Name: optogenetic entrainment of hippocampal theta oscillations in behaving mice
Uploaded: 2018-06-29T13:30:00.000+00:00
Duration: 7 min 33 s
Description: Watch this Scientific Journal Video about Optogenetic Entrainment of Hippocampal Theta Oscillations in Behaving Mice at JoVE.com

原稿のデータ分析による専門家の助けのためのマリア ・ Gorbati とコメントのジェニファー Kupferman に感謝したいと思います。この作品は、ドイツ研究振興協会 (DFG; によって支えられました。Exc 257 NeuroCure、TK と AP;優先順位プログラム 1665、1799/1-1(2)、ハイゼンベルグ プログラム、AP 1799/2-1)、ドイツ ・ イスラエル科学研究開発 (GIF; 財団私-1326-421.13/2015 年、TK) と人類のフロンティア ・ サイエンス ・ プログラム (HFSP;RGY0076/2012、TK)。

著者が明らかに何もありません。

ここで私たちは同調行動中の動物の海馬シータを引き出す方法論が広くアクセスを発表しました。このアプローチは、情報処理や動作の θ の関数の研究に役立ちます。このメソッドの重要な側面が含まれます: (1) オプシンの選択と MS の軸索に chr2 ターゲット細胞海馬、継続的な刺激と LFP ように注入された光ファイバー線配列アセンブリの (2) 堅牢な光・電子機能マウス、シータ周波数で光?...

哺乳類の神経細胞の活動は、内および脳領域1,2,3,4間の情報伝達を支援するネットワークの振動によって調整されます。脳のリズムには、非常に遅い (< 0.8 Hz) 超高速 (> 200 Hz) 周波数まで至る振動が含まれます。証拠の大きいボディは、認知5,6,7,8,9,10 を含む多様な脳機能でネットワーク振動の関与をサポートしています、パーキンソン病やてんかんの13,14,15などの神経疾患と同様に、生得的な行動11,12 。同期の生理学的にもっともらしいモデルの開発と動作の因果関係を確立するため、ネットワークの振動の実験的操作メソッドが選択的な一時的は欠かせないため。
ネットワーク同期は多様な生物学的基質とイオン チャネルの分子の id、その動力学から興奮性とネットワーク接続性の神経に至るまでのプロセスを介する。発電機16が明らかにされている多くの脳のリズムが (例えば周波数、振幅) の異なる面が多いリズムの生物学的デザインは、明確な細胞のタイプとネットワークのダイナミクスによってもたらされます。例えば、主細胞の突起をターゲットと抑制性介在ニューロンは、周波数帯、脳領域17,18, シータ19、20、ガンマ20を含む最も重要な選手をします。,21日とリップル (140-200 Hz)22振動。ターンでは、離れた場所の細胞の位相同期は、堅牢なフィード フォワード信号錐体細胞の介在神経の発火をリセットによって保証されます。振動、同期ニューロン集団のサイズの重要なパラメーター測定 LFP 振動の振幅は密接な関係し、少なくとも高速振動2介在ニューロンに興奮性ドライブに依存します。対照的に、シータ、デルタなどの遅い振動は皮質視床23,24と海馬-内側中隔予測25,によって形成される長距離のリエントラント ループによって生成される26,27、それぞれ。このような回路の振動が信号伝搬遅延、興奮性の応答と周波数嗜好参加細胞28,29,30,の相互作用によってもたらされる31,32gaba 作動性パルブアルブミン (太陽光発電) から抑制性の投射-海馬25,33、海馬傍地域内嗅皮質26で介在する内側中隔 (MS) の細胞が陽性。内側側頭葉シータ振動の生成に不可欠。したがって、光遺伝学を用いたリアルタイム高精度ネットワーク振動・同期の神経の生理学的メカニズムを操作できます。
34,35,36,37,38 培養海馬と大脳皮質の振動やの研究のセル型固有の光遺伝学的操作を適用されています。生体内で30,39,40,41,42,43,44,45, 機能を含むガンマ5,12,36,46,47,48,49,50,の調査51,52とリップル振動40,53,54と睡眠のスピンドル、55,56。最近 PV Cre マウスの MS、海馬 θ リズムの生成のための重要な地域で Cre 依存 ChR2 ウイルスを表明しました。この準備を使用して、海馬シータ振動 (周波数と時間的安定性) の特徴は、海馬11MS の抑制性投射の光刺激によって制御されました。さらに、海馬海馬の抑制性投射のシータ振動への光刺激は目がさめている不動の中にシータを誘発しました。生じる気流 LFP および神経細胞の活動レベルでマウスで自発的なシータの θ リズムの表示プロパティです。
このプロトコルの主な機能: 海馬興奮性; 非特異的効果を回避しながら自発的なシータの重要な生理学的である抑制性経路の (1) の利用(2) 軸索、すなわち、非海馬 MS efferents; の直接影響を最小限に抑えるために特定の投影刺激(海馬海馬 θ リズムのダイナミクスとシータ振動の二国間の引き込みと直接干渉を最小限を確保 3) ローカルの θ リズム光刺激(4) パラメトリックな制御のシータ振動頻度および規則性;同調忠実高時間分解能 (5) 数量 LFP を使用して動物の行動に因果関係の定量的分析を有効にします。以来、この準備は、シータ世代25,30の脱抑制が海馬海馬のよく知られている役割を本質的に大文字、それはシータ振動動作マウスのいくつかのパラメーターのロバスト制御をできます。他より少ない調査経路および海馬海馬の細胞の種類はどこにいた研究操作38,39,47,49,50,51,52,53,54,55,56,57,58は θ リズムの機構をさらに明らかにします。

<table><tbody><tr><td>PV-Cre mice</td><td>The Jackson Laboratory</td><td>B6;129P2-Pvalbtm1(cre)Arbr/J</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;u&gt;Surgery&lt;/u&gt;</td><td></td><td></td><td></td></tr><tr><td>Stereotaxis</td><td>David Kopf Instruments, Tujunga, CA, USA</td><td>Model 963</td><td>Ultra Precise Small Animal Stereotaxic Instrument</td></tr><tr><td>Drill bits, 0.8 mm</td><td>Bijoutil, Allschwil, Switzerland</td><td>49080HM</td><td></td></tr><tr><td>0.01-1 ml syringe</td><td>Braun, Melsungen, Germany</td><td>9161406V</td><td></td></tr><tr><td>Sterican cannulas</td><td>Braun</td><td>26 G, 0.45x25 mm BL/LB</td><td></td></tr><tr><td>Fine and sharp scissors</td><td>Fine Science Tools Inc., Vancouver, Canada</td><td>14060-09</td><td></td></tr><tr><td>Forceps</td><td>Fine Science Tools Inc.</td><td>11210-10</td><td>Dumont AA - Epoxy Coated Forceps</td></tr><tr><td>Blunt stainless steel scissors</td><td>Fine Science Tools Inc.</td><td>14018-14</td><td></td></tr><tr><td>Soldering station</td><td>Weller Tools GmbH, Besigheim, Germany</td><td>WSD 81</td><td></td></tr><tr><td>Erythromycin</td><td>Rotexmedica GmbH, Trittau, Germany</td><td>PZN: 10823932</td><td>1g Powder for Solution for Infusion</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;u&gt;Optogenetics&lt;/u&gt;</td><td></td><td></td><td></td></tr><tr><td>Hamilton pump</td><td>PHD Ultra, Harvard Apparatus, Holliston, MA, USA</td><td>model 703008</td><td>PHD Ultra Syringe Pump with push/pull mechanism</td></tr><tr><td>Hamilton 5 &amp;micro;L Syringe, 26 gauge</td><td>PHD Ultra, Harvard Apparatus</td><td>Model 75 RN SYR</td><td></td></tr><tr><td>Hamilton 5 &amp;micro;L Plunger</td><td>PHD Ultra, Harvard Apparatus</td><td>Model 75 RN SYR</td><td></td></tr><tr><td>Tubing</td><td>Fisher Scientific, Pittsburgh, USA</td><td>PE 20</td><td>Inner diameter 0.38 mm (.015&quot;), Outer diameter 1.09 mm (.043&quot;)</td></tr><tr><td>Sterican cannulas</td><td>Braun, Melsungen, Germany</td><td>27 G, 25x0.40 mm, blunt</td><td></td></tr><tr><td>Precision drill/grinder</td><td>Proxxon, Wecker, Luxemburg</td><td>fbs 240/e</td><td></td></tr><tr><td>Cutting disks</td><td>Proxxon</td><td>NO 28812</td><td></td></tr><tr><td>Cre dependent channelrhodopsin</td><td>Penn Vector Core, Philadelphia, PA, USA</td><td>AV-1-18917P</td><td>Contruct name: AAV2/1.CAGGS.flex.ChR2.tdTomato, titer: 1.42x1013 vg/ml</td></tr><tr><td>Cam kinase dependent halorhodopsin</td><td>Penn Vector Core</td><td>AV-1-26971P</td><td>Construct name: eNpHR3.0, AAV2/1.CamKIIa.eNpHR3.0-EYFP.WPRE.hGH, titer: 2.08_1012 vg/ml</td></tr><tr><td>Multimode optic fiber</td><td>ThorLabs, Dachau, Germany</td><td>FG105LCA</td><td>0.22 NA, Low-OH, &amp;Oslash;105 &amp;micro;m Core, 400 - 2400 nm</td></tr><tr><td>Ceramic stick ferrule</td><td>Precision Fiber Products, Milpitas, CA, USA</td><td>CFLC126</td><td>Ceramic LC MM Ferrule, ID 126um</td></tr><tr><td>Polishing paper</td><td>Thorlabs</td><td>LF3D</td><td>6&quot; x 6&quot; Diamond Lapping (Polishing) Sheet</td></tr><tr><td>Power meter</td><td>Thorlabs</td><td>PM100D</td><td>Compact Power and Energy Meter Console, Digital 4&quot; LCD</td></tr><tr><td>Multimode fiber optic coupler</td><td>Thorlabs</td><td>FCMM50-50A-FC</td><td>1x2 MM Coupler, 50:50 Split Ratio, 50 &amp;micro;m GI Fibers, FC/PC</td></tr><tr><td>Fiberoptic patch cord</td><td>Thorlabs</td><td>FG105LCA CUSTOM-MUC</td><td>custom made, 3 m long, with protective tubing, Tubing: FT030, Connector 1: FC/PC, Connector 2: 1.25mm (LC) Ceramic Ferrule</td></tr><tr><td>Sleeve</td><td>Precision Fiber Products, Milpitas, CA, USA</td><td>ADAL1</td><td>Ceramic Split Mating Sleeve for &amp;Oslash;1.25 mm (LC/PC) Ferrules</td></tr><tr><td>473 nm DPSS laser</td><td>Laserglow Technologies, Toronto, ON, Canada</td><td>R471005FX</td><td>LRS-0473 Series</td></tr><tr><td>593 nm DPSS laser</td><td>Laserglow Technologies</td><td>R591005FX</td><td>LRS-0594 Series</td></tr><tr><td>MC_Stimulus II</td><td>Multichannel Systems, Reutlingen, Germany</td><td>STG 4004</td><td></td></tr><tr><td>Impedance conditioning module</td><td>Neural microTargeting worldwide, Bowdoin, USA</td><td>ICM</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;u&gt;Electrophysiology&lt;/u&gt;</td><td></td><td></td><td></td></tr><tr><td>Tungsten wires</td><td>California Fine Wire Company, Grover Beach, CA, USA</td><td>CFW0010954</td><td>40 &amp;micro;m, 99.95%</td></tr><tr><td>Capillary tubing</td><td>Optronics</td><td>1068150020</td><td>ID: 100.4 &amp;micro;m</td></tr><tr><td>Omnetics nanoconnector</td><td>Omnetics Connector Corporation, Minneapolis, USA</td><td>A79038-001</td><td></td></tr><tr><td>Screws</td><td>Bilaney, D&amp;uuml;sseldorf, Germany</td><td>00-96x1/16</td><td>stainless-steel</td></tr><tr><td>Silicone probe</td><td>NeuroNexus Technologies, Ann Arbor, MI, USA</td><td>B32</td><td></td></tr><tr><td>Headstage</td><td>Neuralynx, Bozeman, Montana USA</td><td>HS-8</td><td>miniature headstage unity gain preamplifiers</td></tr><tr><td>Silver conductive paint</td><td>Conrad electronics, Germany</td><td>530042</td><td></td></tr><tr><td>Liquid flux</td><td>Felder GMBH L&amp;ouml;ttechnik, Oberhausen, Germany</td><td>L&amp;ouml;t&amp;ouml;l ST</td><td>DIN EN 29454.1, 3.2.2.A (F-SW 11)</td></tr><tr><td>LED</td><td>Neuralynx</td><td>HS-LED-Red-omni-10V</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;u&gt;Software&lt;/u&gt;</td><td></td><td></td><td></td></tr><tr><td>MATLAB</td><td>Mathworks, Natick, MA, USA</td><td></td><td></td></tr><tr><td>MC_Stimulus software</td><td>Multichannel, Systems</td><td></td><td></td></tr><tr><td>Neurophysiological Data Manager</td><td></td><td></td><td>NDManager, http://neurosuite.sourceforge.net</td></tr><tr><td>Klusters</td><td></td><td></td><td>http://neurosuite.sourceforge.net, Hazan et al., 2006</td></tr><tr><td>Software of the recording system</td><td>Neuralynx</td><td>Cheetah</td><td>https://neuralynx.com/software/cheetah</td></tr><tr><td>Multi-channel data analysis software</td><td>Cambridge Electronic Design Limited, Cambridge, GB</td><td>Spike2</td><td></td></tr></tbody></table>

optogenetic entrainment of hippocampal theta oscillations in behaving mice

ニューラル ネットワークの動作する振動の関係や脳領域間の神経放電の組織に関する広範なデータを求める脳リズムを選択的に操作するための新しいツール。ここで特定の投影光遺伝学を組み合わせて動作マウスにおける海馬シータ振動 (5-10 Hz) の忠実度の高いコントロールの細胞電気生理学のアプローチについて述べる。重要な海馬シータ振動の生成に関与する内側中隔細胞の gaba 作動性の人口にターゲット チャネルロドプシン 2 (ChR2) により光同調の特異性を実現し、ローカル同期活性化海馬の抑制性中隔求心性神経のサブセット。ラミナ CA1 野のや神経細胞の放電の間でローカル フィールド ポテンシャル (LFP) の同時観測による光遺伝学的リズム制御の有効性が検証されます。この容易に実装可能な製剤を用いた各種光刺激プロトコル シータの誘導とその頻度と規則性の操作の有効性を示す.最後に、投影法固有の抑制と θ リズム制御の組み合わせは遠心性の地域によって海馬の同期の特定の面の読み出しをアドレスします。

セクション 1 で説明した MS の gaba 作動性セルに chr2 ターゲットは、図 2 aで示されています。背側海馬 CA1 領域の上移植は光ファイバー経由で MS gaba 作動性細胞の軸索の光遺伝学的刺激だけでなく、対側同側 (図 2 b) の刺激の周波数でシータをヘアレスします。半球 (図 2)。シータは、これの有効性は?...

Watch this Scientific Journal Video about Optogenetic Entrainment of Hippocampal Theta Oscillations in Behaving Mice at JoVE.com

Optogenetic Entrainment of Hippocampal Theta Oscillations in Behaving Mice

動作マウスにおける海馬シータ振動 (5-10 Hz) の選択を操作する光遺伝学、電気生理学的記録の使用について述べる。リズムの同調性は、ローカル フィールド ポテンシャルを使用して監視されます。光と薬理遺伝学的抑制の組み合わせは、海馬同期の遠心性の読み出しをアドレスします。

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 ...

optogenetic-entrainment-hippocampal-theta-oscillations-behaving

Universidad de Cartagena UDEC

Research

JoVE Journal

Neuroscience

11.6K ビュー.  Heinrich Heine University Düsseldorf. 動作マウスにおける海馬シータ振動 (5-10 Hz) の選択を操作する光遺伝学、電気生理学的記録の使用について述べる。リズムの同調性は、ローカル フィールド ポテンシャルを使用して監視されます。光と薬理遺伝学的抑制の組み合わせは、海馬同期の遠心性の読み出しをアドレスします。

ビデオ: Optogenetic Entrainment of Hippocampal Theta Oscillations in Behaving Mice

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

Optogenetic Manipulation of Neuronal Activity to Modulate Behavior in Freely Moving Mice

Optogenetic modulation of neuronal circuits in freely moving mice affects acute and long-term behavior. This method is able to perform manipulations of single neurons and region-specific transmitter release, up to whole neuronal circuitries in the central nervous system, and allows the direct measurement of behavioral outcomes. Neurons express optogenetic tools via an injection of viral vectors carrying the DNA of choice, such as Channelrhodopsin2 (ChR2). Light is brought into specific brain regions via chronic optical implants that terminate directly above the target region. After two weeks of recovery and proper tool-expression, mice can be repeatedly used for behavioral tests with optogenetic stimulation of the neurons of interest.
Optogenetic modulation has a high temporal and spatial resolution that can be accomplished with high cell specificity, compared to the commonly used methods such as chemical or electrical stimulation. The light does not harm neuronal tissue and can therefore be used for long-term experiments as well as for multiple behavioral experiments in one mouse. The possibilities of optogenetic tools are nearly unlimited and enable the activation or silencing of whole neurons, or even the manipulation of a specific receptor type by light.
The results of such behavioral experiments with integrated optogenetic stimulation directly visualizes changes in behavior caused by the manipulation. The behavior of the same animal without light stimulation as a baseline is a good control for induced changes. This allows a detailed overview of neuronal types or neurotransmitter systems involved in specific behaviors, such as anxiety. The plasticity of neuronal networks can also be investigated in great detail through long-term stimulation or behavioral observations after optical stimulation. Optogenetics will help to enlighten neuronal signaling in several kinds of neurological diseases.

With optogenetic manipulation of specific neuronal populations or brain regions, behavior can be modified with high temporal and spatial resolution in freely moving animals. By using different optogenetic tools in combination with chronically implanted optical fibers, a variety of neuronal modulations and behavioral testing can be performed.

自由に動くマウスの行動を調節する神経活動の光遺伝学的操作

Optogenetic modulation of neuronal circuits in freely moving mice affects acute and long-term behavior. This method is able to perform manipulations of ...

Generation of Local CA1 &#947; Oscillations by Tetanic Stimulation

Neuronal network oscillations are important features of brain activity in health and disease and can be modulated by a range of clinically used drugs. A protocol is provided to generate a model for studying CA1 &#947; oscillations (20 - 80 Hz). These &#947; oscillations are stable for at least 30 min&#160;and depend upon excitatory and inhibitory synaptic activity in addition to activation of pacemaker currents. Tetanically stimulated oscillations have a number of reproducible and easily quantifiable characteristics including spike count, oscillation duration, latency and frequency that report upon the network state. The advantages of the electrically stimulated oscillations include stability, reproducibility and episodic acquisition enabling robust characterization of network function. This model of CA1 &#947; oscillations can be used to study cellular mechanisms and to systematically investigate how neuronal network activity is altered in disease and by drugs. Disease state pharmacology can be readily incorporated by the use of brain slices from genetically modified or interventional animal models to enable selection of drugs that specifically target disease mechanisms.

Oscillations are fundamental network properties and are modulated by disease and drugs. Studying brain-slice oscillations allows characterization of isolated networks under controlled conditions. Protocols are provided for the preparation of acute brain slices for evoking CA1 &#947; oscillations.

テタヌス刺激による局所CA1γ振動の発生

Neuronal network oscillations are important features of brain activity in health and disease and can be modulated by a range of clinically used drugs. A ...

神経科学

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

This protocol outlines the procedures for preparing and recording from the isolated whole hippocampus, of WT and transgenic mice, along with recent improvements in methodologies and applications for the study of theta oscillations. A simple characterization of the isolated hippocampal preparation is presented whereby the relationship between internal hippocampal theta oscillators is examined together with the activity of pyramidal cells, and GABAergic interneurons, of the cornu ammonis-1 (CA1) and subiculum (SUB) areas. Overall, we show that the isolated hippocampus is capable of generating intrinsic theta oscillations in vitro and that rhythmicity generated within the hippocampus can be precisely manipulated by optogenetic stimulation of parvalbumin-positive (PV) interneurons. The in vitro isolated hippocampal preparation offers a unique opportunity to use simultaneous field and intracellular patch-clamp recordings from visually-identified neurons to better understand the mechanisms underlying theta rhythm generation.

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

Here, we present a protocol for recording rhythmic neuronal network theta and gamma oscillations from an isolated whole hippocampal preparation. We describe the experimental steps from extraction of the hippocampus to details of field, unitary and whole-cell patch clamp recordings as well as optogenetic pacing of the theta rhythm.

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 ...

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 ...

Optogenetic Manipulation of Neural Circuits During Monitoring Sleep/wakefulness States in Mice

In recent years, optogenetics has been widely used in many fields of neuroscientific research. In many cases, an opsin, such as channel rhodopsin 2 (ChR2), is expressed by a virus vector in a particular type of neuronal cells in various Cre-driver mice. Activation of these opsins is triggered by application of light pulses which are delivered by laser or LED through optic cables, and the effect of activation is observed with very high time resolution. Experimenters are able to acutely stimulate neurons while monitoring behavior or another physiological outcome in mice. Optogenetics can enable useful strategies to evaluate function of neuronal circuits in the regulation of sleep/wakefulness states in mice. Here we describe a technique for examining the effect of optogenetic manipulation of neurons with a specific chemical identity during electroencephalogram (EEG) and electromyogram (EMG) monitoring to evaluate the sleep stage of mice. As an example, we describe manipulation of GABAergic neurons in the bed nucleus of the stria terminalis (BNST). Acute optogenetic excitation of these neurons triggers a rapid transition to wakefulness when applied during NREM sleep. Optogenetic manipulation along with EEG/EMG recording can be applied to decipher the neuronal circuits that regulate sleep/wakefulness states.

Here, we describe methods of optogenetic manipulation of particular types of neurons during monitoring of sleep/wakefulness states in mice, presenting our recent work on the bed nucleus of the stria terminalis as an example.

マウスにおける睡眠/覚醒状態のモニタリング中の神経回路の光遺伝学的操作

In recent years, optogenetics has been widely used in many fields of neuroscientific research. In many cases, an opsin, such as channel rhodopsin 2 ...

Craniotomy Procedure for Visualizing Neuronal Activities in Hippocampus of Behaving Mice

Imaging neuronal activities at single-cell resolution in awake behaving animals is a very powerful approach for the investigation of neural circuit functions in systems neuroscience. However, high absorbance and scattering of light in mammalian tissue limit intravital imaging mostly to superficial brain regions, leaving deep-brain areas, such as the hippocampus, out of reach for optical microscopy. In this video, we show the preparation and implantation of the custom-made imaging window to enable chronic in vivo imaging of the dorsal hippocampal CA1 region in head-fixed behaving mice. The custom-made window is supplemented with an infusion cannula that allows targeted delivery of viral vectors and drugs to the imaging area. By combining this preparation with wide-field imaging, we performed a long-term recording of neuronal activity using a fluorescent calcium indicator from large subsets of neurons in behaving mice over several weeks. We also demonstrated the applicability of this preparation for voltage imaging with single-spike resolution. High-performance genetically encoded indicators of neuronal activity and scientific CMOS cameras allowed the recurrent visualization of subcellular morphological details of single neurons at high temporal resolution. We also discuss the advantages and potential limitations of the described method and its compatibility with other imaging techniques.

This article demonstrates the preparation of a custom-made imaging window supplemented with infusion cannula and its implantation onto the CA1 region of the hippocampus in mice.

マウスの振る舞いの海馬における神経活動を可視化するための頭蓋骨管制法

Imaging neuronal activities at single-cell resolution in awake behaving animals is a very powerful approach for the investigation of neural circuit ...

Selective Manipulation of Theta Oscillations in a Mouse Model

Source: Bender, F. et al., <a href="https://appjove.unicartagenaproxy.elogim.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

Implanting an Electrode Array and Optic Fiber into the Mouse Hippocampus

Source: Bender, F., et. al., <a href="https://appjove.unicartagenaproxy.elogim.com/v/57349">Optogenetic Entrainment of Hippocampal Theta Oscillations in Behaving Mice.</a> J. Vis. Exp. (2018)
This video demonstrates the stereotactic implantation of a wire electrode array and an optic fiber in the hippocampus of a mouse model.

電極アレイと光ファイバーのマウス海馬への埋め込み

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. ...

Optogenetic Control of Hippocampal Network Dynamics

Source: Manseau, F., et al. <a href="https://appjove.unicartagenaproxy.elogim.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 ...