic strength and believed to be involved in memory ...

Whole-cell Patch-clamp Recordings from Morphologically- and Neurochemically-identified Hippocampal Interneurons

GABAergic inhibitory interneurons play a central role within neuronal circuits of the brain. Interneurons comprise a small subset of the neuronal population (10-20%), but show a high level of physiological, morphological, and neurochemical heterogeneity, reflecting their diverse functions. Therefore, investigation of interneurons provides important insights into the organization principles and function of neuronal circuits. This, however, requires an integrated physiological and neuroanatomical approach for the selection and identification of individual interneuron types. Whole-cell patch-clamp recording from acute brain slices of transgenic animals, expressing fluorescent proteins under the promoters of interneuron-specific markers, provides an efficient method to target and electrophysiologically characterize intrinsic and synaptic properties of specific interneuron types. Combined with intracellular dye labeling, this approach can be extended with post-hoc morphological and immunocytochemical analysis, enabling systematic identification of recorded neurons. These methods can be tailored to suit a broad range of scientific questions regarding functional properties of diverse types of cortical neurons.

Cortical networks are controlled by a small, but diverse set of inhibitory interneurons. Functional investigation of interneurons therefore requires targeted recording and rigorous identification. Described here is a combined approach involving whole-cell recordings from single or synaptically-coupled pairs of neurons with intracellular labeling, post-hoc morphological and immunocytochemical analysis.

Morphologically-と神経化学的に特定し海馬介在ニューロンからホールセルパッチクランプ記録

GABAergic inhibitory interneurons play a central role within neuronal circuits of the brain. Interneurons comprise a small subset of the neuronal ...

Optogenetic Entrainment of Hippocampal Theta Oscillations in Behaving Mice

Extensive data on relationships of neural network oscillations to behavior and organization of neuronal discharge across brain regions call for new tools to selectively manipulate brain rhythms. Here we describe an approach combining projection-specific optogenetics with extracellular electrophysiology for high-fidelity control of hippocampal theta oscillations (5-10 Hz) in behaving mice. The specificity of the optogenetic entrainment is achieved by targeting channelrhodopsin-2 (ChR2) to the GABAergic population of medial septal cells, crucially involved in the generation of hippocampal theta oscillations, and a local synchronized activation of a subset of inhibitory septal afferents in the hippocampus. The efficacy of the optogenetic rhythm control is verified by a simultaneous monitoring of the local field potential (LFP) across lamina of the CA1 area and/or of neuronal discharge. Using this readily implementable preparation we show efficacy of various optogenetic stimulation protocols for induction of theta oscillations and for the manipulation of their frequency and regularity. Finally, a combination of the theta rhythm control with projection-specific inhibition addresses the readout of particular aspects of the hippocampal synchronization by efferent regions.

We describe the use of optogenetics and electrophysiological recordings for selective manipulations of hippocampal theta oscillations (5-10 Hz) in behaving mice. The efficacy of the rhythm entrainment is monitored using local field potential. A combination of opto- and pharmacogenetic inhibition addresses the efferent readout of hippocampal synchronization.

行動するマウスにおける海馬シータ振動の光同調

Extensive data on relationships of neural network oscillations to behavior and organization of neuronal discharge across brain regions call for new tools ...

Recording and Modulation of Epileptiform Activity in Rodent Brain Slices Coupled to Microelectrode Arrays

Temporal lobe epilepsy (TLE) is the most common partial complex epileptic syndrome and the least responsive to medications. Deep brain stimulation (DBS) is a promising approach when pharmacological treatment fails or neurosurgery is not recommended. Acute brain slices coupled to microelectrode arrays (MEAs) represent a valuable tool to study neuronal network interactions and their modulation by electrical stimulation. As compared to conventional extracellular recording techniques, they provide the added advantages of a greater number of observation points and a known inter-electrode distance, which allow studying the propagation path and speed of electrophysiological signals. However, tissue oxygenation may be greatly impaired during MEA recording, requiring a high perfusion rate, which comes at the cost of decreased signal-to-noise ratio and higher oscillations in the experimental temperature. Electrical stimulation further stresses the brain tissue, making it difficult to pursue prolonged recording/stimulation epochs. Moreover, electrical modulation of brain slice activity needs to target specific structures/pathways within the brain slice, requiring that electrode mapping be easily and quickly performed live during the experiment. Here, we illustrate how to perform the recording and electrical modulation of 4-aminopyridine (4AP)-induced epileptiform activity in rodent brain slices using planar MEAs. We show that the brain tissue obtained from mice outperforms rat brain tissue and is thus better suited for MEA experiments. This protocol guarantees the generation and maintenance of a stable epileptiform pattern that faithfully reproduces the electrophysiological features observed with conventional field potential recording, persists for several hours, and outlasts sustained electrical stimulation for prolonged epochs. Tissue viability throughout the experiment is achieved thanks to the use of a small-volume custom recording chamber allowing for laminar flow and quick solution exchange even at low (1 mL/min) perfusion rates. Quick MEA mapping for real-time monitoring and selection of stimulating electrodes is performed by a custom graphic user interface (GUI).

We illustrate how to perform recording and electrical modulation of 4-aminopyridine-induced epileptiform activity in rodent brain slices using microelectrode arrays. A custom recording chamber maintains tissue viability throughout prolonged experimental sessions. Live electrode mapping and selection of stimulating pairs are performed by a custom graphical user interface.

記録と電極アレイに結合された齧歯動物の脳スライスにおけるてんかん活動の変調

Temporal lobe epilepsy (TLE) is the most common partial complex epileptic syndrome and the least responsive to medications. Deep brain stimulation (DBS) ...

Recording Spatially Restricted Oscillations in the Hippocampus of Behaving Mice

The local field potential (LFP) emerges from ion movements across neural membranes. Since the voltage recorded by LFP electrodes reflects the summed electrical field of a large volume of brain tissue, extracting information about local activity is challenging. Studying neuronal microcircuits, however, requires a reliable distinction between truly local events and volume-conducted signals originating in distant brain areas. Current source density (CSD) analysis offers a solution for this problem by providing information about current sinks and sources in the vicinity of the electrodes. In brain areas with laminar cytoarchitecture such as the hippocampus, one-dimensional CSD can be obtained by estimating the second spatial derivative of the LFP. Here, we describe a method to record multilaminar LFPs using linear silicon probes implanted into the dorsal hippocampus. CSD traces are computed along individual shanks of the probe. This protocol thus describes a procedure to resolve spatially restricted neuronal network oscillations in the hippocampus of freely moving mice.

This protocol describes the recording of local field potentials with multi-shank linear silicon probes. Conversion of the signals using current source density analysis allows the reconstruction of local electrical activity in the mouse hippocampus. With this technique, spatially restricted brain oscillations can be studied in freely moving mice.

行動中マウスの海馬における振動を制限空間録音

The local field potential (LFP) emerges from ion movements across neural membranes. Since the voltage recorded by LFP electrodes reflects the summed ...

Selective Manipulation of Theta Oscillations in a Mouse Model

Source: Bender, F. et al., <a href="https://app.jove.com/v/57349">Optogenetic Entrainment of Hippocampal Theta Oscillations in Behaving Mice.</a> J. Vis. Exp (2018)
This video demonstrates a protocol to modulate theta oscillations in a genetically engineered mouse model with light-sensitive ion channels in its GABAergic medial septum neurons. By delivering light pulses to the hippocampus, the protocol aims to synchronize neuronal activity and generate theta oscillations, which are recorded as local field potentials.

マウスモデルにおけるシータ振動の選択的操作

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
PV-Cre ...

JoVE Encyclopedia of Experiments

Neurophysiology

Stimulation and Modulation Techniques

Optogenetic Control of Hippocampal Network Dynamics

Source: Manseau, F., et al. <a href="https://app.jove.com/v/55851">Tuning in the Hippocampal Theta Band In Vitro: Methodologies for Recording from the Isolated Rodent Septohippocampal Circuit.</a> J. Vis. Exp. (2017)
This video demonstrates the method to use optogenetics to modulate hippocampal network dynamics by activating genetically modified light-sensitive interneurons in a mouse brain slice, enhancing theta oscillation synchronization and studying neural circuit behavior.

海馬ネットワークダイナミクスの光遺伝学的制御

All procedures involving animal samples have been reviewed and approved by the appropriate animal ethical review committee.

	Optogenetic control in the ...

In Vitro Recording of Theta Oscillations in the Mouse Hippocampus

Source: Manseau, F., et.al. <a href="https://app.jove.com/v/55851">Tuning in the Hippocampal Theta Band In Vitro: Methodologies for Recording from the Isolated Rodent Septohippocampal Circuit.</a> J. Vis. Exp. (2017).
This video demonstrates the recording of rhythmic neuronal network activity, also known as theta oscillations, in an isolated hippocampus. Electrodes are inserted into various layers and spatial positions to detect and analyze theta oscillations. This shows the unique neuronal compositions and the connectivity of the neuronal network across different layers of the hippocampus.

海馬シータバンドのチューニング

要約

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