著者は、Hyewonクーパーの芸術的な貢献に感謝しています。

<ol>
	<li>Whitear, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chordotonal+organs+in+Crustacea.">Chordotonal organs in Crustacea.</a> Nature. 187, 522-523 (1038).</li><li>Alexandrowicz, J. 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+comparative+anatomy+of+leg+propriocetors+in+some+decapod+Crustacea.">The comparative anatomy of leg propriocetors in some decapod Crustacea.</a> J. Mar. Biol. Assoc. U.K. 52 (3), 605-634 (1972).</li><li>Hartman, H. B., Boettiger, E. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+functional+organization+of+the+propys-dactylus+organ+in+Cancer+irroratus+Say.">The functional organization of the propys-dactylus organ in Cancer irroratus Say.</a> Comp. Biochem. 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著者は、彼らが競合する経済的利益を持っていないことを宣言します。

この一連の実験の目的は、1）特定の感覚系の生理的機能に関連する解剖学的マッピングの重要性を強調するために）識別可能な固有感覚器官の緊張神経と2から細胞外記録の基本原則を教え、示すことである。この実験的なアプローチを利用し、動物モデルは、安価で神経生理学教示実験室で行うことが比較的容易である。 弦音器官のニューロンは2特定の機能タイプ、動きに反応しているものと静的な位置に対応したものである。弦音器官の様々からの単一細胞記録は、関節が検討されているに関係なく、これが事実で3,5であることが示されている。実際、ロブスターの触角の関節に関連した弦音器官が同じ2つの感覚の種類と基本的な解剖学27を明らかにした。そこに二種類のニューロン（運動及びPOSITIであることに加えて）に、神経細胞は、それぞれの弾性ストランドの同じ解剖学的構造を共有しています。鎖上に近位にある大きな細胞体は、ダイナミックな動きに敏感な神経細胞に所属する傾向がある。静的な位置を知らせるニューロンは、小さな細胞体を持ち、遠位に配置されています。これらの細胞は持続的に活躍しています。手根-propodus（CP）と長節-手根（MC）関節の2弦音器官が存在する間は、PDジョイントは、単一の弦音の臓器が含まれています。 電気生理学的記録のためのワタリガニ（ クロストリジウム·サピドゥス ）に固有受容構造を公開する解剖は、感覚ニューロンから記録しながら、関節の動きが自然な位置で行われることを可能にする戦略が必要です。歩行脚での開幕戦の筋肉のためのテンション神経は、いくつかの神経細胞で構成された非常に細かい神経である。注意を取られていない限り、テンション神経だけでなく、運動神経が刺激される筋肉を支配する、この解剖中に破損することができます。のために最適な記録は、吸引電極が神経のサイズに合わせて調整する必要がある。録音は30-40X倍率解剖顕微鏡およびローエンドのマイクロマニピュレーターを使用して、学生の研究室で容易にアクセスできます。 共同弦音器官を追求するために興味深いものになるだろう将来の実験は、ライフサイクルのさまざまな段階で様々な種での脚再生中の構造や生理的なプロフィールを調べることであろうがんマジ 19,26を使用した初期の研究のフォローアップ。対処すべき残りの質問は、1アール）手足を再生する際、再生さニューロンの分布と組織は2）機能的な再生四肢におけるCNS（腹側神経索）に軸索投射されているか、またはそれをしない、動物の年齢に依存しない手足がautotときに機能的な接続を確立するのにかかった時間との共同使用を取り、3）自切面の近位に切断軸索がどうなるかomized？28 甲殻類は、環境条件およびその周囲の温度に適合するが、それはニューロンは、温度変化に応答してその活性を変化させるように、それらが神経回路内の協調を維持するかは不明である。急激な変化は、30をnot29あり、一方、変化の遅い速度は、動物に順応するためのいくつかの時間を与えることがあります。原因代謝、行動31、あるいは環境への影響へのpHや浸透圧の生理的変化は、固有感覚に関わる神経回路に同様の課題を提示することができる。その機能は、単一細胞レベルで十分に特徴付けられているので、これらの甲殻類の製剤は、この種の問題に対処するための理想的です。 このプロトコルでは、オープナーの筋肉によって生成される力を監視する張力ニューロンの生理学的重要性が実証されている。これらの張力受容体は、染色手順を用いて、内突起内のそれらの位置を追跡することができる。これらのニューロンは、哺乳動物におけるように、様々なレベルでの力を検出する力が増加するにつれて、追加の神経細胞を補充する。受信飽和に達するまで活性の周波数は、運動ニューロンの刺激周波数に関連する。曲げられ指節関節とクイックリリース·プロトコルを使用して、テンション活動は急速に消えますが、その後、完全に拡張された、関節に張りを取り戻す際に返します。これはテンション受容体によって測定された力を説明するための古典的な実験手順である。種々の神経修飾物質は、それが力および神経応答の発生に影響するかを見るために製剤に適用することができる。重要な側面の1つは、神経応答が処理され、中枢神経系および運動ニューロンの活性に及ぼす影響に集積されている方法である。私達が示されている技術には1が神経節とバックにそのまま脚の緊張（感覚）神経運動神経回路機能、 すなわち 、信号の詳細については、対処するために開始できるように筋肉へ。 実証染色手順は、固有受容器官を支配する感覚ニューロンの生理学を理解する鍵である。機能と相馬のサイズに基づいて、ニューロンの解剖学的配置は、カニの足の中の様々な弦音器官で類似している。同様のニューロンの配置は、他の甲殻類や昆虫の種に見出されるかどうかは知られていない。単一細胞からの生理的な録音を組み合わせ、場所をマッピングすることは、直接、構造と機能の関係を可能にします。のCoCl 2染色および固定した解剖学的構造の長期保存には1を繰り返し対策を行い、構造配置を評価することができます。 固有感覚と骨格筋の緊張の受信が骨格筋構成のさまざまな関節動物のための協調行動や、外部および内部環境への対応を可能にする感覚モダリティであるS。 ;ザリガニの腹部の筋肉の受容器官は、別の十分に文書化の準備（Crawdadプロジェクトが表示され<a href="http://www.crawdad.cornell.edu/">http://www.crawdad.cornell.edu/</a>腹部半セグメント23ごとに2つのニューロンと固有感覚の教育目的のために） 。感覚神経束への単一ニューロンから記録することができることは、感覚受信の基本原理の理解を助けることが更なる詳細を提供します。これらの比較的単純な甲殻類の製剤は、1が自己受容や他の感覚入力9月12日、32、33の中央統合を可能にする神経回路を決定する可能性のある、固有感覚と緊張の監視の基本的な側面に対処することができます。

固有感覚は、協調運動行動を可能に四肢の位置や動きの感覚である。固有受容位置（静的）と動き（運動感覚）の受容体で構成されています。昆虫や甲殻類では、弦音器官は、CNS 1にその情報を提供する構造体である。すべての弦音器官は、関節にまたがるが、彼らはまだ、関節にまたがると、骨格筋や関節の関節に連動して移動するapodemes（構造のような腱）上のそれらの結合のために、関節の動きを監視することができます。カニの脚部は、それぞれが1つまたは2つの弦音器官2を有する、6個の関節を有している。通常、弦音器官は動き3-6の弾性ストランド、静的な関節位置を知らせるニューロン、方向と速度内に埋め込 ​​ま60〜100以上の感覚神経を持っています。各関節と脚の弦音器官からの入力は、その後、集中的に動物による協調の動きを可能に処理される。 <p class="jove_content"&gt;足の筋肉が等尺性と等張性収縮時の生産力は、筋線維とapodemes7-9にその添付ファイルに関連付けられたテンション受容体によって検出される。我々は固有感覚を監視一次感覚ニューロンからの記録と筋線維によって発生する力に応答するニューロンのための方法論を提示従っ甲殻類の歩行脚プロトコルでは。足の動きを活性化し、力発生を定量化するための技術も提示、ならびにこれらの末梢神経系構造体の配置を特徴付けるために使用され得る、解剖学的技法れる。 以下に示す手順では、弦音弾性ストランドと内突起上の位置を基準に両タイプの受容体を刺激するニューロンの構造的および機能解析を可能にします。説明するために、我々はpropodite -指節（PD）弦音臓器、カニ脚3の最も遠位のセグメントにまたがる臓器を使用しています。詳細な電気生理学的研究は1930年に始まり、現在も行われているものの、いくつかの側面は、様々な関節とmuscles10-16の協調制御における役割における固有受容の分節の接続については知られるようになった。固有受容器官、筋肉や神経系の間の構造と機能の関係を確立することは、さらに、これらの役割を定義するのに役立ちます。例えば、内突起に挿入細胞体とテンションニューロンの遠位終末を標識化することは、筋繊維8,17-21との相対的な位置を明らかにします。 我々は、学術研究や実験室で使用することができる脚甲殻類のための3つの染色技術を提示する。メチレンブルー染色は、筋肉や神経に適しコントラストを提供し、解剖学を学ぶ学生のための簡単​​な手法として推奨されます。蛍光顕微鏡のセットアップを持っているラボは簡単に神経を露出させることにより、より選択的な神経細胞の染色を達成することができます生体色素4 - ジ-2-ASPにS。第三の選択肢は、ニューロンを染色すると修正のCoCl 2埋め戻し、であり、蛍光イメージングを必要としません。それは労力と時間集約的でありながら、この染色プロセスが満たされている神経のための高コントラストと特異性を提供します。一緒に、これらの技術は、四肢または四肢内の間だけでなく、他の甲殻類や昆虫種20〜22のうち、種々の弦音器官を比較するために使用することができる。生理的なレコーディングにと解剖学的染色に用いワタリガニ（C.のサピドゥス ） は、すべての米国の南部と南東境界付近容易に入手可能である。この種は、ほとんどのカニで発見弦音と緊張神経手配の代表として機能します。西海岸の研究所では、これらの実験のためにはるかに大きいアメリカイチョウガニ（ 蟹座マジスター ） を使用することを好むでしょう。

<table border="0" cellpadding="0" cellspacing="0" width="686">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Sylgard</td>
			<td style="width:184px;">Dow Corning</td>
			<td style="width:119px;">182 silicone kit</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">NaCl</td>
			<td style="width:184px;">Sigma-Aldrich</td>
			<td style="width:119px;">S7653</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">KCl</td>
			<td style="width:184px;">Sigma-Aldrich</td>
			<td style="width:119px;">P9333</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">CaCl2</td>
			<td style="width:184px;">Sigma-Aldrich</td>
			<td style="width:119px;">C5670</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">HEPES acid</td>
			<td style="width:184px;">Sigma</td>
			<td style="width:119px;">3375</td>
			<td style="width:167px;">Free acid, crystalline</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">HEPES base</td>
			<td style="width:184px;">Sigma</td>
			<td style="width:119px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">D-Glucose</td>
			<td style="width:184px;">Sigma</td>
			<td style="width:119px;">G7021</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:216px;">MgSO4&#8226;7H2O</td>
			<td style="width:184px;">Sigma</td>
			<td style="width:119px;">M2643</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:216px;">Na2SO4&#8226;10H2O</td>
			<td style="width:184px;">Sigma</td>
			<td style="width:119px;">246980</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Bouin&#8217;s solution fixative</td>
			<td style="width:184px;">Sigma</td>
			<td style="width:119px;">HT10-1-32</td>
			<td style="width:167px;">Caution: Hazardous material (Special shipping cost required)</td>
		</tr>
		<tr height="105">
			<td height="105" style="height:105px;width:216px;">CoCl2</td>
			<td style="width:184px;">Sigma</td>
			<td style="width:119px;"></td>
			<td style="width:167px;">Caution: Hazardous material. Please follow proper disposal according to local and federal regulations.</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Methylene blue chloride</td>
			<td style="width:184px;">Matheson Co., Inc</td>
			<td style="width:119px;">Basic Blue 9, C.I. 52015</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">4-Di-2-ASP</td>
			<td style="width:184px;">Molecular Probes</td>
			<td style="width:119px;"></td>
			<td style="width:167px;">4-(4-diethylaminostyryl)-N-methylpyridinium iodide</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Bleach</td>
			<td style="width:184px;">Sigma-Aldrich</td>
			<td style="width:119px;"></td>
			<td style="width:167px;">To chloride silver wire</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">NaOH</td>
			<td style="width:184px;">Sigma-Aldrich</td>
			<td style="width:119px;">221465</td>
			<td style="width:167px;">To adjust pH</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">HCl</td>
			<td style="width:184px;">Sigma-Aldrich</td>
			<td style="width:119px;">H1758</td>
			<td style="width:167px;">To adjust pH</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;"></td>
			<td style="width:184px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Materials</td>
			<td style="width:184px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Dissecting tools</td>
			<td style="width:184px;">World Precision Instruments</td>
			<td style="width:119px;">assortment</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Intracellular electrode probe</td>
			<td style="width:184px;"></td>
			<td style="width:119px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Faraday cage</td>
			<td style="width:184px;"></td>
			<td style="width:119px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Insect Pins</td>
			<td style="width:184px;">Fine Science Tools, Inc</td>
			<td style="width:119px;">26001-60</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Dissecting microscope (100X)</td>
			<td style="width:184px;"></td>
			<td style="width:119px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Fiber optic lamp</td>
			<td style="width:184px;"></td>
			<td style="width:119px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Small adjustable mirror</td>
			<td style="width:184px;"></td>
			<td style="width:119px;"></td>
			<td style="width:167px;">To direct light beam toward the preparation.</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Glass electrodes</td>
			<td style="width:184px;">Sigma-Aldrich</td>
			<td style="width:119px;">CLS7095B5X</td>
			<td style="width:167px;">Box of 200, suction electrodes</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Micromanipulator</td>
			<td style="width:184px;">World Precision Instruments</td>
			<td style="width:119px;">MD4-M3-R</td>
			<td style="width:167px;">Can fix for base or on a metal rod</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Raised preparation stand</td>
			<td style="width:184px;"></td>
			<td style="width:119px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Silver wire (10/1,000 inch)</td>
			<td style="width:184px;">A-M Systems</td>
			<td style="width:119px;">782500</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Computer</td>
			<td style="width:184px;">Any company</td>
			<td style="width:119px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">AC/DC differential amplifier</td>
			<td style="width:184px;">A-M Systems</td>
			<td style="width:119px;">Model 3000</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">PowerLab 26T</td>
			<td style="width:184px;">AD Instruments</td>
			<td style="width:119px;">27T</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Force transducer</td>
			<td style="width:184px;">AD Instruments</td>
			<td style="width:119px;">0-50g</td>
			<td style="width:167px;">MLTF050/ST</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Head stage</td>
			<td style="width:184px;">AD Instruments</td>
			<td style="width:119px;"></td>
			<td style="width:167px;">Comes with AC/DC amplifier</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">LabChart7</td>
			<td style="width:184px;">AD Instruments</td>
			<td style="width:119px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Electrical leads</td>
			<td style="width:184px;">Any company</td>
			<td style="width:119px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Glass tools</td>
			<td style="width:184px;">Make yourself</td>
			<td style="width:119px;"></td>
			<td style="width:167px;">For manipulating nerves</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Cable and connectors</td>
			<td style="width:184px;">Any company</td>
			<td style="width:119px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Pipettes with bulbs</td>
			<td style="width:184px;">Fisher Scientific</td>
			<td style="width:119px;">13-711-7</td>
			<td style="width:167px;">Box of 500</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Beakers</td>
			<td style="width:184px;">Any company</td>
			<td style="width:119px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Wax or modeling clay</td>
			<td style="width:184px;">Any company or local stores</td>
			<td style="width:119px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Stimulator</td>
			<td style="width:184px;">Grass Instruments</td>
			<td style="width:119px;">SD9 or S88</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="42">
			<td height="84" rowspan="2" style="height:84px;width:216px;">Plastic tip for suction electrode</td>
			<td rowspan="2" style="width:184px;">Local hardware store (Watt&#8217;s brand)</td>
			<td rowspan="2" style="width:119px;">&#188; inch OD x 0.170 inch ID</td>
			<td style="width:167px;">Cut in small pieces. Pull out over a flame and cut back the tip to the correct size</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:167px;"></td>
		</tr>
	</tbody>
</table>

proprioception and tension receptors in crab limbs: student laboratory exercises

これらの手順の主な目的は、彼らは関節位置や動きを検出し、筋肉の緊張しているように固有感覚を担当する一次感覚ニューロンを生きているの活動を記録する方法を教育や研究目的のために実証することである。甲殻類固有受容テンション受容体から電気的活動は、基本的な神経生理学計装によって記録され、トランスデューサは、同時に運動神経を刺激することによって生成される力を測定するために使用される。また、我々は彼らの解剖学的構造の迅速な評価のために、または恒久的な固定のために神経細胞を染色する方法を示します。染色は、ほとんどの甲殻類での弦音器官の代表である解剖学的組織を明らかにする。固有受容応答にテンション神経の応答を比較すると、これらの感覚ニューロンが機能的に定義される方法と解剖学が関数に相関しているか決定する上で効果的な教育ツールである。三つの染色技術研究者やインストラクターが自分の研究室のための理想的な方法を選択することができています。

PD器官が完全に関節を拡張することにより延伸すると、図10における第1秒間に示すように、PDの神経における活動は、移動中に頑強である。それが開いた位置にまだ保持されている間、いくつかの活性が残っている。この活動は、静止位置感受性ニューロン（ 図10に示した記録の後半）からのものである。運動は、移動中の応答を呼び起こす、焼成は弦音鎖（ 図11）を延伸する際、主に存在している。 スパイクのさらなる分析が容易に相対的な振幅をソートすることにより近づけることができる。これは運動5の位置や種類のために募集された感覚ニューロンの異なる集団を実証するためのアプローチです。典型的な振幅は0.25〜1.5 mVの範囲が、これらの値は、吸引電極シールの抵抗（ すなわち気密性 ​​）に依存している。 T内のスパイクの周波数彼は様々な大きさの範囲はまた、分析のためにグラフィカルに表すことができる。 刺激周波数に対するオープナーの筋肉によって生成された力は、（ 図13AおよびB）を互いの上にそれぞれの電圧-時間トレースを重ね合わせて比較することができる。これはまた、それぞれの所与の刺激周波数の各関節位置のために行うことができる。テンション神経の活性を、各刺激周波数で生成された相対力の量と相関しており、各関節位置のためにすることができる。 PDの神経のように、スパイクの振幅の様々なオープナー筋（ 図13C）の収縮に応答して見られる。 歩行脚におけるニューロンの解剖学的配置を明確にメチレンブルー染色（ 図14）で観察される。内突起に近い弾性弦音ストランドとテンションニューロンに注意してください。直径の異なるいくつかの細胞体D特定の場所では、この図にも表示されます。テンション神経および運動神経ストレッチャーの全過程を図15に示す。 PDの神経の個々のニューロンを、4 -ジ-2-ASP（ 図16）とのCoCl 2（ 図17）の技術を用いて高コントラストバックフィルで示されている。高倍率での感覚終末は、支持scolopales（ 図16B）21,22,26の内側に見ることができる。 <img alt="図10" fo:content-width="5in" src="/files/ftp_upload/51050/51050fig10.jpg" width="500px" /> 図10は移動し、0℃で保持します 。ダイナミックニューロンは、運動中に焼成時の堅牢で、関節が開いたままにしている間、静的な感受性ニューロンからのスパイクが存在している。 <a href="https://www.jove.com/files/ftp_upload/51050/51050fig10highres.jpg" target="_blank">拡大画像を表示するには、ここをクリックしてください&lt;</a>/&gt;。 <img alt="図11" fo:content-width="5in" src="/files/ftp_upload/51050/51050fig11.jpg" width="500px" /> 図11。迅速、オープンで完全に伸び（90-0°）の位置に曲げられ終了開始 。 <a href="https://www.jove.com/files/ftp_upload/51050/51050fig11highres.jpg" target="_blank">拡大画像を表示するには、ここをクリックしてください</a> 。 <img alt="図12" fo:content-width="5in" src="/files/ftp_upload/51050/51050fig12.jpg" width="500px" /> 図12。のPDの神経から記録された細胞外スパイク 。関節が完全に伸びてからすぐに½に移動した位置を屈曲してから、まだ開催された。移動中に活動に気づくとき、静的な活動を低下させた。 <a href="https://www.jove.com/files/ftp_upload/51050/51050fig12highres.jpg" target="_blank">大きなIMAを見るにはここをクリックしてください</a>GE。 <img alt="図13" fo:content-width="5in" src="/files/ftp_upload/51050/51050fig13.jpg" width="500px" /> 図13。ジョイントを使用して開発された相対的な力は、完全に屈曲し、様々な周波数で刺激した（A）力変換器からの電圧-時間トレースは、各刺激周波数で示されている。 （B）パネルA内のトレースは、比較を容易にするために、異なる色で重ね合わせられる。運動神経は80 Hzで刺激したときの電気的活動（C）電圧-時間トレースは、張力神経から記録した。神経活動と比較して、刺激アーティファクトの規則的なパターンに注意してください。また、神経応答の様々な振幅の点に注意してください。 <a href="https://www.jove.com/files/ftp_upload/51050/51050fig13highres.jpg" target="_blank">拡大画像を表示するには、ここをクリックしてください</a> 。 図14。歩行脚の準備のメチレンブルー染色 。個々の細胞体は、弾性ストランドの中に突出彼らの感覚終末で表示されます。テンションニューロンが表示されている内突起があります。 <a href="https://www.jove.com/files/ftp_upload/51050/51050fig14highres.jpg" target="_blank">拡大画像を表示するには、ここをクリックしてください</a> 。 <img alt="図15" fo:content-width="5in" src="/files/ftp_upload/51050/51050fig15.jpg" width="500px" /> 図15。テンション神経末端部（赤い矢印）から生じると運動神経（緑の矢印）を結ぶ。 <a href="https://www.jove.com/files/ftp_upload/51050/51050fig15highres.jpg" target="_blank">拡大画像を表示するには、ここをクリックしてください</a> 。 4 -ジ-2-ASPによる癌ネギま内のPD神経の図16（A）バックフィル（B）の感覚終末の高倍率。 <a href="https://www.jove.com/files/ftp_upload/51050/51050fig16highres.jpg" target="_blank">拡大画像を表示するには、ここをクリックしてください</a> 。 <img alt="図17" fo:content-width="5in" src="/files/ftp_upload/51050/51050fig17.jpg" width="500px" /> 図17。のCoCl 2を充填し、処理したニューロン（A）。示す染色の準備のトレースされた輪郭線（B） <a href="https://www.jove.com/files/ftp_upload/51050/51050fig17highres.jpg" target="_blank">拡大画像を表示するには、ここをクリックしてください</a> 。 <table border="0" cellpadding="0" cellspacING = &quot;0&quot;幅= &quot;392&quot;&gt; <colgroup><col /><col /></colgroup><tbody><tr height="21"><td height="21" style="height:21px;width:235px;"> 生理食塩水 </td><td style="width:157px;"> g / Lの </td></tr><tr height="20"><td height="20" style="height:20px;width:235px;"> NaClを</td><td style="width:157px;"> 27.29 </td></tr><tr height="20"><td height="20" style="height:20px;width:235px;">塩化カリウム</td><td style="width:157px;"> 0.81 </td></tr><tr height="26"><td height="26" style="height:26px;width:235px;">硫酸マグネシウム4•7H 2 O </td><td style="width:157px;"> 4.81 </td></tr><tr height="26"><td height="26" style="height:26px;width:235px;">のCaCl 2•2H 2 O </td><td style="width:157px;"> 1.85 </td></tr><tr height="26"><td height="26" style="height:26px;width:235px;">のNa 2 SO 4•10H 2 O </td><td style="width:157px;"> 0.97 </td></tr><tr height=&quot;20&quot;&gt; <td height="20" style="height:20px;width:235px;">デキストロース（D-グルコース） </td><td style="width:157px;"> 1.982 </td></tr><tr height="20"><td height="20" style="height:20px;width:235px;"> HEPES酸</td><td style="width:157px;"> 0.476 </td></tr><tr height="20"><td height="20" style="height:20px;width:235px;"> HEPES塩</td><td style="width:157px;"> 2.08 </td></tr><tr height="40"><td height="40" style="height:40px;width:235px;"> NaOHまたはHClでpHを8.1に調整し</td><td style="width:157px;"></td></tr></tbody></table> 表1。カニの生理食塩水のレシピ。

Watch this Scientific Journal Video about Proprioception and Tension Receptors in Crab Limbs: Student Laboratory Exercises at JoVE.com

カニの手足の固有受容体と張力受容体 |神経科学 |ビデオ

Proprioception and Tension Receptors in Crab Limbs: Student Laboratory Exercises

生理学的および解剖学的技法は甲殻類の歩行手足の関節の固有受容および筋肉の緊張受容体の機能および構造に対処することが実証されている。

カニ手足の固有感覚と緊張受容体：学生実験室の練習

The primary purpose of these procedures is to demonstrate for teaching and research purposes how to record the activity of living primary sensory neurons ...

proprioception-tension-receptors-crab-limbs-student-laboratory

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25.1K ビュー.  University of Kentucky. 生理学的および解剖学的技法は甲殻類の歩行手足の関節の固有受容および筋肉の緊張受容体の機能および構造に対処することが実証されている。 

ビデオ: Proprioception and Tension Receptors in Crab Limbs: Student Laboratory Exercises

Spectral Confocal Imaging of Fluorescently tagged Nicotinic Receptors in Knock-in Mice with Chronic Nicotine Administration

Ligand-gated ion channels in the central nervous system (CNS) are implicated in numerous conditions with serious medical and social consequences. For instance, addiction to nicotine via tobacco smoking is a leading cause of premature death worldwide (World Health Organization) and is likely caused by an alteration of ion channel distribution in the brain1. Chronic nicotine exposure in both rodents and humans results in increased numbers of nicotinic acetylcholine receptors (nAChRs) in brain tissue1-3. Similarly, alterations in the glutamatergic GluN1 or GluA1 channels have been implicated in triggering sensitization to other addictive drugs such as cocaine, amphetamines and opiates4-6.

Consequently, the ability to map and quantify distribution and expression patterns of specific ion channels is critically important to understanding the mechanisms of addiction. The study of brain region-specific effects of individual drugs was advanced by the advent of techniques such as radioactive ligands. However, the low spatial resolution of radioactive ligand binding prevents the ability to quantify ligand-gated ion channels in specific subtypes of neurons. 

Genetically encoded fluorescent reporters, such as green fluorescent protein (GFP) and its many color variants, have revolutionized the field of biology7.By genetically tagging a fluorescent reporter to an endogenous protein one can visualize proteins in vivo7-10. One advantage of fluorescently tagging proteins with a probe is the elimination of antibody use, which have issues of nonspecificity and accessibility to the target protein. We have used this strategy to fluorescently label nAChRs, which enabled the study of receptor assembly using F&ouml;rster Resonance Energy Transfer (FRET) in transfected cultured cells11.More recently, we have used the knock-in approach to engineer mice with yellow fluorescent protein tagged &alpha;4 nAChR subunits (&alpha;4YFP), enabling precise quantification of the receptor ex vivo at submicrometer resolution in CNS neurons via spectral confocal microscopy12. The targeted fluorescent knock-in mutation is incorporated in the endogenous locus and under control of its native promoter, producing normal levels of expression and regulation of the receptor when compared to untagged receptors in wildtype mice. This knock-in approach can be extended to fluorescently tag other ion channels and offers a powerful approach of visualizing and quantifying receptors in the CNS.

In this paper we describe a methodology to quantify changes in nAChR expression in specific CNS neurons after exposure to chronic nicotine. Our methods include mini-osmotic pump implantation, intracardiac perfusion fixation, imaging and analysis of fluorescently tagged nicotinic receptor subunits from &alpha;4YFP knock-in mice (Fig. 1). We have optimized the fixation technique to minimize autofluorescence from fixed brain tissue.We describe in detail our imaging methodology using a spectral confocal microscope in conjunction with a linear spectral unmixing algorithm to subtract autofluoresent signal in order to accurately obtain &alpha;4YFP fluorescence signal. Finally, we show results of chronic nicotine-induced upregulation of &alpha;4YFP receptors in the medial perforant path of the hippocampus.

We have developed a novel technique of quantifying nicotinic acetylcholine receptor changes within subcellular regions of specific subtypes of CNS neurons to better understand the mechanisms of nicotine addiction by using a combination of approaches including fluorescent protein tagging of the receptor using the knock-in approach and spectral confocal imaging.

慢性ニコチン投与によるマウスノックインで蛍光標識ニコチン性受容体のスペクトル共焦点イメージング

Ligand-gated ion channels in the central nervous system (CNS) are implicated in numerous conditions with serious medical and social consequences.  For ...

神経科学

Use of pHluorin to Assess the Dynamics of Axon Guidance Receptors in Cell Culture and in the Chick Embryo

During development, axon guidance receptors play a crucial role in regulating axons sensitivity to both attractive and repulsive cues. Indeed, activation of the guidance receptors is the first step of the signaling mechanisms allowing axon tips, the growth cones, to respond to the ligands. As such, the modulation of their availability at the cell surface is one of the mechanisms that participate in setting the growth cone sensitivity. We describe here a method to precisely visualize the spatio-temporal cell surface dynamics of an axon guidance receptor both in vitro and in vivo in the developing chick spinal cord. We took advantage of the pH-dependent fluorescence property of a green fluorescent protein (GFP) variant to specifically detect the fraction of the axon guidance receptor that is addressed to the plasma membrane. We first describe the in vitro validation of such pH-dependent constructs and we further detail their use in vivo, in the chick spinal chord, to assess the spatio-temporal dynamics of the axon guidance receptor of interest.

We describe here the use of a pH-sensitive green fluorescent protein variant, pHluorin, to study the spatio-temporal dynamics of axon guidance receptors trafficking at the cell surface.&#160;The pHluorin-tagged receptor is expressed both in cell culture and in vivo, using electroporation of the chick embryo.

細胞培養およびひよこ胚における軸索誘導受容体のダイナミクス評価におけるpHluorinの使用

During development, axon guidance receptors play a crucial role in regulating axons sensitivity to both attractive and repulsive cues. Indeed, activation ...

Inducing Plasticity of Astrocytic Receptors by Manipulation of Neuronal Firing Rates

Close to two decades of research has established that astrocytes in situ and in vivo express numerous G protein-coupled receptors (GPCRs) that can be stimulated by neuronally-released transmitter. However, the ability of astrocytic receptors to exhibit plasticity in response to changes in neuronal activity has received little attention. Here we describe a model system that can be used to globally scale up or down astrocytic group I metabotropic glutamate receptors (mGluRs) in acute brain slices. Included are methods on how to prepare parasagittal hippocampal slices, construct chambers suitable for long-term slice incubation, bidirectionally manipulate neuronal action potential frequency, load astrocytes and astrocyte processes with fluorescent Ca2+ indicator, and measure changes in astrocytic Gq GPCR activity by recording spontaneous and evoked astrocyte Ca2+ events using confocal microscopy. In essence, a &#8220;calcium roadmap&#8221; is provided for how to measure plasticity of astrocytic Gq GPCRs. Applications of the technique for study of astrocytes are discussed. Having an understanding of how astrocytic receptor signaling is affected by changes in neuronal activity has important implications for both normal synaptic function as well as processes underlying neurological disorders and neurodegenerative disease.

Here we describe an adaptation of protocols used to induce homeostatic plasticity in neurons for the study of plasticity of astrocytic G protein-coupled receptors. Recently used to examine changes in astrocytic group I mGluRs in juvenile mice, the method can be applied to measure scaling of various astrocytic GPCRs, in tissue from adult mice in situ and in vivo, and to gain a better appreciation of the sensitivity of astrocytic receptors to changes in neuronal activity.

神経発火率を操作することによってアストロサイト受容体の可塑性を誘導すること

Close to two decades of research has established that astrocytes in situ and in vivo express numerous G protein-coupled receptors (GPCRs) that can be ...

Inhibitory Synapse Formation in a Co-culture Model Incorporating GABAergic Medium Spiny Neurons and HEK293 Cells Stably Expressing GABAA Receptors

Inhibitory neurons act in the central nervous system to regulate the dynamics and spatio-temporal co-ordination of neuronal networks. GABA (&#947;-aminobutyric acid) is the predominant inhibitory neurotransmitter in the brain. It is released from the presynaptic terminals of inhibitory neurons within highly specialized intercellular junctions known as synapses, where it binds to GABAA receptors (GABAARs) present at the plasma membrane of the synapse-receiving, postsynaptic neurons. Activation of these GABA-gated ion channels leads to influx of chloride resulting in postsynaptic potential changes that decrease the probability that these neurons will generate action potentials. 
During development, diverse types of inhibitory neurons with distinct morphological, electrophysiological and neurochemical characteristics have the ability to recognize their target neurons and form synapses which incorporate specific GABAARs subtypes. This principle of selective innervation of neuronal targets raises the question as to how the appropriate synaptic partners identify each other. 
To elucidate the underlying molecular mechanisms, a novel in vitro co-culture model system was established, in which medium spiny GABAergic neurons, a highly homogenous population of neurons isolated from the embryonic striatum, were cultured with stably transfected HEK293 cell lines that express different GABAAR subtypes. Synapses form rapidly, efficiently and selectively in this system, and are easily accessible for quantification. Our results indicate that various GABAAR subtypes differ in their ability to promote synapse formation, suggesting that this reduced in vitro model system can be used to reproduce, at least in part, the in vivo conditions required for the recognition of the appropriate synaptic partners and formation of specific synapses. Here the protocols for culturing the medium spiny neurons and generating HEK293 cells lines expressing GABAARs are first described, followed by detailed instructions on how to combine these two cell types in co-culture and analyze the formation of synaptic contacts.

Inhibitory Synapse Formation in a Co-culture Model Incorporating GABAergic Medium Spiny Neurons and HEK293 Cells Stably Expressing GABAA Receptors

The molecular mechanisms that co-ordinate the formation of inhibitory GABAergic synapses during ontogeny are largely unknown. To study these processes,we have developed a co-culture model system which incorporates embryonic medium spiny GABAergic neurons cultured together with stably transfected human embryonic kidney 293 (HEK293) cells expressing functional GABAA receptors.

共培養モデルの組み込みGABA作動性中型有棘ニューロンおよびHEK293細胞における抑制性シナプスの形成を安定的にGABAを表現<sub&gt; A</sub&gt;受容体

Inhibitory neurons act in the central nervous system to regulate the dynamics and spatio-temporal co-ordination of neuronal networks. GABA ...

High-Resolution Quantitative Immunogold Analysis of Membrane Receptors at Retinal Ribbon Synapses

Retinal ganglion cells (RGCs) receive excitatory glutamatergic input from bipolar cells. Synaptic excitation of RGCs is mediated postsynaptically by NMDA receptors (NMDARs) and AMPA receptors (AMPARs). Physiological data have indicated that glutamate receptors at RGCs are expressed not only in postsynaptic but also in perisynaptic or extrasynaptic membrane compartments. However, precise anatomical locations for glutamate receptors at RGC synapses have not been determined. Although a high-resolution quantitative analysis of glutamate receptors at central synapses is widely employed, this approach has had only limited success in the retina. We developed a postembedding immunogold method for analysis of membrane receptors, making it possible to estimate the number, density and variability of these receptors at retinal ribbon synapses. Here we describe the tools, reagents, and the practical steps that are needed for: 1) successful preparation of retinal fixation, 2) freeze-substitution, 3) postembedding immunogold electron microscope (EM) immunocytochemistry and, 4) quantitative visualization of glutamate receptors at ribbon synapses.

The postembedding immunogold method is one of the most effective ways to provide high-resolution analyses of the subcellular localization of specific molecules. Here we describe a protocol to quantitatively analyze glutamate receptors at retinal ribbon synapses.

網膜リボンシナプスにおける膜受容体の高分解能定量的イムノゴールド分析

Retinal ganglion cells (RGCs) receive excitatory glutamatergic input from bipolar cells. Synaptic excitation of RGCs is mediated postsynaptically by NMDA ...

Quantification of Filamentous Actin (F-actin) Puncta in Rat Cortical Neurons

Filamentous actin protein (F-actin) plays a major role in spinogenesis, synaptic plasticity, and synaptic stability. Changes in dendritic F-actin rich structures suggest alterations in synaptic integrity and connectivity. Here we provide a detailed protocol for culturing primary rat cortical neurons, Phalloidin staining for F-actin puncta, and subsequent quantification techniques. First, the frontal cortex of E18 rat embryos are dissociated into low-density cell culture, then the neurons grown in vitro for at least 12-14 days. Following experimental treatment, the cortical neurons are stained with AlexaFluor 488 Phalloidin (to label the dendritic F-actin puncta) and microtubule-associated protein 2 (MAP2; to validate the neuronal cells and dendritic integrity). Finally, specialized software is used to analyze and quantify randomly selected neuronal dendrites. F-actin rich structures are identified on second order dendritic branches (length range 25-75 &#181;m) with continuous MAP2 immunofluorescence. The protocol presented here will be a useful method for investigating changes in dendritic synapse structures subsequent to experimental treatments.

Quantification of Filamentous Actin F-actin Puncta in Rat Cortical Neurons

Filamentous actin (F-actin) plays an important role in spinogenesis, synaptic plasticity, and synaptic stability. Quantification of F-actin puncta is therefore a useful tool to study the integrity of synaptic structures. This protocol describes the procedures of quantifying F-actin puncta labeled with Phalloidin in low-density primary cortical neuronal cultures.

ラット皮質ニューロンにおける繊維状アクチン（F-アクチン）涙点の定量化

Filamentous actin protein (F-actin) plays a major role in spinogenesis, synaptic plasticity, and synaptic stability. Changes in dendritic F-actin rich ...

Combined Optogenetic and Freeze-fracture Replica Immunolabeling to Examine Input-specific Arrangement of Glutamate Receptors in the Mouse Amygdala

Freeze-fracture electron microscopy has been a major technique in ultrastructural research for over 40 years. However, the lack of effective means to study the molecular composition of membranes produced a significant decline in its use. Recently, there has been a major revival in freeze-fracture electron microscopy thanks to the development of effective ways to reveal integral membrane proteins by immunogold labeling. One of these methods is known as detergent-solubilized Freeze-fracture Replica Immunolabeling (FRIL).
The combination of the FRIL technique with optogenetics allows a correlated analysis of the structural and functional properties of central synapses. Using this approach it is possible to identify and characterize both pre- and postsynaptic neurons by their respective expression of a tagged channelrhodopsin and specific molecular markers. The distinctive appearance of the postsynaptic membrane specialization of glutamatergic synapses further allows, upon labeling of ionotropic glutamate receptors, to quantify and analyze the intrasynaptic distribution of these receptors. Here, we give a step-by-step description of the procedures required to prepare paired replicas and how to immunolabel them. We will also discuss the caveats and limitations of the FRIL technique, in particular those associated with potential sampling biases. The high reproducibility and versatility of the FRIL technique, when combined with optogenetics, offers a very powerful approach for the characterization of different aspects of synaptic transmission at identified neuronal microcircuits in the brain.
Here, we provide an example how this approach was used to gain insights into structure-function relationships of excitatory synapses at neurons of the intercalated cell masses of the mouse amygdala. In particular, we have investigated the expression of ionotropic glutamate receptors at identified inputs originated from the thalamic posterior intralaminar and medial geniculate nuclei. These synapses were shown to relay sensory information relevant for fear learning and to undergo plastic changes upon fear conditioning.

This article illustrates how the expression of neurotransmitter receptors can be quantified and the pattern analyzed at synapses with identified pre and postsynaptic elements using a combination of viral transduction of optogenetic tools and the freeze-fracture replica immunolabeling technique.

複合光遺伝学および凍結破砕レプリカ免疫標識をマウス扁桃体におけるグルタミン酸受容体の入力固有の配置を検討すること

Freeze-fracture electron microscopy has been a major technique in ultrastructural research for over 40 years. However, the lack of effective means to ...

Receptor Autoradiography Protocol for the Localized Visualization of Angiotensin II Receptors

This protocol describes receptor binding patterns for Angiotensin II (Ang II) in the rat brain using a radioligand specific for Ang II receptors to perform receptor autoradiographic mapping.
Tissue specimens are harvested and stored at -80 &#176;C. A cryostat is used to coronally section the tissue (brain) and thaw-mount the sections onto charged slides. The slide-mounted tissue sections are incubated in 125I-SI-Ang II to radiolabel Ang II receptors. Adjacent slides are separated into two sets: &#39;non-specific binding&#39; (NSP) in the presence of a receptor saturating concentration of non-radiolabeled Ang II, or an AT1 Ang II receptor subtype (AT1R) selective Ang II receptor antagonist, and &#39;total binding&#39; with no AT1R antagonist. A saturating concentration of AT2 Ang II receptor subtype (AT2R) antagonist (PD123319, 10 &#181;M) is also present in the incubation buffer to limit 125I-SI-Ang II binding to the AT1R subtype. During a 30 min pre-incubation at ~22 &#176;C, NSP slides are exposed to 10 &#181;M PD123319 and losartan, while &#39;total binding&#39; slides are exposed to 10 &#181;M PD123319. Slides are then incubated with 125I-SI-Ang II in the presence of PD123319 for &#39;total binding&#39;, and PD123319 and losartan for NSP in assay buffer, followed by several &#39;washes&#39; in buffer, and water to remove salt and non-specifically bound radioligand. The slides are dried using blow-dryers, then exposed to autoradiography film using a specialized film and cassette. The film is developed and the images are scanned into a computer for visual and quantitative densitometry using a proprietary imaging system and a spreadsheet. An additional set of slides are thionin-stained for histological comparisons.
The advantage of using receptor autoradiography is the ability to visualize Ang II receptors in situ, within a section of a tissue specimen, and anatomically identify the region of the tissue by comparing it to an adjacent histological reference section.

Here we present a protocol to describe the localization of angiotensin II Type 1 receptors in the rat brain by quantitative, densitometric, in vitro receptor autoradiography using an iodine-125 labeled analog of angiotensin II.

アンジオテンシンII受容体のローカライズされた可視化のための受容体オートラジオグラフィープロトコル

This protocol describes receptor binding patterns for Angiotensin II (Ang II) in the rat brain using a radioligand specific for Ang II receptors to ...

Assessment of Dopaminergic Homeostasis in Mice by Use of High-performance Liquid Chromatography Analysis and Synaptosomal Dopamine Uptake

Dopamine (DA) is a modulatory neurotransmitter controlling motor activity, reward processes and cognitive function. Impairment of dopaminergic (DAergic) neurotransmission is strongly associated with several central nervous system-associated diseases such as Parkinson&#39;s disease, attention-deficit-hyperactivity disorder and drug addiction1,2,3,4. Delineating disease mechanisms involving DA imbalance is critically dependent on animal models to mimic aspects of the diseases, and thus protocols that assess specific parts of the DA homeostasis are important to provide novel insights and possible therapeutic targets for these diseases.
Here, we present two useful experimental protocols that when combined provide a functional read-out of the DAergic system in mice. Biochemical and functional parameters on DA homeostasis are obtained through assessment of DA levels and dopamine transporter (DAT) functionality5. When investigating the DA system, the ability to reliably measure endogenous levels of DA from adult brain is essential. Therefore, we present how to perform high-performance liquid chromatography (HPLC) on brain tissue from mice to determine levels of DA. We perform the experiment on tissue from dorsal striatum (dStr) and nucleus accumbens (NAc), but the method is also suitable for other DA-innervated brain areas.
DAT is essential for reuptake of DA into the presynaptic terminal, thereby controlling the temporal and spatial activity of released DA. Knowing the levels and functionality of DAT in the striatum is of major importance when assessing DA homeostasis. Here, we provide a protocol that allows to simultaneously deduce information on surface levels and function using a synaptosomal6 DA uptake assay.
Current methods combined with standard immunoblotting protocols provide the researcher with relevant tools to characterize the DAergic system.

Synaptosomal dopamine uptake and high-performance liquid chromatography analysis represent experimental tools to investigate dopamine homeostasis in mice by assessing the function of the dopamine transporter and levels of dopamine in striatal tissue, respectively. Here we present protocols to measure dopamine tissue content and assess the functionality of the dopamine transporter.

高速液体クロマトグラフィー分析とシナプトソーム ドーパミンによるマウスにおけるドーパミン作動性恒常性の評価

Dopamine (DA) is a modulatory neurotransmitter controlling motor activity, reward processes and cognitive function. Impairment of dopaminergic (DAergic) ...

Adaptation of Microelectrode Array Technology for the Study of Anesthesia-induced Neurotoxicity in the Intact Piglet Brain

Every year, millions of children undergo anesthesia for a multitude of procedures. However, studies in both animals and humans have called into question the safety of anesthesia in children, implicating anesthetics as potentially toxic to the brain in development. To date, no studies have successfully elucidated the mechanism(s) by which anesthesia may be neurotoxic. Animal studies allow investigation of such mechanisms, and neonatal piglets represent an excellent model to study these effects due to their striking developmental similarities to the human brain.
This protocol adapts the use of enzyme-based microelectrode array (MEA) technology as a novel way to study the mechanism(s) of anesthesia-induced neurotoxicity (AIN). MEAs enable real-time monitoring of in vivo neurotransmitter activity and offer exceptional temporal and spatial resolution. It is hypothesized that anesthetic neurotoxicity is caused in part by glutamate dysregulation and MEAs offer a method to measure glutamate. The novel implementation of MEA technology in a piglet model presents a unique opportunity for the study of AIN.

This study explores the novel use of enzyme-based microelectrode array (MEA) technology to monitor in vivo neurotransmitter activity in piglets. The hypothesis was that glutamate dysregulation contributes to the mechanism of anesthetic neurotoxicity. Here, we present a protocol to adapt MEA technology to study the mechanism of anesthesia-induced neurotoxicity.

そのまま子豚脳内麻酔神経毒性の研究のための微小電極アレイ技術の適応

Every year, millions of children undergo anesthesia for a multitude of procedures. However, studies in both animals and humans have called into question ...