この作品は、健康補助金の国立研究所、R01 AG033620（BG）で、ロバート·ドレイパー技術革新資金（DAW）によってサポートされていました。

HITデバイスの1建設<ol><li> 2つのクランプと4本のネジ（ 図1A）を使用して、ボードにスプリングを取り付けます。ボードの幅を基準にクランプを中心とボードの縁と外側のクランプ面一に互いに対してそれらを突き合わせ。クランプを取り付ける前に、バネの上にしっかりとフィットするようにペンチを使用してそれらを曲げます。 注：HITデバイスを構築するために必要な材料の説明については、表1を参照してください。春のクランプ端部は、基板の端から1/8インチ（3.2ミリメートル）であるべきであり、自由端が3/4インチ（19ミリメートル）て、ボード上に拡張する必要があります。それは板の長さと平行に位置するようにバネを調整します。 </li><li>ベルクロループの粘着テープで一度周りのばねの自由端を包みます。マジックテープの外縁部は、スプリングの端部と同一平面である必要があります。それはCREによってバネに、バイアルを固定するために使用されるので、マジックテープが重要ですタイトな圧縮フィッティングをレーティング。ベルクロは、多くのバイアルを短時間で処理することができるように、バイアル瓶の容易な接続及び取り外しを可能にします。 </li><li>タイトな木の板に対して、中心に、逆さまアイスバケットカバーを置きます。オリエント長辺は、ボードの幅と平行になるようアイスバケットカバーの隆起領域。バイアルをバネに取り付けられたとき、スプリングがボード上に平らにしないようにアイスバケットの隆起領域は、木の板よりも1/2インチ（13ミリメートル）高いことに注意してください。 </li><li>アイスバケットカバーは基板と対象物の間に押し込まれ、動かないように、壁などの固定オブジェクトに対して、装置全体をスライドさせます。 </li><li>テープ段ボールの底に紙分度器は、バイアルトレイを飛行し、それが完全に垂直位置に引き戻されたときに90°のマークは、ばねと整列されるように、基板の長さに対して、エッジの上に立っています。 </li></ol> 2。HIT装置の動作<ol><li>空のバイアルにハエやタイトフィットのコットンボールを使用して、バイアルを栓-anesthetized 1と60 CO 2との間に配置します。 </li><li>それが下から1インチ（2.5センチ）になるまでバイアルにコットンボールを押してバイアルの底1インチ（2.5センチ）にハエを閉じ込めます。これは、1インチ（2.5センチ）のマークでバイアル上に線を描画するために有用です。バイアルのより大きなまたはより小さな領域にハエを閉じ込めることは表現型の重症度に影響を与える可能性があることに注意してください。 </li><li>ハエはCO 2から移動性を回復するために5分を待ちます。それは5分で完全にCO 2の影響を除去するのに十分であるかどうかを知らないことに留意されたいです。 </li><li>ベルクロの内縁は、バイアル（ 図1B）の頂部と同じ高さになるまでバイアルに春の終わりを挿入します。春が平らに置かれた場合、バイアルの1インチ（2.5センチ）アイスバケットカバーの隆起領域をオーバーラップする必要があります。バイアルがmを再利用することができますいずれかの回。 </li><li>座った状態で、左手の親指と人差し指を使ってマジックテープ領域にバイアルを保持します。右手を使用して、ベンチトップに密な基板を保持します。あるいは、ベンチトップにタイトなボードを保持するために、C-クランプを使用しています。 </li><li>まっすぐ戻って所望の角度にバネを引き出します。春リリース。バネが完全に止まった状態にしてできるようにします。 </li><li>春からバイアルを取り外し、ハエが≥5分間回復することができます。別のストライキにハエを被写体やフライ食品でバイアルにハエを転送します。 注：種々のアッセイは、HIT装置からストライキの表現型効果を評価することができます。例えば、寿命への影響は、損傷後の時点で生存するハエの割合を分析することによって決定することができ、脳の形態への影響は、ヘッドの組織学的分析によって決定することができ、遺伝子発現に対する効果は、mRNAレベルの定量分析によって決定することができます10。 </li><li> Determinストライキに供されていない同一の対照ハエを処理することによるストライキによるものではない手続きの電子効果。アイスバケットカバーに対するバイアルの影響が大きな音を生成するので、保護を聞いて着用してください。 </li></ol>

<ol>
	<li>Harrison-Felix, C. L., Whiteneck, G. G., Jha, A., DeVivo, M. J., Hammond, F. M., Hart, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mortality+over+four+decades+after+traumatic+brain+injury+rehabilitation:+A+retrospective+cohort+study.">Mortality over four decades after traumatic brain injury rehabilitation: A retrospective cohort study.</a> Arch Phys Med Rehabil. 90, 1506-1513 (2009).</li><li>Coronado, V. G., 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=Surveillance+for+traumatic+brain+injury-related+deaths+&#8211;+United+States.">Surveillance for traumatic brain injury-related deaths &#8211; United States.</a> MMWR Surveill Summ. 60, 1-32 (1997).</li><li>Masel, B., DeWitt, D. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Traumatic+brain+injury:+A+disease+process,+not+an+event.">Traumatic brain injury: A disease process, not an event.</a> J. Neurotrauma. 27, 1529-1540 (2010).</li><li>Blennow, K., Hardy, J., Zetterberg, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+neuropathology+and+neurobiology+of+traumatic+brain+injury.">The neuropathology and neurobiology of traumatic brain injury.</a> Neuron. 76, 886-899 (2012).</li><li>Prins, M., Greco, T., Alexander, D., Giza, C. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+pathophysiology+of+traumatic+brain+injury+at+a+glance.">The pathophysiology of traumatic brain injury at a glance.</a> Disease Models Mech. 6, 1307-1315 (2013).</li><li>Menon, D. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Unique+challenges+in+clinical+trails+in+traumatic+brain+injury.">Unique challenges in clinical trails in traumatic brain injury.</a> Crit Care Med. 37, S129-S135 (2009).</li><li>Xiong, Y., Mahmood, A., Chopp, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+traumatic+brain+injury.">Animal models of traumatic brain injury.</a> Nature Rev Neurosci. 14, 128-142 (2013).</li><li>Balsiger, Z., Leudkte, J., Mawer, S., Willey, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> HIT device high speed analysis. , (2014).</li><li>Davceva, N., Janevska, V., Illevski, B., Petrushevska, G., Popeska, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+occurrence+of+acute+subdural+haematoma+and+diffuse+axonal+injury+as+two+typical+acceleration+injuries.">The occurrence of acute subdural haematoma and diffuse axonal injury as two typical acceleration injuries.</a> J Forensic Leg Med. 19, 480-484 (2012).</li><li>Katzenberger, R. J., Loewen, C. A., Wassarman, D. R., Petersen, A. J., Ganetzky, B., Wassarman , D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Drosophila.+model+of+closed+head+traumatic+brain+injury.">A Drosophila. model of closed head traumatic brain injury.</a> Proc Natl Acad Sci USA. 110, E4152-E4159 (2013).</li><li>Fang, Y., Bonini, N. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Axon+degeneration+and+regeneration:+insights+from+Drosophila+.models+of+nerve+injury.">Axon degeneration and regeneration: insights from Drosophila .models of nerve injury.</a> Annu Rev Cell Biol. 28, 575-597 (2012).</li><li>Babcock, D. T., Ganetzky, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+improved+method+for+accurate+and+rapid+measurement+of+flight+performance+in+Drosophila.">An improved method for accurate and rapid measurement of flight performance in Drosophila.</a> J Vis Exp. (84), e51223 (2014).</li><li>Tully, T., Preat, T., Boynton, S. C., Vecchio, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+dissection+of+consolidated+memory+in+Drosophila.">Genetic dissection of consolidated memory in Drosophila.</a> Cell. 79, 35-47 (1994).</li><li>Andretic, R., Shaw, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Essentials+of+sleep+recordings+in+Drosophila.:+moving+beyond+sleep+time.">Essentials of sleep recordings in Drosophila.: moving beyond sleep time.</a> Methods Enzymol. 393, 759-772 (2005).</li></ol>

我々は、開示する目的の競合がありません。

HIT装置方法は、それがTBI 11を貫通するのではなく、閉鎖性頭部を引き起こすという事実によってハエに外傷を負わせる他の方法と区別されます。また、HIT装置方法は、多くのハエでTBIを与えるために、より少ない時間、労力、およびスキルを取り、その方法は、大規模な遺伝子スクリーニングに、他の方法よりも適しています。最後に、HIT装置によって与え主要な傷害は、脳に限定?...

An author's name was corrected in the publication of <a href="http://www.jove.com/video/52905">A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila</a>. The second to last author's was spelled incorrectly. It has been updated from:
Barry Ganetky
to:
Barry Ganetzky

An author's name was corrected in the publication of <a href="http://www.jove.com/video/52905">A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila</a>.

Erratum: A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila

外傷性脳損傷（TBI）は、外部の機械的な力から脳への損傷として定義されます。このような鈍力と脳が頭蓋骨の内側に当たるせる慣性加速と減速力などの閉鎖性頭部力から最も一般的には、TBIの結果。米国では、50,000人がTBIから毎年死亡し、2.5から6500000個人は、物理的な認知および行動上の問題1,2を衰弱させるなど、TBIの結果、で生きていると推定されています。 TBIの結果は、脳への主要な機械的傷害にも時間3-5にわたって生じる脳への二次細胞および分子の怪我だけでなく、他の組織にだけではなくです。 TBIを診断し、治療するための方法の開発は、TBIは、複雑な疾患過程であるため、困難であることが証明されています。異種の二私の主な傷害、人間の生理、および環境要因の結果の可変性njuriesと病理。根本的な変動要因は、一次損傷の重症度、繰り返しの一次傷害の間の時間、および個々の年齢および遺伝子型が含まれます。各変動要因は、TBIの結果にどのように寄与するかを理解することは、TBI 6,7を診断し、治療するためのアプローチの開発に役立つ可能性があります。 ここでは、TBIの結果に変動要因の寄与を描写するために使用することができるキイロショウジョウバエ （ショウジョウバエ）で閉鎖性頭部TBIを負わする方法について説明します。手のひらに対して激しくフライ培養バイアルの側面を打つ最初の観察は、野生型は、TBIの可能性の高い結果を一時的に行動不能になるために飛ぶ引き起こさに方法がベースにしています。したがって、我々は手打つ作用から加速と減速力を再現するための高インパクトトラウマ（HIT）デバイスを構築しました。高速ムービーが示していることから、単一のストライキHITデバイスは、自分の頭と体8でバイアル壁を数回連絡するハエが発生します。ある程度までは、すべての連絡先は、ハエの脳が跳ね返ると転倒や車のクラッシュ9に人間に何が起こるかに似た頭部カプセル、に対して変形させる可能性があります。したがって、脳10で運動失調が続く一時的無能力、モビリティの緩やかな回復、頭の中で遺伝子発現の変化、および進行性の神経変性を含む、脳損傷と一致HITデバイスディスプレイの表現型で処理したハエ。このように、HITデバイスはハエのために開発された実験ツールと技術の巨大な武器庫を使用して、TBIを研究することが可能となります。

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Zinc plated compression spring</td>
			<td>The Hillman&#160; Group</td>
			<td>540189</td>
			<td>9 7/8 in (length), 15/16 in (outer diameter), 0.12 in (wire size)</td>
		</tr>
		<tr>
			<td>Wooden board</td>
			<td></td>
			<td></td>
			<td>9 in (length), 6.5 in (width), 0.75 in (height)</td>
		</tr>
		<tr>
			<td>Clamps</td>
			<td>Sigma Electrical Manufacturing Corporation</td>
			<td>49822</td>
			<td>3.10 in (length), 0.68 in (width), 1.11 in (height), EMT Two Hole Straps, click on type for 1 inch steel EMT conduit</td>
		</tr>
		<tr>
			<td>Loop half of self-adhesive velcro</td>
			<td></td>
			<td></td>
			<td>3 in (length), (3/4 in width)</td>
		</tr>
		<tr>
			<td>Polyurethane ice bucket cover</td>
			<td>Fisher Scientific</td>
			<td>02-591-45</td>
			<td>9 1/8 in (length), 9 1/8 in (width), 1 1/4 in (height)</td>
		</tr>
		<tr>
			<td>Plastic fly vials</td>
			<td>Applied Scientific</td>
			<td>AS-510</td>
			<td>3 11/16 in (height), 1 1/16 in (inner diameter), 1 1/8 in (outer diameter)</td>
		</tr>
		<tr>
			<td>Large cotton balls</td>
			<td>Fisher Scientific</td>
			<td>22-456-883</td>
			<td></td>
		</tr>
		<tr>
			<td>Paper protractor</td>
			<td></td>
			<td></td>
			<td>10 in (diameter)</td>
		</tr>
	</tbody>
</table>

a method to inflict closed head traumatic brain injury in drosophila

外傷性脳損傷（TBI）、物理的な認知、行動機能と死の障害を引き起こし、毎年何百万人もの人々に影響を与えます。 ショウジョウバエを用いた研究は、神経学的プロセスを理解する上で重要なブレークスルーを貢献しています。したがって、ヒトにおけるTBI病理の細胞および分子基盤を理解することを目標に、私たちはハエに閉鎖性頭部TBIを与えるためにハイインパクトトラウマ（HIT）装置を開発しました。ハエは、このような一時的な無能力および進行性神経変性などのヒトTBIと一致HITデバイス表示の表現型を行いました。 HITデバイスは、フライ脳への機械的損傷を引き起こし、バイアルの壁にハエを推進するために、スプリングベースのメ​​カニズムを使用しています。デバイスは、その操作は簡単で、迅速で、安価で構築することが容易であり、再現性のある結果を生成します。したがって、HIT装置は、既存の実験ツールとハエは基本対処するための技術と組み合わせることができTBIの診断と治療法の開発につながる可能性がTBIについての質問。具体的には、HITデバイスはTBI病状の遺伝的基礎を理解するために、大規模な遺伝子スクリーニングを行うことができます。

私たちは、ハエが一次損傷後まもなく死ぬ理由を理解することに興味を持っています。死を定量化するために、我々は、一次損傷の24時間以内に死亡したハエの割合である24時間で死亡率指数（MI 24）を 、決定しました。 HIT装置からストライキに供ハエは、ハエの餌とバイアル中で25℃でインキュベートし、そして死んだハエの数は、24時間後に計数しました。私たちは、1〜60分を試?...

Watch this Scientific Journal Video about A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila at JoVE.com

A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila

ここでは、 ショウジョウバエにおいて閉鎖性頭部外傷性脳損傷（TBI）を与えるための方法を説明します。この方法は、ハエのために利用可能な実験ツールと技術の広大な配列を使用して、TBI病理の根底にある細胞および分子メカニズムを調査するためのゲートウェイを提供します。

で閉鎖性頭部外傷性脳損傷を負わせる方法<em&gt;ショウジョウバエ</em

Traumatic brain injury (TBI) affects millions of people each year, causing impairment of physical, cognitive, and behavioral functions and death. Studies ...

a-method-to-inflict-closed-head-traumatic-brain-injury-in-drosophila

Research

JoVE Journal

Neuroscience

A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila

11.4K ビュー.  University of Wisconsin-Madison. ここでは、 ショウジョウバエにおいて閉鎖性頭部外傷性脳損傷（TBI）を与えるための方法を説明します。この方法は、ハエのために利用可能な実験ツールと技術の広大な配列を使用して、TBI病理の根底にある細胞および分子メカニズムを調査するためのゲートウェイを提供します。 

ビデオ: A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila

Lateral Fluid Percussion: Model of Traumatic Brain Injury in Mice

Traumatic brain injury (TBI) research has attained renewed momentum due to the increasing awareness of head injuries, which result in morbidity and mortality. Based on the nature of primary injury following TBI, complex and heterogeneous secondary consequences result, which are followed by regenerative processes 1,2. Primary injury can be induced by a direct contusion to the brain from skull fracture or from shearing and stretching of tissue causing displacement of brain due to movement 3,4. The resulting hematomas and lacerations cause a vascular response 3,5, and the morphological and functional damage of the white matter leads to diffuse axonal injury 6-8. Additional secondary changes commonly seen in the brain are edema and increased intracranial pressure 9. Following TBI there are microscopic alterations in biochemical and physiological pathways involving the release of excitotoxic neurotransmitters, immune mediators and oxygen radicals 10-12, which ultimately result in long-term neurological disabilities 13,14. Thus choosing appropriate animal models of TBI that present similar cellular and molecular events in human and rodent TBI is critical for studying the mechanisms underlying injury and repair.
Various experimental models of TBI have been developed to reproduce aspects of TBI observed in humans, among them three specific models are widely adapted for rodents: fluid percussion, cortical impact and weight drop/impact acceleration 1. The fluid percussion device produces an injury through a craniectomy by applying a brief fluid pressure pulse on to the intact dura. The pulse is created by a pendulum striking the piston of a reservoir of fluid. The percussion produces brief displacement and deformation of neural tissue 1,15. Conversely, cortical impact injury delivers mechanical energy to the intact dura via a rigid impactor under pneumatic pressure 16,17. The weight drop/impact model is characterized by the fall of a rod with a specific mass on the closed skull 18. Among the TBI models, LFP is the most established and commonly used model to evaluate mixed focal and diffuse brain injury 19. It is reproducible and is standardized to allow for the manipulation of injury parameters. LFP recapitulates injuries observed in humans, thus rendering it clinically relevant, and allows for exploration of novel therapeutics for clinical translation 20.
We describe the detailed protocol to perform LFP procedure in mice. The injury inflicted is mild to moderate, with brain regions such as cortex, hippocampus and corpus callosum being most vulnerable. Hippocampal and motor learning tasks are explored following LFP.

Lateral fluid percussion (LFP), an established model of traumatic brain injury in mice, is demonstrated. LFP fulfills three major criteria for animal models: validity, reliability and clinical relevance. The procedure, consisting of surgical craniotomy, fixation of hub followed by induction of injury, resulting in focal and diffuse injuries, is described.

横方向の流体のパーカッション：マウスにおける外傷性脳損傷のモデル

Traumatic brain injury (TBI) research has attained renewed momentum due to the increasing awareness of head injuries, which result in morbidity and ...

神経科学

Investigations on Alterations of Hippocampal Circuit Function Following Mild Traumatic Brain Injury

Traumatic Brain Injury (TBI) afflicts more than 1.7 million people in the United States each year and even mild TBI can lead to persistent neurological impairments 1. Two pervasive and disabling symptoms experienced by TBI survivors, memory deficits and a reduction in seizure threshold, are thought to be mediated by TBI-induced hippocampal dysfunction 2,3. In order to demonstrate how altered hippocampal circuit function adversely affects behavior after TBI in mice, we employ lateral fluid percussion injury, a commonly used animal model of TBI that recreates many features of human TBI including neuronal cell loss, gliosis, and ionic perturbation 4-6.
	Here we demonstrate a combinatorial method for investigating TBI-induced hippocampal dysfunction. Our approach incorporates multiple ex vivo physiological techniques together with animal behavior and biochemical analysis, in order to analyze post-TBI changes in the hippocampus. We begin with the experimental injury paradigm along with behavioral analysis to assess cognitive disability following TBI. Next, we feature three distinct ex vivo recording techniques: extracellular field potential recording, visualized whole-cell patch-clamping, and voltage sensitive dye recording. Finally, we demonstrate a method for regionally dissecting subregions of the hippocampus that can be useful for detailed analysis of neurochemical and metabolic alterations post-TBI.
	These methods have been used to examine the alterations in hippocampal circuitry following TBI and to probe the opposing changes in network circuit function that occur in the dentate gyrus and CA1 subregions of the hippocampus (see Figure 1). The ability to analyze the post-TBI changes in each subregion is essential to understanding the underlying mechanisms contributing to TBI-induced behavioral and cognitive deficits. 
	The multi-faceted system outlined here allows investigators to push past characterization of phenomenology induced by a disease state (in this case TBI) and determine the mechanisms responsible for the observed pathology associated with TBI.

A multi-faceted approach to investigating functional changes to hippocampal circuitry is explained. Electrophysiological techniques are described along with the injury protocol, behavioral testing and regional dissection method. The combination of these techniques can be applied in similar fashion for other brain regions and scientific questions.

軽度の外傷性脳損傷後の海馬回路機能の改変に関する調査

Traumatic Brain Injury (TBI) afflicts more than 1.7 million people in the United States each year and even mild TBI can lead to persistent neurological ...

A Novel Method to Model Chronic Traumatic Encephalopathy in Drosophila

Chronic Traumatic Encephalopathy (CTE) is an established neurodegenerative disease that is closely associated with exposure to repetitive mild Traumatic Brain Injury (mTBI). The mechanisms responsible for its complex pathological changes remain largely elusive, despite a recent consensus to define the neuropathological criteria. Here, we describe a novel method to develop a model of CTE in Drosophila melanogaster&#160;(Drosophila&#160;)&#160;in an attempt to identify the key genes and pathways that lead to the characteristic hyperphosphorylated tau accumulation and neuronal death in the brain. Adjustable-strength impacts to inflict mild closed injury are delivered directly to the fly head, subjecting the head to rapid acceleration and deceleration. Our method eliminates the potential problems inherent with other Drosophila mTBI models (e.g.,animal death might be induced by damage to other parts of the body or to internal organs). The less labor- and cost-intensive animal care, short life span, and extensive genetic tools make the fruit fly ideal to study CTE pathogenesis and make it possible to perform large-scale, genome-wide forward genetic and pharmacological screens. We anticipate that the ongoing characterization of the model will generate important mechanistic insights on disease prevention and therapeutic approaches.

A Novel Method to Model Chronic Traumatic Encephalopathy in Drosophila

Here, we describe a new approach to inflict closed-head traumatic brain injury in Drosophila melanogaster. Our method has the advantage of directly delivering repetitive impacts with adjustable strength to the head alone. Further exploration of the invertebrate system will help to illuminate the pathogenesis of chronic traumatic encephalopathy.

慢性外傷性脳症モデルの新しい方法<em&gt;ショウジョウバエ</em

Chronic Traumatic Encephalopathy (CTE) is an established neurodegenerative disease that is closely associated with exposure to repetitive mild Traumatic ...

Stereotactic Atlas-Guided Laser Capture Microdissection of Brain Regions Affected by Traumatic Injury

The ability to isolate specific brain regions of interest can be impeded in tissue disassociation techniques that do not preserve their spatial distribution. Such techniques also potentially skew gene expression analysis because the process itself can alter expression patterns in individual cells. Here we describe a laser capture microdissection (LCM) method to selectively collect specific brain regions affected by traumatic brain injury (TBI) by using a modified Nissl (cresyl violet) staining protocol and the guidance of a rat brain atlas. LCM provides access to brain regions in their native positions and the ability to use anatomical landmarks for identification of each specific region. To this end, LCM has been used previously to examine brain region specific gene expression in TBI. This protocol allows examination of TBI-induced alterations in gene and microRNA expression in distinct brain areas within the same animal. The principles of this protocol can be amended and applied to a wide range of studies examining genomic expression in other disease and/or animal models.

We describe the use of laser capture microdissection to obtain samples of distinct cell populations from different brain regions for gene and microRNA analysis. This technique allows the study of differential effects of traumatic brain injury in specific regions of the rat brain.

外傷によって影響を受ける脳のレーザーキャプチャーマイクロダイ セクション定位アトラス誘導レーザー

The ability to isolate specific brain regions of interest can be impeded in tissue disassociation techniques that do not preserve their spatial ...

A Novel In Vitro Model of Blast Traumatic Brain Injury

Traumatic brain injury is a leading cause of death and disability in military and civilian populations. Blast traumatic brain injury results from the detonation of explosive devices, however, the mechanisms that underlie the brain damage resulting from blast overpressure exposure are not entirely understood and are believed to be unique to this type of brain injury. Preclinical models are crucial tools that contribute to better understand blast-induced brain injury. A novel in vitro blast TBI model was developed using an open-ended shock tube to simulate real-life open-field blast waves modelled by the Friedlander waveform. C57BL/6N mouse organotypic hippocampal slice cultures were exposed to single shock waves and the development of injury was characterized up to 72 h using propidium iodide, a well-established fluorescent marker of cell damage that only penetrates cells with compromised cellular membranes. Propidium iodide fluorescence was significantly higher in the slices exposed to a blast wave when compared with sham slices throughout the duration of the protocol. The brain tissue injury is very reproducible and proportional to the peak overpressure of the shock wave applied.

A Novel In Vitro Model of Blast Traumatic Brain Injury

This paper describes a novel model of primary blast traumatic brain injury. A compressed-air driven shock tube is used to expose in vitro mouse hippocampal slice cultures to a single shock wave. This is a simple and rapid protocol generating a reproducible brain tissue injury with a high throughput.

爆発外傷性脳損傷の新規In Vitroモデル

Traumatic brain injury is a leading cause of death and disability in military and civilian populations. Blast traumatic brain injury results from the ...

Effects of Blast-induced Neurotrauma on Pressurized Rodent Middle Cerebral Arteries

Though there have been studies on the histopathological and behavioral effects of blast exposure, fewer have been dedicated to blast&#39;s cerebral vascular effects. Impact (i.e., non-blast) traumatic brain injury (TBI) is known to decrease pressure autoregulation in the cerebral vasculature in both humans and experimental animals. The hypothesis that blast-induced traumatic brain injury (bTBI), like impact TBI, results in impaired cerebral vascular reactivity was tested by measuring myogenic dilatory responses to reduced intravascular pressure in rodent middle cerebral arterial (MCA) segments from rats subjected to mild bTBI using an Advanced Blast Simulator (ABS) shock tube. Adult, male Sprague-Dawley rats were anesthetized, intubated, ventilated and prepared for Sham bTBI (identical manipulation and anesthesia except for blast injury) or mild bTBI. Rats were randomly assigned to receive Sham bTBI or mild bTBI followed by sacrifice 30 or 60 min post-injury. Immediately after bTBI, righting reflex (RR) suppression times were assessed, euthanasia at the time points post-injury was completed, the brain was harvested and the individual MCA segments were collected, mounted and pressurized. As the intraluminal pressure perfused through the arterial segments was reduced in 20 mmHg increments from 100 to 20 mmHg, MCA diameters were measured and recorded. With decreasing intraluminal pressure, MCA diameters steadily increased significantly above baseline in the Sham bTBI groups while MCA dilator responses were significantly reduced (p &#60; 0.05) in both bTBI groups as evidenced by the impaired, smaller MCA diameters recorded for the bTBI groups. In addition, RR suppression in the bTBI groups was significantly (p &#60; 0.05) higher than in the Sham bTBI groups. MCA&#39;s collected from the Sham bTBI groups exhibited typical vasodilatory properties to decreases in intraluminal pressure while MCA&#39;s collected following bTBI exhibited significantly impaired myogenic vasodilatory responses to reduced pressure that persisted for at least 60 min after bTBI.

Here, we present a protocol to describe methods for ex vivo vascular reactivity determination following a primary blast traumatic brain injury (bTBI) using isolated, pressurized, rodent middle cerebral arterial (MCA) segments. bTBI induction is accomplished using a shock tube, also known as an Advanced Blast Simulator (ABS) device.

加圧齧歯動物に及ぼす影響の爆発による頭部外傷の中間の大脳動脈

Though there have been studies on the histopathological and behavioral effects of blast exposure, fewer have been dedicated to blast&#39;s cerebral ...

Investigating Alterations in Caecum Microbiota After Traumatic Brain Injury in Mice

Increasing evidence shows that the microbiota-gut-brain axis plays an important role in the pathogenesis of brain diseases. Several studies also demonstrate that traumatic brain injuries cause changes to the gut microbiota. However, mechanisms underlying the bidirectional regulation of the brain-gut axis remain unknown. Currently, few models exist for studying the changes in gut microbiota after traumatic brain injury. Therefore, the presented study combines protocols for inducing traumatic brain injury using a lateral fluid percussion device and analysis of caecum samples following injury for investigating alterations in the gut microbiome. Alterations of the gut microbiota composition after traumatic brain injury are determined using 16S-rDNA sequencing. This protocol provides an effective method for studying the relationships between enteric microorganisms and traumatic brain injury.

Presented here is a protocol to induce diffuse traumatic brain injury using a lateral fluid percussion device followed by the collection of the caecum content for gut microbiome analysis.

マウスの外傷性脳損傷後のカエカム微生物叢の変化を調べている

Increasing evidence shows that the microbiota-gut-brain axis plays an important role in the pathogenesis of brain diseases. Several studies also ...

Low-intensity Blast Wave Model for Preclinical Assessment of Closed-head Mild Traumatic Brain Injury in Rodents

Traumatic brain injury (TBI) is a large-scale public health problem. Mild TBI is the most prevalent form of neurotrauma and accounts for a large number of medical visits in the United States. There are currently no FDA-approved treatments available for TBI. The increased incidence of military-related, blast-induced TBI further accentuates the urgent need for effective TBI treatments. Therefore, new preclinical TBI animal models that recapitulate aspects of human blast-related TBI will greatly advance the research efforts into the neurobiological and pathophysiological processes underlying mild to moderate TBI as well as the development of novel therapeutic strategies for TBI.
Here we present a reliable, reproducible model for the investigation of the molecular, cellular, and behavioral effects of mild to moderate blast-induced TBI. We describe a step-by-step protocol for closed-head, blast-induced mild TBI in rodents using a bench-top setup consisting of a gas-driven shock tube equipped with piezoelectric pressure sensors to ensure consistent test conditions. The benefits of the setup that we have established are its relative low-cost, ease of installation, ease of use and high-throughput capacity. Further advantages of this non-invasive TBI model include the scalability of the blast peak overpressure and the generation of controlled reproducible outcomes. The reproducibility and relevance of this TBI model has been evaluated in a number of downstream applications, including neurobiological, neuropathological, neurophysiological and behavioral analyses, supporting the use of this model for the characterization of processes underlying the etiology of mild to moderate TBI.

We present here a protocol of a blast wave model for rodents to investigate neurobiological and pathophysiological effects of mild to moderate traumatic brain injury. We established a gas-driven, bench-top setup equipped with pressure sensors allowing for reliable and reproducible generation of blast-induced mild to moderate traumatic brain injury.

げっ歯類における閉鎖性軽度外傷性脳損傷の前臨床評価のための低強度ブラスト波モデル

Traumatic brain injury (TBI) is a large-scale public health problem. Mild TBI is the most prevalent form of neurotrauma and accounts for a large number of ...

Systems Analysis of the Neuroinflammatory and Hemodynamic Response to Traumatic Brain Injury

Mild traumatic brain injuries (mTBIs) are a significant public health problem. Repeated exposure to mTBI can lead to cumulative, long-lasting functional deficits. Numerous studies by our group and others have shown that mTBI stimulates cytokine expression and activates microglia, decreases cerebral blood flow and metabolism, and impairs cerebrovascular reactivity. Moreover, several works have reported an association between derangements in these neuroinflammatory and hemodynamic markers and cognitive impairments. Herein we detail methods to characterize the neuroinflammatory and hemodynamic tissue response to mTBI in mice. Specifically, we describe how to perform a weight-drop model of mTBI, how to longitudinally measure cerebral blood flow using a non-invasive optical technique called diffuse correlation spectroscopy, and how to perform a Luminex multiplexed immunoassay on brain tissue samples to quantify cytokines and immunomodulatory phospho-proteins (e.g., within the MAPK and NF&#954;B pathways) that respond to and regulate activity of microglia and other neural immune cells. Finally, we detail how to integrate these data using a multivariate systems analysis approach to understand the relationships between all of these variables. Understanding the relationships between these physiologic and molecular variables will ultimately enable us to identify mechanisms responsible for mTBI.

This protocol presents methods to characterize the neuroinflammatory and hemodynamic response to mild traumatic brain injury and to integrate these data as part of a multivariate systems analysis using partial least squares regression.

外傷性脳損傷に対する神経炎症および血行動態応答のシステム解析

Mild traumatic brain injuries (mTBIs) are a significant public health problem. Repeated exposure to mTBI can lead to cumulative, long-lasting functional ...

Electromagnetic Controlled Closed-Head Model of Mild Traumatic Brain Injury in Mice

Highly reproducible animal models of traumatic brain injury (TBI), with well-defined pathologies, are needed for testing therapeutic interventions and understanding the mechanisms of how a TBI alters brain function. The availability of multiple animal models of TBI is necessary to model the different aspects and severities of TBI seen in people. This manuscript describes the use of a midline closed head injury (CHI) to develop a mouse model of mild TBI. The model is considered mild because it does not produce structural brain lesions based on neuroimaging or gross neuronal loss. However, a single impact creates enough pathology that cognitive impairment is measurable at least 1 month after injury. A step-by-step protocol to induce a CHI in mice using a stereotaxically guided electromagnetic impactor is defined in the paper. The benefits of the mild midline CHI model include the reproducibility of the injury-induced changes with low mortality. The model has been temporally characterized up to 1 year after the injury for neuroimaging, neurochemical, neuropathological, and behavioral changes. The model is complementary to open skull models of controlled cortical impact using the same impactor device. Thus, labs can model both mild diffuse TBI and focal moderate-to-severe TBI with the same impactor.

The protocol describes mild traumatic brain injury in a mouse model. In particular, a step-by-step protocol to induce a mild midline closed head injury and the characterization of the animal model is fully explained.

マウスにおける軽度外傷性脳損傷の電磁制御クローズドヘッドモデル

Highly reproducible animal models of traumatic brain injury (TBI), with well-defined pathologies, are needed for testing therapeutic interventions and ...