Name: probing the roles of physical forces in early chick embryonic morphogenesis
Uploaded: 2018-06-05T13:00:00
Description: Watch this Scientific Journal Video about Probing the Roles of Physical Forces in Early Chick Embryonic Morphogenesis at JoVE.com

Z.C. は、ダートマス スタートアップ資金とブランコ ヴァイスシュヴァルツ-社会科学フェローシップ、ETH チューリッヒによって管理されてからのサポートを認めています。著者は、本ラリー A. テーバー、Benjamen A. Filas、Qiaohang 郭と役に立つ議論のため Yunfei 市として匿名の校閲者のコメントをありがとうございます。この資料は、国立科学財団大学院研究奨学金助成号下に支えられて仕事に基づいていますDGE-1313911。意見、所見、結論または本資料に示した推奨事項著者 (s) のものし、国立科学財団の見解を必ずしも反映されません。

1. 培養媒体の作製
<ol><li>4.5 G/L グルコース、炭酸水素ナトリウム、L-グルタミンと文化メディアの拠点として 0.5 L ボトル ダルベッコ変更されたワシの媒体 (DMEM) を使用します。</li>
	<li>滅菌の層流フードの DMEM の 0.5 L に抗生物質の 10 mL を追加します。</li>
	<li>滅菌ピペットを使用して、滅菌 50 mL の円錐管に DMEM 抗生物質溶液 50 mL を転送します。</li>
	<li>ニワトリ血清の 50 mL を無菌フード 0.5...

<ol>
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M., Alvarez-Bolado, G., Treichel, D., Gruss, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rotatin+is+a+novel+gene+required+for+axial+rotation+and+left-right+specification+in+mouse+embryos.">Rotatin is a novel gene required for axial rotation and left-right specification in mouse embryos.</a> Mech. Dev. 113, 15-28 (2002).</li><li>Yalcin, H. C., Shekhar, A., Rane, A. a., Butcher, J. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+ex-ovo+chicken+embryo+culture+system+suitable+for+imaging+and+microsurgery+applications.">An ex-ovo chicken embryo culture system suitable for imaging and microsurgery applications.</a> J. Vis. Exp. (44), (2010).</li><li>Chapman, S. C., Collignon, J., Schoenwolf, G. 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物理現象形態26,27,28,29,30機械の調整と共に、特定の機械的メカニズムに不可欠な役割を果たす一方、分子機構は、主として未踏のままです。腹側の曲げと原始的な脳の右ねじり初期胚形態形成18,31,32<su...

現代の発生生物学研究は主として分子遺伝学1,2,3,4,5,6の観点から理解開発に焦点を当ててください。,7,8,9,10,11,12,13それは知られている物理現象が形態形成の中心的な役割を再生または生物の生成フォーム14,15,16,17;。しかし、開発の機械的なメカニズムは特定のまま主巧まない。原始的な脳管後ハンバーガー ハミルトン ステージ 11 (HH 11)18胚の形に貢献する 2 つの主要なプロセスの右の捩り及び腹側曲げ19,20を変更します。特に、胎生期の脳のねじれの開発の基になる物理的なメカニズムは不完全な理解のままになります。
ニワトリ胚の胚のねじりは開発で左右 (L R) 非対称性の最も早い形成イベントです。左右非対称のプロセスを摂動21内臓逆位、異性、または内臓などの先天異常が発生します。 初期の胎生期脳開発中に機械力を特徴付ける物理モデリングとex ovo実験22,23を組み合わせたプロトコルをご紹介します。提示方法の目標は、脳のねじりおよび初期開発12の中にねじりの程度に影響する要因を担当の機械力を識別するためにです。卵黄膜 (VM) が胚の背側を拘束実験観察に基づいて、VM が発展途上の脳のねじりを誘導するために必要な力を提供することを行った。したがって、この方法では、脳のねじりに及ぼす影響を調べる脳領域をカバーする VM の一部を削除しました。さらに、流体の表面張力を適用する方法は、VM の力学的役割を確認し、脳の捻転は、以前行われていないために必要な力の見積もりを提供する使用されました。萌芽期の形態形成時に力を測定は、やりがいのある仕事です。特に、先駆的な研究では、Campàs と同僚の24は、注入された微小油滴中を使用して携帯電話の応力を定量化する手法を開発しました。それにもかかわらず、このメソッドは、軍組織または個体レベルでのプローブには適用されませんので、細胞レベルで力を測定するに限られていた。本稿で提示されたプロトコルは、部分的にこのギャップを埋めるために開発されました。

<table border="0" cellpadding="0" cellspacing="0" width="751">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:288px;">Fertilized Specific pathogen-free White Leghorn chicken eggs</td>
			<td style="width:177px;">Charles River</td>
			<td style="width:119px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:288px;">Optical Coherent Tomography Microscope</td>
			<td style="width:177px;">Thorlabs</td>
			<td style="width:119px;">GAN220C1</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:288px;">Silicone elastomer</td>
			<td style="width:177px;">Smooth-On, Inc.</td>
			<td style="width:119px;">EcoFlex 00-50</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:288px;">Dissecting microscope</td>
			<td style="width:177px;">Leica</td>
			<td style="width:119px;">MZ8</td>
			<td></td>
		</tr>
		<tr height="39">
			<td height="39" style="height:39px;width:288px;">Dulbecco&#8217;s Modified Eagle&#8217;s Medium (DMEM)</td>
			<td style="width:177px;">Lonza</td>
			<td style="width:119px;">12-604F</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:288px;">Antibiotics</td>
			<td style="width:177px;">Sigma</td>
			<td style="width:119px;">P4083</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:288px;">Chick serum</td>
			<td style="width:177px;">Sigma</td>
			<td style="width:119px;">C5405</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:288px;">Micropipette puller</td>
			<td style="width:177px;">Sutter Instrument</td>
			<td style="width:119px;">Model P-30</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:288px;">Filter paper</td>
			<td style="width:177px;">Whatman</td>
			<td style="width:119px;">5202-110</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:288px;">Phosphate buffered saline (PBS)</td>
			<td style="width:177px;">Corning</td>
			<td style="width:119px;">21-040-CV</td>
			<td></td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:288px;">Comsol MultiPhysics</td>
			<td style="width:177px;">Comsol</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:288px;">3D computer graphics software</td>
			<td style="width:177px;">Rhino 5</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:288px;">Microscope attached with OCT</td>
			<td style="width:177px;">Nikon&#160;</td>
			<td style="width:119px;">FN1</td>
			<td></td>
		</tr>
		<tr height="18">
			<td height="18" style="height:18px;width:288px;">Digital single-lens reflex camera</td>
			<td style="width:177px;">EOS&#160;</td>
			<td style="width:119px;">Rebel T3i</td>
			<td></td>
		</tr>
	</tbody>
</table>

probing the roles of physical forces in early chick embryonic morphogenesis

萌芽期の開発、分子遺伝学の観点から検討する伝統的が形態形成における力学の基本的な重要性がますます認識になります。特に、鶏胚心と脳管、抜本的な形態学的変化を受けることを開発する際、形態形成における物理的な力の役割を研究する首相候補の中では。進歩的な腹曲げと管状初期胚の脳の右ねじりは、ニワトリ胚の器官レベルの左右非対称性の初期の段階で起こる。卵黄膜 (VM) は、胚の背側を固定され、脳の発達のねじりを誘導するために必要な力を提供することに関与しています。新しいex ovo実験と物理脳ねじりの力学を識別するためにモデリングの組み合わせをご紹介します。ハンバーガー ハミルトン 11 段階で胚が収穫され、培養ovo ex (メディアで)。VM では、引っ張られたキャピラリー チューブを使用して削除されます後。流体のレベルを制御して胚を液体空気インターフェイスに服従させることで、メディアの流体の表面張力は、VM の力学的役割を交換する使用できます。マイクロサージャリー実験は脳ねじりのカイラリティの結果の変化を見つける心の位置を変更するも行った。このプロトコルからの結果は、運転形態形成の力学の基本的な役割を示しています。

本研究では HH11 で胚の VM は胸部の純曲げを前方から削除されました。OCT システムによる胚をイメージしました。この段階で脳管のねじりはありません (図 1 a) を開始しました。HH15 16 に産み落とされていた後削除の VM を持つ胚は低脳管のねじり、約 35 度 (図 1 b) 90 度前後のねじりを表わす胚をコントロールと比較して展示...

Watch this Scientific Journal Video about Probing the Roles of Physical Forces in Early Chick Embryonic Morphogenesis at JoVE.com

Probing the Roles of Physical Forces in Early Chick Embryonic Morphogenesis

初期ニワトリ胚形態形成における物理的な力の役割を探る

ここでは、一連の新しいex ovo実験と初期ニワトリ胚の脳ねじり中に形態形成の仕組みを勉強するための物理的なモデリング アプローチを紹介プロトコルを提案します。

Embryonic development is traditionally studied from the perspective of biomolecular genetics, but the fundamental importance of mechanics in morphogenesis ...

Research

JoVE Journal

Bioengineering

Multiparametric Optical Mapping of the Langendorff-perfused Rabbit Heart

ランゲンドルフ灌流ウサギの心臓のMultiparametric光学マッピング

Optical imaging and fluorescent probes have significantly advanced research methodology in the field of cardiac electrophysiology in ways that could not ...

Remote Magnetic Actuation of Micrometric Probes for in situ 3D Mapping of Bacterial Biofilm Physical Properties

Remote Magnetic Actuation of Micrometric Probes for in situ 3D Mapping of Bacterial Biofilm Physical Properties

のためのマイクロメトリックプローブの遠隔磁気作動<em&gt;その場での</em細菌バイオフィルム物性&gt; 3Dマッピング

Bacterial adhesion and growth on interfaces lead to the formation of three-dimensional heterogeneous structures so-called biofilms. The cells dwelling in ...

Quantification of Global Diastolic Function by Kinematic Modeling-based Analysis of Transmitral Flow via the Parametrized Diastolic Filling Formalism

パラメータ化拡張期充満フォーマリズムを経由Transmitral流れの運動学的モデルベースの​​解析によるグローバル拡張期機能の定量化

Quantitative cardiac function assessment remains a challenge for physiologists and clinicians.&#160;Although historically invasive methods have comprised ...

A Protocol for Using F&#246;rster Resonance Energy Transfer (FRET)-force Biosensors to Measure Mechanical Forces across the Nuclear LINC Complex

A Protocol for Using FÃƒÆ’Ã†'Ãƒâ€šÃ‚Â¶rster Resonance Energy Transfer FRET-force Biosensors to Measure Mechanical Forces across the Nuclear LINC Complex

原子力LINCコンプレックス全体に機械的な力を測定するために、蛍光共鳴エネルギー移動（FRET）-forceのバイオセンサーを使用するためのプロトコル

The LINC complex has been hypothesized to be the critical structure that mediates the transfer of mechanical forces from the cytoskeleton to the nucleus. ...

A High-throughput Cell Microarray Platform for Correlative Analysis of Cell Differentiation and Traction Forces

細胞の分化やトラクション軍の相関的分析のためのハイスループット細胞マイクロアレイプラットフォーム

Microfabricated cellular microarrays, which consist of contact-printed combinations of biomolecules on an elastic hydrogel surface, provide a tightly ...

An In Vitro Organ Culture Model of the Murine Intervertebral Disc

An In Vitro Organ Culture Model of the Murine Intervertebral Disc

アン<em&gt;インビトロ</emマウス椎間板の&gt;器官培養モデル

Intervertebral disc (IVD) degeneration is a significant contributor to low back pain. The IVD is a fibrocartilaginous joint that serves to transmit and ...

Improved 3D Hydrogel Cultures of Primary Glial Cells for In Vitro Modelling of Neuroinflammation

Improved 3D Hydrogel Cultures of Primary Glial Cells for In Vitro Modelling of Neuroinflammation

プライマリのグリア細胞の in Vitro Neuroinflammation のモデリングのための三次元ハイドロゲル文化を改善

In the central nervous system, numerous acute injuries and neurodegenerative disorders, as well as implanted devices or biomaterials engineered to enhance ...

A Tripeptide-Stabilized Nanoemulsion of Oleic Acid

オレイン酸のトリペプチド安定ナノエマルション

We describe a method to produce a nanoemulsion composed of an oleic acids-Pt(II) core and a lysine-tyrosine-phenylalanine (KYF) coating (KYF-Pt-NE). The ...

Patterning the Geometry of Human Embryonic Stem Cell Colonies on Compliant Substrates to Control Tissue-Level Mechanics

組織レベルの力学を制御するための準拠基板上のヒト胚性幹細胞コロニーの幾何学的パターン化

Human embryonic stem cells demonstrate a unique ability to respond to morphogens in vitro by self-organizing patterns of cell fate specification that ...

Application of Atomic Force Microscopy to Detect Early Osteoarthritis

早期変形性関節症の検出に対する原子間力顕微鏡の応用

Biomechanical properties of cells and tissues not only regulate their shape and function but are also crucial for maintaining their vitality. Changes in ...