Name: external excitation of neurons using electric and magnetic fields in one- and two-dimensional cultures
Uploaded: 2017-05-07T14:00:00
Description: Watch this Scientific Journal Video about External Excitation of Neurons Using Electric and Magnetic Fields in One- and Two-dimensional Cultures at JoVE.com

著者たちは、Ofer Feinerman、Fred Wolf、Menahem Segal、Andreas Neef、Eitan Reuvenyに非常に有益な議論に感謝します。著者は、この技術の初期バージョンの開発について、Ilan BreskinとJordi Sorianoに感謝します。作者は理論的概念の助けを借りてTsvi TlustyとJean-Pierre Eckmannに感謝します。この研究は、ミネルバ財団、イスラエル科学技術省、イスラエル科学財団助成金1320/09およびバイナショナル科学財団助成金2008331によって支援された。

倫理文：動物の扱いを含む手順は、ワイツマン科学研究所の所内動物管理使用委員会（IACUC）、および適切なイスラエルの法律のガイドラインに従って行われました。ワイツマン研究所は、実験動物管理・インターナショナルの評価と認定協会（AAALAC）の認定を受けています。ワイツマン制度動物実験委員会は、海馬神経細胞で行ったこの研究を、承認しました。 （2D）2?...

<ol>
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1Dパターニングは、様々な用途に使用することができる重要なツールです。例えば、我々は、ラット海馬ニューロン5のクロナキシーおよび基電流を測定するために、より最近ニューロン培養物29からの論理ゲートを作成するための1次元パターニングを使用している、とと比較してダウン症候群海馬ニューロンにおける発火活動の信号伝搬速度の減速します...

ニューロンおよび外部電場との相互作用は、基本的な意味合いだけでなく、実用的なものを持っています。それは外部から印加される電界が組織を励起することができるボルタの時代から知られているが、ニューロンにおける結果として得られる活動電位の生成を担うメカニズムは、ごく最近、4 3、2、1解明され始めています。これは、電場2、5に応答ニューロンにおける膜電位の脱分極、膜特性のとイオンチャネルの役割、さらには地域の原因となるメカニズムに関する質問への回答を見つけることが含まれます。治療的神経刺激6、7、8、9、 <supクラス=「外部参照」は&gt; 10個の方法論は、被災地を標的とすると治療の結果を理解するために重要であることができ、この情報に特に依存しています。このような理解はまた、治療プロトコルと、脳内の異なる領域を刺激するための新しいアプローチを開発するのに役立ちます。 in vivoでの脳内の相互作用を測定することは、この理解に重要な要素が追加されますが、頭蓋骨内の測定の不正確さと低制御性によって妨げられています。対照的に、培養物中の測定を容易に高精度、ノイズ性能に優れた信号と再現性と制御性の高い大量に行うことができます。集合的ネットワーク動作のニューロン特性の多種多様を14、13、12、11明らかにすることができる培養物を使用して、 </sup&gt; 15、16。同様に、このよく制御されたシステムは、他の刺激方法がoptogeneticallyアクティブニューロン17、18の光刺激の間のチャネル開口部、19は活動電位を生成するための責任がある方法、例えば、動作するメカニズムを解明するには非常に効率的であることが期待されます。 ここでの焦点は、効率的に外部電界を経由してニューロンを励起することができるツールの開発と理解を記述するにあります。本稿では、バス電極によって異なる構成と直接印加される電界の向きを使用して、刺激を二次元の調製と一次元パターン化された海馬培養物を記述し、そして最終的に二次元の刺激とによって一次元培養をパターニング時間的に変化する電界を誘起する磁界を、5、20、21。

<table>
	<tbody>
		<tr>
			<td>APV</td>
			<td>Sigma-Aldrich</td>
			<td>A8054</td>
			<td>Disconnect the network. Mentioned in Section 2.4.2</td>
		</tr>
		<tr>
			<td>B27 supp</td>
			<td>Gibco</td>
			<td>17504-044</td>
			<td>Plating medium. Mentioned in Section 1.1.1</td>
		</tr>
		<tr>
			<td>bicuculline</td>
			<td>Sigma-Aldrich</td>
			<td>14343</td>
			<td>Disconnect the network&#160;. Mentioned in Section 2.4.2</td>
		</tr>
		<tr>
			<td>Borax (sodium tetraborate decahydrate)</td>
			<td>Sigma-Aldrich</td>
			<td>S9640</td>
			<td>Borate buffer. Mentioned in Section 1.1.2</td>
		</tr>
		<tr>
			<td>Boric acid</td>
			<td>Frutarom LTD</td>
			<td>5550710</td>
			<td>Borate buffer. Mentioned in Section 1.1.2</td>
		</tr>
		<tr>
			<td>CaCl2 , 1M</td>
			<td>Fluka&#160;</td>
			<td>21098</td>
			<td>Extracellular recording solution&#160;. Mentioned in Section 1.5.2</td>
		</tr>
		<tr>
			<td>CNQX</td>
			<td>Sigma-Aldrich</td>
			<td>C239</td>
			<td>Disconnect the network&#160;. Mentioned in Section 2.4.2</td>
		</tr>
		<tr>
			<td>COMSOL</td>
			<td>COMSOL Inc</td>
			<td>Multiphysics 3.5</td>
			<td>Numerical simulation. Mentioned in Section 3.5.2</td>
		</tr>
		<tr>
			<td>D-(+)-Glucose, 1M</td>
			<td>Sigma-Aldrich</td>
			<td>65146</td>
			<td>Plating medium, Extracellular recording solution&#160;. Mentioned in Section 1.1.1&#160;&#160;&#160; 1.5.2</td>
		</tr>
		<tr>
			<td>D-PBS</td>
			<td>Sigma-Aldrich</td>
			<td>D8537</td>
			<td>Cell Cultures. Mentioned in Section 1.2.4&#160;&#160;&#160; 1.2.6</td>
		</tr>
		<tr>
			<td>FCS(FBS)</td>
			<td>Gibco</td>
			<td>12657-029</td>
			<td>Plating medium. Mentioned in Section 1.1.1</td>
		</tr>
		<tr>
			<td>Fibronectin</td>
			<td>Sigma-Aldrich</td>
			<td>F1141</td>
			<td>Bio Coating. Mentioned in Section 1.2.6</td>
		</tr>
		<tr>
			<td>Fluo4, AM</td>
			<td>Life technologies</td>
			<td>F14201</td>
			<td>Imaging of spontaneous or evoked activity&#160;. Mentioned in Section 1.5.1&#160;&#160;&#160; 1.5.3&#160;&#160;&#160; 1.5.5</td>
		</tr>
		<tr>
			<td>FUDR</td>
			<td>Sigma-Aldrich</td>
			<td>F0503</td>
			<td>Changing medium. Mentioned in Section 1.4.1</td>
		</tr>
		<tr>
			<td>Gentamycin</td>
			<td>Sigma-Aldrich</td>
			<td>G1272</td>
			<td>Plating medium, Changing medium, Final medium. Mentioned in Section 1.1.1</td>
		</tr>
		<tr>
			<td>GlutaMAX 100X</td>
			<td>Gibco</td>
			<td>35050-038</td>
			<td>Plating medium, Changing medium, Final medium. Mentioned in Section 1.1.1</td>
		</tr>
		<tr>
			<td>Hepes, 1M</td>
			<td>Sigma-Aldrich</td>
			<td>H0887</td>
			<td>Extracellular recording solution&#160;. Mentioned in Section 1.5.2</td>
		</tr>
		<tr>
			<td>HI HS&#160;</td>
			<td>BI</td>
			<td>04-124-1A</td>
			<td>Plating medium, Changing medium, Final medium. Mentioned in Section 1.1.1&#160;&#160;&#160; 1.4.1&#160;&#160;&#160; 1.4.2</td>
		</tr>
		<tr>
			<td>KCl,&#160; 3M</td>
			<td>Merck</td>
			<td>1049361000</td>
			<td>Extracellular recording solution. Mentioned in Section 1.5.2</td>
		</tr>
		<tr>
			<td>Laminin&#160;</td>
			<td>Sigma-Aldrich</td>
			<td>L2020</td>
			<td>Bio Coating. Mentioned in Section 1.2.6</td>
		</tr>
		<tr>
			<td>MEM x 1</td>
			<td>Gibco</td>
			<td>21090-022</td>
			<td>Plating medium, Changing medium, Final medium. Mentioned in Section 1.4.1&#160;&#160;&#160; 1.4.2</td>
		</tr>
		<tr>
			<td>MgCl2 , 1M</td>
			<td>Sigma-Aldrich</td>
			<td>M1028</td>
			<td>Extracellular recording solution. Mentioned in Section 1.5.2</td>
		</tr>
		<tr>
			<td>NaCl, 4M</td>
			<td>Bio-Lab</td>
			<td>19030591</td>
			<td>Extracellular recording solution&#160;. Mentioned in Section 1.5.2</td>
		</tr>
		<tr>
			<td>Octadecanethiol</td>
			<td>Sigma-Aldrich</td>
			<td>01858</td>
			<td>Cleaning Cr-Au coated coverslips (1D cultures). Mentioned in Section 1.2.3</td>
		</tr>
		<tr>
			<td>Pluracare F108 NF Prill</td>
			<td>BASF Corparation&#160;</td>
			<td>50475278</td>
			<td>Bio-Rejection Coating, Bio Coating. Mentioned in Section 1.2.4&#160;&#160;&#160; 1.2.6</td>
		</tr>
		<tr>
			<td>Poly-L-lysine 0.01% solution&#160;</td>
			<td>Sigma-Aldrich</td>
			<td>&#160;P47075</td>
			<td>Promote cell division. Mentioned in Section 1.1.4</td>
		</tr>
		<tr>
			<td>Sucrose, 1M</td>
			<td>Sigma-Aldrich</td>
			<td>S1888</td>
			<td>Extracellular recording solution&#160;. Mentioned in Section 1.5.2</td>
		</tr>
		<tr>
			<td>Thiol&#160;</td>
			<td>Sigma-Aldrich</td>
			<td>1858</td>
			<td>Bio-Rejection Coating. Mentioned in Section 1.2.3</td>
		</tr>
		<tr>
			<td>URIDINE</td>
			<td>Sigma-Aldrich</td>
			<td>U3750</td>
			<td>Changing medium. Mentioned in Section 1.4.1</td>
		</tr>
		<tr>
			<td>Sputtering machine</td>
			<td>AJA International, Inc</td>
			<td>ATC Orion-5Series&#160;</td>
			<td>coating glass with thin layers of metal. Mentioned in Section 1.2.2</td>
		</tr>
		<tr>
			<td>Pen plotter&#160;</td>
			<td>Hewlett Packard&#160;</td>
			<td>HP 7475A</td>
			<td>Etching of pattern to the coated coverslip. Mentioned in Section 1.2.5</td>
		</tr>
		<tr>
			<td>Electrodes wires&#160;</td>
			<td>A-M Systems, Carlsborg WA</td>
			<td>767000</td>
			<td>Electric stimulation of neuronal cultures. Mentioned in Section 2.1&#160;&#160;&#160; 2.2&#160;&#160;&#160; 2.3&#160;&#160; 2.4.5</td>
		</tr>
		<tr>
			<td>Signal generator</td>
			<td>BKPrecision</td>
			<td>4079</td>
			<td>Shaping of the electric signal. Mentioned in Section 2.3</td>
		</tr>
		<tr>
			<td>Amplifier</td>
			<td>Homemade</td>
			<td></td>
			<td>Voltage amplification of the signal from the signal generator to the electrodes. Mentioned in Section 2.3</td>
		</tr>
		<tr>
			<td>Power supply</td>
			<td>Matrix&#160;</td>
			<td>MPS-3005 LK-3&#160;</td>
			<td>Power supply to the sputtering machine. Mentioned in Section 1.2.2.3</td>
		</tr>
		<tr>
			<td>Transcranial magnetic stimulation</td>
			<td>Magstim, Spring Gardens, UK</td>
			<td>Rapid 2</td>
			<td>Magnetic stimulation of neuronal culture. Mentioned in Section 3.1&#160;&#160; 3.3&#160;&#160; 3.4</td>
		</tr>
		<tr>
			<td>Epoxy</td>
			<td>Cognis</td>
			<td>Versamid 140</td>
			<td>Casting of homemade coils. Mentioned in Section 3.4</td>
		</tr>
		<tr>
			<td>Epoxy</td>
			<td>Shell</td>
			<td>EPON 815&#160;</td>
			<td>Casting of homemade coils. Mentioned in Section 3.4</td>
		</tr>
		<tr>
			<td>Platinum wires 0.005&#39;&#39; thick; A-M Systems,&#160;</td>
			<td>&#160;Carlsborg WA</td>
			<td>&#160;767000</td>
			<td>Electric stimulation of neuronal cultures. Mentioned in Section 2.1</td>
		</tr>
		<tr>
			<td>Circular magnetic coil</td>
			<td>Homemade</td>
			<td></td>
			<td>Magnetic stimulation of neuronal culture. Mentioned in Section 3.3</td>
		</tr>
		<tr>
			<td>WaveXpress SW</td>
			<td>B&#38;K Precision&#160;</td>
			<td></td>
			<td>Waveform editing software. Mentioned in Section 2.1.32</td>
		</tr>
		<tr>
			<td>Xion Ultra 897</td>
			<td>Andor</td>
			<td></td>
			<td>Sensitive EMCCD camera. Mentioned in Section 2.4.4</td>
		</tr>
	</tbody>
</table>

external excitation of neurons using electric and magnetic fields in one- and two-dimensional cultures

その膜電位が一定の閾値を超えた場合、ニューロンは活動電位を発射します。脳の典型的な活動では、これは、シナプスへの化学入力の結果として生じます。しかし、ニューロンも課せられ、電界によって励起することができます。特に、最近の臨床応用には、外部から電場を作成することによって、神経細胞を活性化させます。これは、ニューロンが外部磁場に応答して、何が活動電位が発生する方法を検討することが重要です。幸いなことに、外部電場の正確かつ制御されたアプリケーションは、切除解離、および培養物中で増殖させ、胚性神経細胞のために可能です。これは、再現性の高いシステムではこれらの質問の調査を可能にします。 本論文では、ニューロン培養物に対する外部電界の制御された適用のために使用される技術のいくつかが検討されています。ネットワークは、いずれか1次元、 すなわちリネアにパターニングすることができますR形態または基板の全体面上に成長させ、したがって、二次元。また、励起は、流体中に浸漬電極を介して電界の直接適用（バス電極）または磁気パルスのリモート作成を使用して電場を誘導することによって作成することができます。

<html>提示プロトコルは、神経細胞培養の容易なパターニングが可能になります。それは私たちが刺激のために開発され、いくつかの方法と組み合わせると、それは、健康と病気のニューロン27の特性を比較するための関数としての文化を刺激するための最適な方法を見つけるために、そのような時値および基電流5のように、いくつ?...

Watch this Scientific Journal Video about External Excitation of Neurons Using Electric and Magnetic Fields in One- and Two-dimensional Cultures at JoVE.com

External Excitation of Neurons Using Electric and Magnetic Fields in One- and Two-dimensional Cultures

神経細胞培養は、単一ニューロンまたはニューロンの人口への影響を通じて新興脳刺激技術を研究するための良いモデルです。ここに提示バス電極によって直接生成または時間変化する磁場によって誘起される電界によりパターニング神経細胞培養物の刺激のための異なる方法があります。

電気を使用してニューロンと1次元と2次元培養における磁場の外部励起

A neuron will fire an action potential when its membrane potential exceeds a certain threshold. In typical activity of the brain, this occurs as a result ...

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Dissection and Culture of Commissural Neurons from Embryonic Spinal Cord

胚の脊髄から交連ニューロンの解剖と文化

Commissural neurons have been widely used to investigate the mechanisms underlying axon guidance during embryonic spinal cord development. The cell bodies ...

Using Affordable LED Arrays for Photo-Stimulation of Neurons

ニューロンの光刺激のために手ごろな価格のLEDアレイを使用する

Standard slice electrophysiology has allowed researchers to probe individual components of neural circuitry by recording electrical responses of single ...

Comprehensive Profiling of Dopamine Regulation in Substantia Nigra and Ventral Tegmental Area

黒質や腹側被蓋野のドーパミン規制の包括的なプロファイリング

Dopamine is a vigorously studied neurotransmitter in the CNS. Indeed, its involvement in locomotor activity and reward-related behaviour has fostered five ...

One Mouse, Two Cultures: Isolation and Culture of Adult Neural Stem Cells from the Two Neurogenic Zones of Individual Mice

マウスを一回、二つの文化：個々のマウスの二つの神経原性のゾーンから成体神経幹細胞の単離および培養

The neurosphere assay and the adherent monolayer culture system are valuable tools to determine the potential (proliferation or differentiation) of adult ...

The Use of Magnetic Resonance Spectroscopy as a Tool for the Measurement of Bi-hemispheric Transcranial Electric Stimulation Effects on Primary Motor Cortex Metabolism

一次運動野代謝に及ぼすバイ半球経頭蓋電気刺激効果の測定のためのツールと​​しての磁気共鳴分光法の利用

Transcranial direct current stimulation (tDCS) is a neuromodulation technique that has been increasingly used over the past decade in the treatment of ...

Rapid Genotyping of Animals Followed by Establishing Primary Cultures of Brain Neurons

脳神経細胞の初代培養の確立が続く動物の急速なジェノタイピング

High-resolution analysis of the morphology and function of mammalian neurons often requires the genotyping of individual animals followed by the analysis ...

Reliable Identification of Living Dopaminergic Neurons in Midbrain Cultures Using RNA Sequencing and TH-promoter-driven eGFP Expression

RNA配列決定およびTH-プロモーター駆動EGFP発現を使用して中脳培養における生活ドーパミン作動性ニューロンの信頼性の高い同定

In Parkinson&#39;s Disease (PD) there is widespread neuronal loss throughout the brain with pronounced degeneration of dopaminergic neurons in the SNc, ...

Quantification of Endosome and Lysosome Motilities in Cultured Neurons Using Fluorescent Probes

蛍光プローブを用いた培養ニューロンにおけるエンドソームおよびリソソームの動力学の定量

In the brain, membrane trafficking systems play important roles in regulating neuronal functions, such as neuronal morphology, synaptic plasticity, ...

Preparations and Protocols for Whole Cell Patch Clamp Recording of Xenopus laevis Tectal Neurons

Preparations and Protocols for Whole Cell Patch Clamp Recording of Xenopus laevis Tectal Neurons

準備やアフリカツメガエルの視蓋ニューロンの全体セル パッチ ・ クランプ記録のためのプロトコル

The Xenopus tadpole retinotectal circuit, comprised of the retinal ganglion cells (RGCs) in the eye which form synapses directly onto neurons in the optic ...

Ballistic Labeling of Pyramidal Neurons in Brain Slices and in Primary Cell Culture

脳スライスと一次細胞培養における錐体ニューロンの弾道標識

It has been reported that the size and shape of dendritic spines is related to their structural plasticity. To identify the morphological structure of ...