アンドリュー ・ クリップスとレディング大学で生物ユニットに感謝したいと思います。この研究 BBSRC によって資金が供給された (許可番号: BB/K010123/1)。この作業に関連付けられているデータは Y.Z. (ying.zheng@reading.ac.uk) から自由に利用できます。

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H., Devor, A., Ulbert, I., Dale, A. M., Einevoll, G. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Current-source+density+estimation+based+on+inversion+of+electrostatic+forward+solution:+Effects+of+finite+extent+of+neuronal+activity+and+conductivity+discontinuities.">Current-source density estimation based on inversion of electrostatic forward solution: Effects of finite extent of neuronal activity and conductivity discontinuities.</a> J Neurosci Methods. 154 (1-2), 116-133 (2006).</li><li>Thomson, D. J., et al., Fitzgerald, W. 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=Multitaper+analysis+of+nonstationary+and+nonlinear+time+series+data.">Multitaper analysis of nonstationary and nonlinear time series data.</a> Nonlinear and Nonstationary Signal Processing. , 317-394 (2000).</li><li>Bae, J., Deshmukh, A., Song, Y., Riera, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Brain+Source+Imaging+in+Preclinical+Rat+Models+of+Focal+Epilepsy+using+High-Resolution+EEG+Recordings.">Brain Source Imaging in Preclinical Rat Models of Focal Epilepsy using High-Resolution EEG Recordings.</a> Journal of Visualized Experiments : JoVE. (100), e52700 (2015).</li><li>Baumann, S. B., Wozny, D. R., Kelly, S. K., Meno, F. 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著者が明らかに何もありません。

ウィスカー パッド刺激への応答のイソフルラン麻酔下ラットの共局在脳波と LFP 信号の同時記録の実験手順を説明しました。頭蓋骨に穴のバリと提携していた脳波クモ電極の開口部を通って大脳皮質に差し入れて電極は導電性で頭蓋骨に固定し、接着剤脳波は23を貼り付けます。イソフルランの管理に使用する鼻の円錐形は、簡単に、ウィスカー パッドに刺激電極を挿入可能性がありますので、変更されました。
脳波貼り付け貼り付けの追加のアプリケーションを必要とせず実験 1 日を通して優秀な電気伝導度を提供しながら安全に頭蓋骨にクモの電極を基板実装時有効であった。接着剤は非導べ電性頭蓋骨と電極間で実行する場合は、電極のインピー ダンスを増やすことができますと、頭蓋骨にクモ電極の周囲を修正する接着剤の望ましくない使用を交換してください。脳波ペーストには、図形のバリの穴回りにくいし、貧しい脳波の実験中に乾くことができます脳波ゲル上の利点の数があります。
ラットは、ファラデーケージの中に置かれた、環境による電気ノイズが大きく減衰されます。しかし、時々 神経信号はまだかなり騒々しかった。ほとんどの場合、これは安全に配置し、再調整する必要したがって、参照電極またはより多くの脳波によって引き起こされたペースト。別の一般的な問題は、振幅の誘発 LFP が小さいことでした。電極刺激電極によって活性化される皮質領域の中央に配置されていない可能性があります。再ローカル ニューロンへより多くのダメージの原因、微小電極を挿入する代わりに我々 は通常、LFP の合理的な振幅までウィスカー パッドで刺激電極の位置を調整 (> 3 mV) 観察することができます。
技術の制限の 1 つは、6 mm の直径を持つクモの電極の空間解像度が低いです。これはラットの頭蓋骨のサイズと比較して大きいです。残念ながら、ここで使用されるクモ電極は最小の購入に利用できます。2-4 mm スパイダー電極の直径を削減することが望ましいこと、脳波、脳波の信号と、顆粒とを比較しての空間の特異性をこうして増加 LFP 信号曖昧さの少ない。
プロトコルのいくつかの重要な手順には、特別な注意が必要があります。最初のバリの穴を介して電極の挿入です。硬膜はそのままそれ以外の場合、カーソル位置の精度は重要です。電極の先端に僅かな抵抗は通常電極が正しく配置されていないを意味します。それが発生する必要があります、位置調整、再挿入します。2 番目は、ラットの鼻の円錐形の位置です。イソフルランは円錐形からの脱出としてそれは緩すぎるなりません。それもしないでくださいきつすぎるとラットの鼻の穴を妨げるし、呼吸困難を引き起こすことができます。特別な注意は、脳波の振幅が小さくなっていることを確認するも必要です (通常 5 ~ 10 倍小さい) LFP 最上位チャネル記録より。彼らが類似している場合に直接または間接的な接触電極脳波プローブが来ていることを示すです。間接的な接触は通常時の穴を満たす脳脊髄液 (CSF) を掘削頭蓋骨の。髄液の電気伝導度は通常 100 回頭蓋骨24,25 です。したがって、バリ穴の中の脳脊髄液のレベルが十分に高い場合、それを作ることができますスパイダー電極と接触します。これを避けるためには、超吸収性綿スポンジ吸収槍などとの穴を頻繁掃除必要があります。
バリの穴の効果 (直径 < 2 mm) 脳波に頭蓋骨の穴を囲む記録 ipsi 水平バレル皮質上にドクロをそのままに別のクモの電極配置をする二国間の脳波を比較することによって調べた。図 9 と図 10に示すように有意水準 0.05 で有意に効果が示唆されました。脳波のペーストは、頭蓋骨、電極が貼り付け] を押されたどのようにしっかりと頭蓋の脳波ペーストの空間的な広がりとの接触がいかにうまく、脳波の振幅に影響を与えるその他の要因が含まれます。
また、ここで説明されているプロトコルが、人間の脳波の研究で使用される脳波の頭皮から異なる頭蓋骨脳波記録ことに注意する価値があります。頭皮抵抗やさらに脳波の信号対雑音比を低減する低域通過フィルターのような役割を果たします。
最後に、ERP のダイナミクスと皮質層にわたって誘発 LFP の比較は、体性感覚誘発電位より良い粒状より皮質の顆粒層とではレイヤー LFP を反映するをお勧めします。これは以前仕事6ERP の最初のセグメント (P1) は興奮性シナプス電流発生の顆粒層でその後減少 (流入から生じる戻り現在に関連しているを示すと一致しています。N1) erp に関連する視床皮質への求心性の層 II/III および/またはフィード フォワード信号の到着が遅れて深い皮質層から。結論として、脳波/LFP の同時録音することができます脳波、神経の起源に関する理解を深めるし、皮質層全体の神経信号の面で脳波の数学的モデリングを容易にします。

一般に電極を介して主に記録の LFPs がローカル錐体神経集団1,2,3の同期興奮性と抑制性シナプスの活動の加重の合計を反映しています。,4. 私たちの最近の研究が示した LFP 信号のプロファイルが興奮と抑制の5,6のコンポーネントに分けることができます。ただし、LFP 侵襲で通常量を測定したら、人間の脳のほとんどの研究にもない適しています。
その一方で、脳波は、脳の電気的活動を測定する非侵襲的な手法です。特定の種類、てんかんなどの神経疾患のための診断ツールとして、人間の認知科学的研究の研究ツールとして広く使用されます。その人気にもかかわらず、脳波の主要な制限は、基になる神経信号7,8,9面で正確にその時間プロファイルを解釈することができないです。
ますます、脳波の数学的モデルは、脳機能10、11,12,13,14,15の理解を高めるために開発されています。既存の脳波モデルのほとんどは、フィッティング中、自発脳波データに出力予測モデルの周波数ドメインの特性に基づいて開発されて、非常に少数の脳波モデルは現実的な感覚誘発電位を生成できます。このコンテキストで LFP と脳波の同時録音は脳波のより正確な数学モデルを開発するため重要な洞察力と制約が提供されます。
さらに脳波の神経の起源を探検する同時録音のこの必要性に対処するため麻酔下ラットの大脳皮質脳波とマルチ-層流 LFP 信号を同時に記録する手法を開発しました。セットアップは、以前同時脳波/LFP 研究実施、霊長類16,17に似ています。さらに脳波感覚の不在で二国間 EEG 録音は (すなわちバリの穴、そのまま他の半球の 1 つの半球) を比較することによって、ホールの周りで頭蓋骨に穴のバリの効果を調べた刺激。脳波/LFP の同時録音が行なわれること単にかつ効果的に、頭蓋骨のバリの穴から小さな脳波信号の歪みとを示した。

<table>
	<tbody>
		<tr>
			<td>Female Lister Hood rats</td>
			<td>Charles Rivers</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Spider electrode</td>
			<td>Unimed Electrode Supplies Ltd</td>
			<td>SCS24-426</td>
			<td></td>
		</tr>
		<tr>
			<td>EEG paste: Ten20</td>
			<td>Unimed Electrode Supplies Ltd</td>
			<td>10-20-S</td>
			<td></td>
		</tr>
		<tr>
			<td>Stereotaxic holder with dual micromanipulator arms: Dual Manipulator Stereotaxic Frame with 18&#176; Ear Bars</td>
			<td>WPI (World Precision Instruments)</td>
			<td>502603</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>National Vet Services Limited</td>
			<td>50878</td>
			<td></td>
		</tr>
		<tr>
			<td>Hard plastic nose cone: Anasthesia Gas Mask for Rat</td>
			<td>WPI</td>
			<td>502054</td>
			<td></td>
		</tr>
		<tr>
			<td>Small animal isoflurane anaesthetic system</td>
			<td>WPI</td>
			<td>EZ-B800A</td>
			<td></td>
		</tr>
		<tr>
			<td>Thermostatic heating pad: Rat Blanket System 230V</td>
			<td>Harvard Apparatus UK</td>
			<td>50-7221-F</td>
			<td></td>
		</tr>
		<tr>
			<td>Ophthalmic ointment: Optixcare eye lube</td>
			<td>Viovet</td>
			<td>203865</td>
			<td></td>
		</tr>
		<tr>
			<td>Lidocaine Hydrochloride (Injection 2%)</td>
			<td>Larkmead Vets</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Jacquette Scaler #1SSE, 18cm, Hollow</td>
			<td>WPI</td>
			<td>503421</td>
			<td></td>
		</tr>
		<tr>
			<td>Serrated and curved dissecting forceps</td>
			<td>WPI</td>
			<td>15915</td>
			<td></td>
		</tr>
		<tr>
			<td>Braided silk, non-absorbable suture: Mersilk Suture W502H</td>
			<td>National Vet Services Limited</td>
			<td>153746</td>
			<td></td>
		</tr>
		<tr>
			<td>Dental drill: BONE MICRO DRILL SYST 230 VAC</td>
			<td>Harvard Apparatus UK</td>
			<td>72-4860</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile Saline: Sodium chloride 0.9%</td>
			<td>Animalcare Ltd</td>
			<td>14K26BT</td>
			<td></td>
		</tr>
		<tr>
			<td>Drill bit #4 : Ball Mill, Carbide, #4</td>
			<td>Harvard Apparatus UK</td>
			<td>72-4958</td>
			<td></td>
		</tr>
		<tr>
			<td>Drill bit #4 : Ball Mill, Carbide, #1/4</td>
			<td>Harvard Apparatus UK</td>
			<td>72-4962</td>
			<td></td>
		</tr>
		<tr>
			<td>Faraday cage</td>
			<td>Newport Corporation</td>
			<td>VIS-FDC-3600</td>
			<td></td>
		</tr>
		<tr>
			<td>Vibration isolation workstation: Vision IsoStation</td>
			<td>Newport Corporation</td>
			<td>M-VIS3660-RG4-325A</td>
			<td></td>
		</tr>
		<tr>
			<td>Oximeter Control Unit and sensor: MouseOxPlus, Starr Life Sciences Corp.</td>
			<td>WPI</td>
			<td>O15001</td>
			<td></td>
		</tr>
		<tr>
			<td>Transparent soft nose cone: Microflex Non-Rebreathing Unit with a Rat Nosecone</td>
			<td>WPI</td>
			<td>EZ-103A</td>
			<td></td>
		</tr>
		<tr>
			<td>Stainless steel stimulating electrodes</td>
			<td>PlasticsOne</td>
			<td>E363/1/SPC</td>
			<td></td>
		</tr>
		<tr>
			<td>Isolated current stimulator</td>
			<td>Made in House</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>16-channel micro-electrode, 100 &#956;m spacing, area of each site 177 &#956;m2</td>
			<td>NeuroNexus</td>
			<td>A1x16-10mm-100-177-A16</td>
			<td></td>
		</tr>
		<tr>
			<td>16-channel acute headstage</td>
			<td>Tucker David Technologies Inc., TDT</td>
			<td>RA16AC-Z</td>
			<td></td>
		</tr>
		<tr>
			<td>Pre-Amplifier: Z-Series 64-Channel Neuro-Digitizing Preamp</td>
			<td>TDT</td>
			<td>PZ5-64</td>
			<td></td>
		</tr>
		<tr>
			<td>Passive signal splitter: 32-Channel Splitter Box for PZ5</td>
			<td>TDT</td>
			<td>S-BOX_PZ5</td>
			<td></td>
		</tr>
		<tr>
			<td>Data acquisition unit: RZ2 BioAmp Processor. Z-Series 4-DSP ultra high performance processor</td>
			<td>TDT</td>
			<td>RZ2-4</td>
			<td></td>
		</tr>
		<tr>
			<td>Software for Neurophysiology: OpenEX</td>
			<td>TDT</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Matlab</td>
			<td>MathWorks</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Absorption spears</td>
			<td>Fine Sicence Tools</td>
			<td>18105-01</td>
			<td></td>
		</tr>
	</tbody>
</table>

concurrent recording of co-localized electroencephalography and local field potential in rodent

脳波 (EEG) は非侵襲的手法として普及し、脳の神経活動を記録するため、脳波の神経新生の私達の理解はまだ非常に限られました。マルチ-層流電極を介して記録局所電界電位 (LFPs) は、大脳皮質の異なる皮質層にわたって同時の神経活動の詳細なアカウントを提供できるが侵襲的な手法。前臨床モデルで LFP と脳波の測定値を結合することができます大幅に脳波信号の生成に関与する神経機構の理解を強化し、脳波のより現実的な生物学的に正確な数学的モデルの導出を容易にします。同時と共局在脳波と麻酔の齧歯類でマルチ層流の LFP 信号を取得するための簡単な手順を次に示します。また脳波信号が微小電極の挿入のため頭蓋骨の穴のバリの影響を受けますかどうかを調べた。バリの穴では、脳波のごくわずかな影響が示唆されました。

該当する時系列を取得 4 ラットからのデータを平均しました。誘発脳波応答の振幅として知られている、事象関連電位 (ERP) は LFP のそれより普通大いに小さく。図 6は、それぞれ平均 ERP と顆粒、粒状で LFP とバレル野のではレイヤーを示します。各プロットのエラー バンドは、対応する標準エラーです。ERP が 10 回誘発 LFP より小さい約こと見ることができます。
<img alt="Figure 6" class="xfigimg" src="/files/ftp_upload/56447/56447fig6.jpg"> 図 6: 意味 (n = 4) では LFP、顆粒、粒状、ERP の神経信号します。シャドウは、標準エラーを示します。<a href="//cloudflare-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56447/56447fig6large.jpg" target="_blank">この図の拡大版を表示するのにはここをクリックしてください</a>。
ERP と LFP のダイナミクスの比較は図 7のとおりです。ERP と顆粒図 7Aの LFP の直接の重ね合わせは、振幅の違い神経信号のこれらの 2 つのタイプの順序を示しています。時空間ダイナミクスを比較するには、ERP と LFP はその負のピーク振幅に対して正規化されます。図 7 b 7 Cは、正規化された ERP 重ね書き正規化された顆粒 LFP と正規化された粒状 LFP それぞれを表示します。
それは、P1, N1 向け ERP のピークは顆粒層で LFP の対応するピークより遅れて図 7Bから見ることができます。しかし、これら 2 つの神経信号の一時的なプロファイルは、同様に、P1 と N1 の前します。一方、ERP の一時的なプロファイルは顆粒 (層 IV バレル皮質) の著しく異なる LFP (図 7)。重要なは彼らは ERP から成っていた主に 2 つのピーク逆極性を持つに対し、単一負のピーク (皮質層 IV の主要なシンクを反映して)、によって支配される粒状 LFP と互いのミラー イメージではありません。
<img alt="Figure 7" class="xfigimg" src="/files/ftp_upload/56447/56447fig7.jpg"> 図 7: ERP と LFP のダイナミクスの比較。(A) ERP (実線) 顆粒 LFP (破線) 重ね書き。シャドウは、標準エラーを示します。(B) から構成される正規化された ERP (実線) で、正規化された顆粒 LFP (破線) 重ね書き。(C) から構成される正規化された ERP (実線) で、正規化された粒状 LFP (破線) 重ね書き。<a href="//cloudflare-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56447/56447fig7large.jpg" target="_blank">この図の拡大版を表示するのにはここをクリックしてください</a>。
それに穴のバリ、頭蓋骨にクモ電極を介して ERP 信号を測定しました。穴の脳波に及ぼす影響を調べるためには、別のクモの電極は ipsi 水平バレル皮質上のドクロをそのままに置かれました。脳波ペーストの量を調整することによって 2 つくも電極のインピー ダンスを大きさに匹敵したように注意されました。4 つのラットからのデータ (ない同じラット使用された上記) をご紹介します。
図 8 図 8 aに表示される 100 s データと長方形フレームのデータ同時の休憩状態 1 匹のラットの両方の電極から脳波の録音を示しています (20 s)図 8 bで展開されています。2 つの脳波信号主共変、振幅の類似範囲内。図 9は、周波数軸上線形スケールを使用して一番上の行と下の行より低い周波数範囲の拡大された眺めを提供するために周波数軸の対数スケールの使用 4 ラットの psd ファイルを示します。図 9からがしないよう科目間で psd ファイルで一貫したバイアスであります。これは図 10に示すように、5 つの周波数帯で平均化された psd ファイルの正規化の違いに 1 サンプルの t テストを実行することによって確認されました。これらの周波数の正規化された PSD 違いのどれもはゼロから有意 (p = 0.32、0.46、0.85、0.69、0.97、それぞれ)。
<img alt="Figure 8" class="xfigimg" src="/files/ftp_upload/56447/56447fig8.jpg"> 図 8: 二国間脳波。休眠状態の頭蓋骨 (黒) と (グレー) そのままの頭蓋骨と反対側の半球に同時脳波記録でバリの穴との間に (A) の頭蓋骨脳波(B) 展開は、(A) の長方形フレーム内の波形の表示します。<a href="//cloudflare-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56447/56447fig8large.jpg" target="_blank">この図の拡大版を表示するのにはここをクリックしてください</a>。
<img alt="Figure 9" class="xfigimg" src="/files/ftp_upload/56447/56447fig9.jpg"> 図 9: コントラ - (青) と ipsi 側 (赤) 脳波パワー スペクトル密度 (PSD).各列には、1 匹のラットの psd ファイルが表示されます。トップのパネルは、最下部のパネルは、可視化することより低い周波数範囲で psd ファイルを許可する対数周波数スケールを使用しながら、直線周波数スケールを使用します。<a href="//cloudflare-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56447/56447fig9large.jpg" target="_blank">この図の拡大版を表示するのにはここをクリックしてください</a>。
<img alt="Figure 10" class="xfigimg" src="/files/ftp_upload/56447/56447fig10.jpg"> 図 10: グループの分析。コントラ - と 5 つの周波数帯を持つ ipsi 横 PSD の違いを正規化: デルタ、シータ、アルファ、ベータ、ガンマ。各バーは、エラー バーとして表示される標準誤差と周波数帯域内平均正規化の違いを示しています。<a href="//cloudflare-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56447/56447fig10large.jpg" target="_blank">この図の拡大版を表示するのにはここをクリックしてください</a>。

Watch this Scientific Journal Video about Concurrent Recording of Co-localized Electroencephalography and Local Field Potential in Rodent at JoVE.com

Concurrent Recording of Co-localized Electroencephalography and Local Field Potential in Rodent

このプロトコルでは、共局在脳波 (EEG) や麻酔下のラットにおける潜在的なマルチ-層流のローカル フィールドの同時録音のための簡単な方法について説明します。脳波信号の歪みを生成する微小電極の挿入のため頭蓋骨の穴のバリが表示されます。

共局在脳波および齧歯動物のローカル フィールド電位の同時記録

Although electroencephalography (EEG) is widely used as a non-invasive technique for recording neural activities of the brain, our understanding of the ...

concurrent-recording-co-localized-electroencephalography-local-field

Nanyang Technological University

Research

JoVE Journal

Neuroscience

12.2K ビュー.  University of Reading. このプロトコルでは、共局在脳波 (EEG) や麻酔下のラットにおける潜在的なマルチ-層流のローカル フィールドの同時録音のための簡単な方法について説明します。脳波信号の歪みを生成する微小電極の挿入のため頭蓋骨の穴のバリが表示されます。

ビデオ: Concurrent Recording of Co-localized Electroencephalography and Local Field Potential in Rodent

Systemic and Local Drug Delivery for Treating Diseases of the Central Nervous System in Rodent Models

Thorough preclinical testing of central nervous system (CNS) therapeutics includes a consideration of routes of administration and agent biodistribution in assessing therapeutic efficacy. Between the two major classifications of administration, local vs. systemic, systemic delivery approaches are often preferred due to ease of administration. However, systemic delivery may result in suboptimal drug concentration being achieved in the CNS, and lead to erroneous conclusions regarding agent efficacy. Local drug delivery methods are more invasive, but may be necessary to achieve therapeutic CNS drug levels. Here, we demonstrate proper technique for three routes of systemic drug delivery: intravenous injection, intraperitoneal injection, and oral gavage. In addition, we show a method for local delivery to the brain: convection-enhanced delivery (CED). The use of fluorescently-labeled compounds is included for in vivo imaging and verification of proper drug administration. The methods are presented using murine models, but can easily be adapted for use in rats.

Thorough preclinical testing of drugs that act in the central nervous system often involves assessing and comparing drug biodistribution in association with specific routes of administration. Here, three commonly used methods of systemic delivery (intravenous, intraperitoneal, and oral) as well as a method for local delivery (convection-enhanced delivery) are demonstrated in mice.

齧歯類モデルで中枢神経系の疾患を治療するための全身および局所薬物送達

Thorough preclinical testing of central nervous system (CNS) therapeutics includes a consideration of routes of administration and agent biodistribution ...

神経科学

Examining Local Network Processing using Multi-contact Laminar Electrode Recording

Cortical layers are ubiquitous structures throughout neocortex1-4 that consist of highly recurrent local networks. In recent years, significant progress has been made in our understanding of differences in response properties of neurons in different cortical layers5-8, yet there is still a great deal left to learn about whether and how neuronal populations encode information in a laminar-specific manner.
Existing multi-electrode array techniques, although informative for measuring responses across many millimeters of cortical space along the cortical surface, are unsuitable to approach the issue of laminar cortical circuits. Here, we present our method for setting up and recording individual neurons and local field potentials (LFPs) across cortical layers of primary visual cortex (V1) utilizing multi-contact laminar electrodes (Figure 1; Plextrode U-Probe, Plexon Inc).
The methods included are recording device construction, identification of cortical layers, and identification of receptive fields of individual neurons. To identify cortical layers, we measure the evoked response potentials (ERPs) of the LFP time-series using full-field flashed stimuli. We then perform current-source density (CSD) analysis to identify the polarity inversion accompanied by the sink-source configuration at the base of layer 4 (the sink is inside layer 4, subsequently referred to as granular layer9-12). Current-source density is useful because it provides an index of the location, direction, and density of transmembrane current flow, allowing us to accurately position electrodes to record from all layers in a single penetration6, 11, 12.

A fundamental issue in our understanding of cortical circuitry is how networks in different cortical layers encode sensory information. Here we describe electrophysiological techniques utilizing multi-contact laminar electrodes to record single-units and local field potentials and present analyses to identify cortical layers.

マルチ連絡ラミナーの電極の録音を使用してローカルネットワークの処理を検討

Cortical layers are ubiquitous structures throughout neocortex1-4 that consist of highly recurrent local networks. In recent years, significant progress ...

Simultaneous Electroencephalography, Real-time Measurement of Lactate Concentration and Optogenetic Manipulation of Neuronal Activity in the Rodent Cerebral Cortex

Although the brain represents less than 5% of the body by mass, it utilizes approximately one quarter of the glucose used by the body at rest1. The function of non rapid eye movement sleep (NREMS), the largest portion of sleep by time, is uncertain. However, one salient feature of NREMS is a significant reduction in the rate of cerebral glucose utilization relative to wakefulness2-4. This and other findings have led to the widely held belief that sleep serves a function related to cerebral metabolism. Yet, the mechanisms underlying the reduction in cerebral glucose metabolism during NREMS remain to be elucidated.
	One phenomenon associated with NREMS that might impact cerebral metabolic rate is the occurrence of slow waves, oscillations at frequencies less than 4 Hz, in the electroencephalogram5,6. These slow waves detected at the level of the skull or cerebral cortical surface reflect the oscillations of underlying neurons between a depolarized/up state and a hyperpolarized/down state7. During the down state, cells do not undergo action potentials for intervals of up to several hundred milliseconds. Restoration of ionic concentration gradients subsequent to action potentials represents a significant metabolic load on the cell8; absence of action potentials during down states associated with NREMS may contribute to reduced metabolism relative to wake.
	Two technical challenges had to be addressed in order for this hypothetical relationship to be tested. First, it was necessary to measure cerebral glycolytic metabolism with a temporal resolution reflective of the dynamics of the cerebral EEG (that is, over seconds rather than minutes). To do so, we measured the concentration of lactate, the product of aerobic glycolysis, and therefore a readout of the rate of glucose metabolism in the brains of mice. Lactate was measured using a lactate oxidase based real time sensor embedded in the frontal cortex. The sensing mechanism consists of a platinum-iridium electrode surrounded by a layer of lactate oxidase molecules. Metabolism of lactate by lactate oxidase produces hydrogen peroxide, which produces a current in the platinum-iridium electrode. So a ramping up of cerebral glycolysis provides an increase in the concentration of substrate for lactate oxidase, which then is reflected in increased current at the sensing electrode. It was additionally necessary to measure these variables while manipulating the excitability of the cerebral cortex, in order to isolate this variable from other facets of NREMS.
	We devised an experimental system for simultaneous measurement of neuronal activity via the elecetroencephalogram, measurement of glycolytic flux via a lactate biosensor, and manipulation of cerebral cortical neuronal activity via optogenetic activation of pyramidal neurons. We have utilized this system to document the relationship between sleep-related electroencephalographic waveforms and the moment-to-moment dynamics of lactate concentration in the cerebral cortex. The protocol may be useful for any individual interested in studying, in freely behaving rodents, the relationship between neuronal activity measured at the electroencephalographic level and cellular energetics within the brain.

A procedure is described for manipulating the activity of cerebral cortical pyramidal neurons optogenetically while the electroencephalogram, electromyogram, and cerebral lactate concentration are monitored. Experimental recordings are performed on cable-tethered mice while they undergo spontaneous sleep/wake cycles. Optogenetic equipment is assembled in our laboratory; recording equipment is commercially available.

同時脳波、リアルタイム乳酸濃度の測定と齧歯類大脳皮質ニューロン活動のOptogeneticマニピュレーション

Although the brain represents less than 5% of the body  by mass, it utilizes approximately one quarter of the glucose used by the body  at rest1. The ...

Intracellular Recording, Sensory Field Mapping, and Culturing Identified Neurons in the Leech, Hirudo medicinalis

The freshwater leech, Hirudo medicinalis, is a versatile model organism that has been used to address scientific questions in the fields of neurophysiology, neuroethology, and developmental biology. The goal of this report is to consolidate experimental techniques from the leech system into a single article that will be of use to physiologists with expertise in other nervous system preparations, or to biology students with little or no electrophysiology experience. We demonstrate how to dissect the leech for recording intracellularly from identified neural circuits in the ganglion. Next we show how individual cells of known function can be removed from the ganglion to be cultured in a Petri dish, and how to record from those neurons in culture. Then we demonstrate how to prepare a patch of innervated skin to be used for mapping sensory or motor fields. These leech preparations are still widely used to address basic electrical properties of neural networks, behavior, synaptogenesis, and development. They are also an appropriate training module for neuroscience or physiology teaching laboratories.

Intracellular Recording, Sensory Field Mapping, and Culturing Identified Neurons in the Leech, Hirudo medicinalis

This article describes three nervous system preparations using leeches: intracellular recording from neurons in ventral ganglia, culturing neurons from ventral ganglia, and recording from a patch of innervated skin to map sensory fields.

細胞内記録、感覚野のマッピング、および培養は、リーチのニューロンを同定しヒルドmedicinalis</i

The freshwater leech, Hirudo medicinalis, is a versatile model organism that has been used to address scientific questions in the fields of ...

Measuring Spinal Presynaptic Inhibition in Mice By Dorsal Root Potential Recording In Vivo

Presynaptic inhibition is one of the most powerful inhibitory mechanisms in the spinal cord. The underlying physiological mechanism is a depolarization of primary afferent fibers mediated by GABAergic axo-axonal synapses (primary afferent depolarization). The strength of primary afferent depolarization can be measured by recording of volume-conducted potentials at the dorsal root (dorsal root potentials, DRP). Pathological changes of presynaptic inhibition are crucial in the abnormal central processing of certain pain conditions and in some disorders of motor hyperexcitability. Here, we describe a method of recording DRP in vivo in mice. The preparation of spinal cord dorsal roots in the anesthetized animal and the recording procedure using suction electrodes are explained. This method allows measuring GABAergic DRP and thereby estimating spinal presynaptic inhibition in the living mouse. In combination with transgenic mouse models, DRP recording may serve as a powerful tool to investigate disease-associated spinal pathophysiology. In vivo recording has several advantages compared to ex vivo isolated spinal cord preparations, e.g. the possibility of simultaneous recording or manipulation of supraspinal networks and induction of DRP by stimulation of peripheral nerves.

Measuring Spinal Presynaptic Inhibition in Mice By Dorsal Root Potential Recording In Vivo

GABAergic presynaptic inhibition is a powerful inhibitory mechanism in the spinal cord important for motor and sensory signal integration in spinal cord networks. Underlying primary afferent depolarization can be measured by recording of dorsal root potentials (DRP). Here we demonstrate a method of in vivo recording of DRP in mice.

後根潜在的記録によりマウスにおける脊髄シナプス前抑制を測定する<em&gt;インビボ</em

Presynaptic inhibition is one of the most powerful inhibitory mechanisms in the spinal cord. The underlying physiological mechanism is a depolarization of ...

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

Here we describe a method for the construction of a single-use &#8220;injectrode&#8221; using commercially accessible and affordable parts. A probing system was developed that allows for the injection of a drug while recording electrophysiological signals from the affected neuronal population. This method provides a simple and economical alternative to commercial solutions. A glass pipette was modified by combining it with a hypodermic needle and a silver filament. The injectrode is attached to commercial microsyringe pump for drug delivery. This results in a technique that provides real-time pharmacodynamics feedback through multi-unit extracellular signals originating from the site of drug delivery. As a proof of concept, we recorded neuronal activity from the superior colliculus elicited by flashes of light in rats, concomitantly with delivery of drugs through the injectrode. The injectrode recording capacity permits the functional characterization of the injection site favoring precise control over the localization of drug delivery. Application of this method also extends far beyond what is demonstrated here, as the choice of chemical substance loaded into the injectrode is vast, including tracing markers for anatomic experiments.

Here, we craft a glass pipette with dual functions: inhibition of deep brain structures by microinjections of drugs and real-time monitoring of their effects through simultaneous electrophysiological recordings.

同時電気生理学的記録およびげっ歯類の脳における阻害剤のマイクロ注射

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

Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats

Converging evidence shows that many neuropsychiatric diseases should be understood as disorders of large-scale neuronal networks. To better understand the pathophysiological basis of these diseases, it is necessary to precisely characterize in which way the processing of information is disturbed between the different neuronal parts of the circuit. Using extracellular in vivo electrophysiological recordings, it is possible to accurately delineate neuronal activity within a neuronal network. The application of this method has several advantages over alternative techniques, e.g., functional magnetic resonance imaging and calcium imaging, as it allows a unique temporal and spatial resolution and does not rely on genetically engineered organisms. However, the use of extracellular in vivo recordings is limited since it is an invasive technique that cannot be universally applied. In this article, a simple and easy to use method is presented with which it is possible to simultaneously record extracellular potentials such as local field potentials and multiunit activity at multiple sites of a network. It is detailed how a precise targeting of subcortical nuclei can be achieved using a combination of stereotactic surgery and online analysis of multi-unit recordings. Thus, it is demonstrated, how a complete network such as the hyperdirect cortico-basal ganglia loop can be studied in anesthetized animals in vivo.

Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats

In this study, the methodology is presented on how to perform multi-site in vivo electrophysiological recordings from the hyperdirect pathway under urethane anesthesia.

急性<em&gt;インビボ</em&gt;麻酔ラットにおけるHyperdirect経路からの局所場ポテンシャルとマルチユニット活性の電気生理学的記録

Converging evidence shows that many neuropsychiatric diseases should be understood as disorders of large-scale neuronal networks. To better understand the ...

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) ...

Evaluation of Hemisphere Lateralization with Bilateral Local Field Potential Recording in Secondary Motor Cortex of Mice

This article demonstrates complete, detailed procedures for both in vivo bilateral recording and analysis of local field potential (LFP) in the cortical areas of mice, which are useful for evaluating possible laterality deficits, as well as for assessing brain connectivity and coupling of neural network activities in rodents. The pathological mechanisms underlying Alzheimer&#39;s disease (AD), a common neurodegenerative disease, remain largely unknown. Altered brain laterality has been demonstrated in aging people, but whether or not abnormal lateralization is one of the early signs of AD has not been determined. To investigate this, we recorded bilateral LFPs in 3-5-month-old AD model mice, APP/PS1, together with littermate wild type (WT) controls. The LFPs of the left and right secondary motor cortex (M2), specifically in the gamma band, were more synchronized in APP/PS1 mice than in WT controls, suggesting a declined hemispheric asymmetry of bilateral M2 in this AD mouse model. Notably, the recording and data analysis processes are flexible and easy to carry out, and can also be applied to other brain pathways when conducting experiments that focus on neuronal circuits.

We present in vivo electrophysiological recording of the local field potential (LFP) in bilateral secondary motor cortex (M2) of mice, which can be applied to evaluate hemisphere lateralization. The study revealed altered levels of synchronization between the left and right M2 in APP/PS1 mice compared to WT controls.

マウスの二次運動皮質における二国間局場電位記録による半球横化の評価

This article demonstrates complete, detailed procedures for both in vivo bilateral recording and analysis of local field potential (LFP) in the cortical ...

Investigating Long-term Synaptic Plasticity in Interlamellar Hippocampus CA1 by Electrophysiological Field Recording

The study of synaptic plasticity in the hippocampus has focused on the use of the CA3-CA1 lamellar network. Less attention has been given to the longitudinal interlamellar CA1-CA1 network. Recently however, an associational connection between CA1-CA1 pyramidal neurons has been shown. Therefore, there is the need to investigate whether the longitudinal interlamellar CA1-CA1 network of the hippocampus supports synaptic plasticity.
We designed a protocol to investigate the presence or absence of long-term synaptic plasticity in the interlamellar hippocampal CA1 network using electrophysiological field recordings both in vivo and in vitro. For in vivo extracellular field recordings, the recording and stimulation electrodes were placed in a septal-temporal axis of the dorsal hippocampus at a longitudinal angle, to evoke field excitatory postsynaptic potentials. For in vitro extracellular field recordings, hippocampal longitudinal slices were cut parallel to the septal-temporal plane. Recording and stimulation electrodes were placed in the stratum oriens (S.O) and the stratum radiatum (S.R) of the hippocampus along the longitudinal axis. This enabled us to investigate the directional and layer specificity of evoked excitatory postsynaptic potentials. Already established protocols were used to induce long-term potentiation (LTP) and long-term depression (LTD) both in vivo and in vitro. Our results demonstrated that the longitudinal interlamellar CA1 network supports N-methyl-D-aspartate (NMDA) receptor-dependent long-term potentiation (LTP) with no directional or layer specificity. The interlamellar network, however, in contrast to the transverse lamellar network, did not present with any significant long-term depression (LTD).

We used recording and stimulation electrodes in longitudinal hippocampal brain slices and longitudinally positioned recording and stimulation electrodes in the dorsal hippocampus in vivo to evoke extracellular postsynaptic potentials and demonstrate long-term synaptic plasticity along the longitudinal interlamellar CA1.

共局在脳波および齧歯動物のローカル フィールド電位の同時記録

共局在脳波および齧歯動物のローカルフィールド電位の同時記録