Name: recording electrical activity from identified neurons in the intact brain of transgenic fish
Uploaded: 2013-04-30T13:00:00
Description: Watch this Scientific Journal Video about Recording Electrical Activity from Identified Neurons in the Intact Brain of Transgenic Fish at JoVE.com

我々は技術支援のために博士孟チン林さんと元ドン感謝。この作品は、健康HD053767の国立研究所からの助成金（NLWに下請け）でサポートされている、研究担当副学長、カリフォルニア大学ロサンゼルス校（NLW）の生理学とOfficeの部門からの資金によってされました。

1。大人のメダカから脳の解剖<ol><li>に浸漬することにより（ 図1A）、男性または女性成人を麻酔5ミリリットルMS-222（150 mg / Lで、pHは7.4）、エラの動きがdecapitating前に中止した後、数分待ってください。すべての手順は、カリフォルニア大学ロサンゼルス校の制度動物実験委員会によって承認された。 </li><li> 60 mmの直径のペトリ皿にハサミで蓋の尾側端で魚の生理食塩水で魚の...

<ol>
	<li>Kah, O., Lethimonier, C., Lareyre, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gonadotrophin-releasing+hormone+(GnRH)+in+the+animal+kingdom.">Gonadotrophin-releasing hormone (GnRH) in the animal kingdom.</a> J. Soc. Biol. 198 (1), 53-60 (2004).</li><li>Ramakrishnan, S., Wayne, N. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Social+cues+from+conspecifics+alter+electrical+activity+of+gonadotropin-releasing+hormone+neurons+in+the+terminal+nerve+via+visual+signals.">Social cues from conspecifics alter electrical activity of gonadotropin-releasing hormone neurons in the terminal nerve via visual signals.</a> Am. J. Physiol. Regul. Integr. Comp. Physiol. 297 (1), R135-R141 (2009).</li><li>Abe, H., Oka, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+of+neuromodulation+by+a+nonhypophysiotropic+GnRH+system+controlling+motivation+of+reproductive+behavior+in+the+teleost.">Mechanisms of neuromodulation by a nonhypophysiotropic GnRH system controlling motivation of reproductive behavior in the teleost.</a> 57 (6), 665-674 (2011).</li><li>Oka, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tetrodotoxin-resistant+persistent+Na++current+underlying+pacemaker+potentials+of+fish+gonadotrophin-releasing+hormone+neurones.">Tetrodotoxin-resistant persistent Na+ current underlying pacemaker potentials of fish gonadotrophin-releasing hormone neurones.</a> J. Physiol. 482 (Pt. 1), 1-6 (1995).</li><li>Zhao, Y., Wayne, N. 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L., Kuwahara, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Beta-endorphin+alters+electrical+activity+of+gonadotropin+releasing+hormone+neurons+located+in+the+terminal+nerve+of+the+teleost+medaka+(Oryzias+latipes.">Beta-endorphin alters electrical activity of gonadotropin releasing hormone neurons located in the terminal nerve of the teleost medaka (Oryzias latipes.</a> Gen. Comp. Endocrinol. 150 (1), 41-47 (2007).</li><li>Oka, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three+types+of+gonadotrophin-releasing+hormone+neurones+and+steroid-sensitive+sexually+dimorphic+kisspeptin+neurones+in+teleosts.">Three types of gonadotrophin-releasing hormone neurones and steroid-sensitive sexually dimorphic kisspeptin neurones in teleosts.</a> J. 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GnRHの3：GFPトランスジェニック魚は、直接·間接的に再生3、8-10に関与している行動の中央制御の神経統合や規制の基礎となる神経生理学的メカニズムを研究するためのユニークなモデルを提供します。このモデルシステムの重要な利点の一つは、12、神経回路6を中断することなく、電気生理学的記録のための神経細胞、9、11には比較的簡単にアクセス?...

<table border="1">
 <tbody>
 <tr>
 <td>Name</td>
 <td>Company</td>
 <td>Catalog Number</td>
 <td>Comments</td>
 </tr>
 <tr>
 <td>Microscope</td>
 <td>Olympus</td>
 <td></td>
 <td>BX50W (Upright)</td>
 </tr>
 <tr>
 <td>Amplifier</td>
 <td>Axon Instruments</td>
 <td></td>
 <td>Axoclamp 200B </td>
 </tr>
 <tr>
 <td>A-D converter</td>
 <td>Computer Interference Corp.</td>
 <td></td>
 <td>Digidata ITC-18 </td>
 </tr>
 <tr>
 <td>Cooled CCD camera</td>
 <td>PCO Computer Optics </td>
 <td></td>
 <td>Sensicam</td>
 </tr>
 <tr>
 <td>Xenon lamp</td>
 <td>Sutter Instruments Co.</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>GFP filter set</td>
 <td>Chroma Technologies</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Imaging Software</td>
 <td>Intelligent Imaging Innovations</td>
 <td></td>
 <td>Slidebook software </td>
 </tr>
 <tr>
 <td>Electrophysiology Data Acquisition Software</td>
 <td>Axon Instruments</td>
 <td></td>
 <td>Axograph software </td>
 </tr>
 <tr>
 <td>Electrophysiology Data Acquisition Software</td>
 <td>AD Instruments Inc.</td>
 <td></td>
 <td>PowerLab </td>
 </tr>
 <tr>
 <td>Headstage for electrophysiology</td>
 <td>Axon Instruments</td>
 <td></td>
 <td>CV 203BU </td>
 </tr>
 <tr>
 <td>Micromanipulator</td>
 <td>Sutter Instrument Co </td>
 <td>MP-285 </td>
 <td></td>
 </tr>
 <tr>
 <td>Recording Chamber Platform</td>
 <td>Warner Instrument Corp.</td>
 <td></td>
 <td>P1 </td>
 </tr>
 <tr>
 <td>Recording Chamber</td>
 <td>Warner Instrument Corp.</td>
 <td></td>
 <td>RC-26G </td>
 </tr>
 <tr>
 <td>Electrode Puller</td>
 <td>Sutter instruments</td>
 <td></td>
 <td>P87 </td>
 </tr>
 <tr>
 <td>Filament for electrode puller</td>
 <td>Sutter Instruments</td>
 <td>FB330B</td>
 <td>3.0 mm wide trough filament</td>
 </tr>
 <tr>
 <td>1.5 mm glass capillaries</td>
 <td>World Precision Instruments</td>
 <td>1B150-4</td>
 <td>Microelectrode for recording</td>
 </tr>
 <tr>
 <td>Syringe</td>
 <td>Becton Dickinson</td>
 <td>309586</td>
 <td>3 ml</td>
 </tr>
 <tr>
 <td>MS-222 </td>
 <td>Sigma</td>
 <td>E10521-10G</td>
 <td>Ethyl 3-aminobenzoate methanesulfonate salt</td>
 </tr>
 <tr>
 <td>Fish saline</td>
 <td></td>
 <td></td>
 <td>mM: 134 NaCl; 2.9 KCl; 2.1 CaCl2; 1.2 MgCl2; 10 HEPES </td>
 </tr>
 <tr>
 <td>Electrode solution (loose-patch) </td>
 <td></td>
 <td></td>
 <td>mM: 150 NaCl; 3.5 KCl; 2.5 CaCl2; 1.3 MgCl2; 10 HEPES; 10 glucose </td>
 </tr>
 <tr>
 <td>Electrode solution (whole-cell patch) </td>
 <td></td>
 <td></td>
 <td>mM: 112.5 K-gluconate; NaCl; 17.5 KCl; 0.5 CaCl2; 1 MgCl2; 5 MgATP; 1 EGTA; 10 HEPES; 1 GTP; 0.1 leupeptin;10 phospho-creatine </td>
 </tr>
 </tbody>
</table>

recording electrical activity from identified neurons in the intact brain of transgenic fish

複雑な行動を調節する神経回路の細胞生理学を理解することは、この作業はCNSの神経回路は無傷のままで無傷の脳の調製において実行可能なモデルシステムを使用して、大幅に高められる。私たちは、ゴナドトロピン放出ホルモン（GnRH）ニューロンは遺伝的に無傷の脳内の識別のための緑色蛍光タンパク質でタグ付けされたトランスジェニック魚を使用しています。魚のGnRHニューロンの複数の集団があり、その機能は、脳内の場所と、彼らは1を表現することのGnRH遺伝子に依存しています。我々は、トランスジェニックメダカの無傷の脳（ 図1BおよびC）を使用して、嗅球に関連付けられているターミナル·神経（TN）に位置GnRH3ニューロンに私たちのデモを当てている。研究はメダカTN-GnRH3ニューロンは外部環境から中枢神経系への情報の送信器として作用する、神経調節であることを示唆している。トンちょっとなどのよく知られた視床下部GnRH1ニューロン2、3を実行します。TN-GnRH3ニューロンの自発活動電位の発火のトニックパターンが固有の特性4-6、周波数であり、下垂体-性腺機能の調節に直接的な役割を果たしていない同種2と神経ペプチドキスペプチン1 5からの視覚的な手がかりによって変調となっている。このビデオでは、TN-GnRH3のニューロンは神経細胞を識別し、脳全体での電気的活動を監視する方法をお見せするために強化された緑色蛍光タンパク質7にリンクGnrh3のプロモーター領域を含む導入遺伝子を発現するトランスジェニックメダカの安定したラインを使用準備6。

メダカの摘出脳からGFP標識TN-GnRH3ニューロンの二国間のクラスタの例を図1B及び1Cに示されている。各クラスタは、約8-10のGnRHニューロンを含んでいます。ターゲットTN-GnRH3の自発神経活動は、0.5〜6 Hzでの典型的な発火率と電流クランプモード（I = 0）で記録した。活動電位の発火のパターンはかなり規則的なスパイク間間隔で、通常はトニックや鼓動パターンです。?...

Watch this Scientific Journal Video about Recording Electrical Activity from Identified Neurons in the Intact Brain of Transgenic Fish at JoVE.com

Recording Electrical Activity from Identified Neurons in the Intact Brain of Transgenic Fish (Video) | JoVE

このビデオでは、複雑な神経回路を維持し、脳全体の準備、で識別される単一のニューロンからの電気的活動を記録する方法を説明します。私たちは、ゴナドトロピン放出ホルモン（GnRH）ニューロンは遺伝的に無傷の脳の準備の識別のための蛍光タンパク質でタグ付けされたトランスジェニック魚を使用しています。

トランスジェニックフィッシュの無傷の脳内で識別ニューロンからの電気的活動を記録する

Understanding the cell physiology of neural circuits that regulate complex behaviors is greatly enhanced by using model systems in which this work can be ...

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Neuroscience

Recording Electrical Activity from Identified Neurons in the Intact Brain of Transgenic Fish

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同定されたニューロンと摂食行動からカルシウム信号の同時記録キイロショウジョウバエ</em

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Intracellular Recording, Sensory Field Mapping, and Culturing Identified Neurons in the Leech, Hirudo medicinalis (Video) | JoVE

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 ...

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無傷アダルトゼブラフィッシュの脳における電気生理学的記録

Previously, electrophysiological studies in adult zebrafish have been limited to slice preparations or to eye cup preparations and electrorentinogram ...

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へのガイド<em&gt;インビボ</emOptogenetically同定された皮質抑制性介在から&gt;シングル単位記録

A major challenge in neurophysiology has been to characterize the response properties and function of the numerous inhibitory cell types in the cerebral ...

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無傷の神経上皮におけるマウス嗅細胞の穿孔パッチクランプ記録：同定された嗅覚受容体を発現するニューロンの機能解析

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の嗅球にカルシウム変化をモニタリングを通じて温度によって誘発される神経活動を記録<em&gt;アフリカツメガエル</em

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Imaging Neural Activity in the Primary Somatosensory Cortex Using Thy1-GCaMP6s Transgenic Mice

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