Name: lateral chronic cranial window preparation enables in vivo observation following distal middle cerebral artery occlusion in mice
Uploaded: 2016-12-29T13:00:00
Description: Watch this Scientific Journal Video about Lateral Chronic Cranial Window Preparation Enables In Vivo Observation Following Distal Middle Cerebral Artery Occlusion in Mice at JoVE.com

VP is a participant in the Charit&#233; Clinical Scientist Program, funded by the Charit&#233; - Universit&#228;tsmedizin Berlin and the Berlin Institute of Health. TB is an SNSF PostDoc Mobility fellow. The authors receive grant support from EinsteinStiftung/A-2012-153 to PV.

倫理の声明：動物を対象とした実験は、該当する場合、Landesamt fuerお大事にウントSoziales、ベルリン、ドイツ（G0298 / 13）とARRIVE基準によって定められたガイドラインおよび規則に従って行いました。 12週齢の雄のC57BL / 6Jマウスに本研究では、10-に使用しました。 1.横慢性頭蓋窓の準備<ol><li>皮下ケタミンの注射（90 mgの/ kg）およびキシラジン（10mg / kg）で麻酔を行いま?...

<ol>
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(61), (2012).</li><li>Holtmaat, A., Bonhoeffer, T., 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=Long-term,+high-resolution+imaging+in+the+mouse+neocortex+through+a+chronic+cranial+window.">Long-term, high-resolution imaging in the mouse neocortex through a chronic cranial window.</a> Nat Protoc. 4 (8), 1128-1144 (2009).</li><li>Iadecola, C., Dirnagl, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+microcircualtion--fantastic+voyage:+introduction.">The microcircualtion--fantastic voyage: introduction.</a> Stroke. 44 (6), 83 (2013).</li><li>Blinder, P., Tsai, P. S., Kaufhold, J. P., Knutsen, P. 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Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+intravital+thinned+skull+and+cranial+window+approaches+to+study+CNS+immunobiology+in+the+mouse+cortex.">Comparison of intravital thinned skull and cranial window approaches to study CNS immunobiology in the mouse cortex.</a> Intravital. 3 (2), (2014).</li><li>Balkaya, M., 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=Assessing+post-stroke+behavior+in+mouse+models+of+focal+ischemia.">Assessing post-stroke behavior in mouse models of focal ischemia.</a> J Cereb Blood Flow Metab. 33 (3), 330-338 (2013).</li><li>Balkaya, M., Kr&#246;ber, J., Gertz, K., Peruzzaro, S., Endres, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+long-term+functional+outcome+in+a+murine+model+of+mild+brain+ischemia.">Characterization of long-term functional outcome in a murine model of mild brain ischemia.</a> J Neurosci Methods. 213 (2), 179-187 (2013).</li><li>Freret, T., Bouet, V., 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=Behavioral+deficits+after+distal+focal+cerebral+ischemia+in+mice:+Usefulness+of+adhesive+removal+test.">Behavioral deficits after distal focal cerebral ischemia in mice: Usefulness of adhesive removal test.</a> Beh Neurosci. 123 (1), 224-230 (2009).</li><li>Liu, S., Zhen, G., Meloni, B. P., Campbell, K., Winn, H. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=RODENT+STROKE+MODEL+GUIDELINES+FOR+PRECLINICAL+STROKE+TRIALS+(1ST+EDITION).">RODENT STROKE MODEL GUIDELINES FOR PRECLINICAL STROKE TRIALS (1ST EDITION).</a> J Exp Stroke Trans Med. 2 (2), 2-27 (2009).</li><li>Florian, B., Vintilescu, R., 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=Long-term+hypothermia+reduces+infarct+volume+in+aged+rats+after+focal+ischemia.">Long-term hypothermia reduces infarct volume in aged rats after focal ischemia.</a> Neurosci Lett. 438 (2), 180-185 (2008).</li><li>Noor, R., Wang, C. X., Shuaib, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+hyperthermia+on+infarct+volume+in+focal+embolic+model+of+cerebral+ischemia+in+rats.">Effects of hyperthermia on infarct volume in focal embolic model of cerebral ischemia in rats.</a> Neurosci Lett. 349 (2), 130-132 (2003).</li><li>Barber, P. A., Hoyte, L., Colbourne, F., Buchan, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Temperature-regulated+model+of+focal+ischemia+in+the+mouse:+a+study+with+histopathological+and+behavioral+outcomes.">Temperature-regulated model of focal ischemia in the mouse: a study with histopathological and behavioral outcomes.</a> Stroke. 35 (7), 1720-1725 (2004).</li><li>Shin, H. 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ストロークが1世界中の長期障害と死の主な原因の一つです。急性期治療を超えて、脳卒中後の回復を促進し、強化するための新しいアプローチやメカニズムを調査すると、プライム医療目標7のまま。実験ストローク研究者が頻繁に局所脳虚血の齧歯類モデルを採用しています。実際には、一時的または恒久的にMCAoを誘導するモデルは、患者4における局所脳虚血の最...

Stroke is among the principal causes of long-term disability and death worldwide1, coming second after coronary heart disease. In addition, stroke is the primary cause of long-term disability, underscoring its tremendous socioeconomic impact6-8. Beyond acute treatment, investigating new approaches and mechanisms to accelerate and enhance recovery after stroke remains a prime medical goal7. 
In the last few decades, data from experimental stroke research has contributed substantially to understanding the complex pathophysiological cascades triggered by ischemia9,10. Excitotoxicity, apoptosis, peri-infarct depolarization, and inflammation have been identified as the most relevant mediators of cell death following focal cerebral ischemia. Moreover, using animal models of cerebral ischemia, important concepts, diagnostic modalities, and therapeutic approaches have been developed and validated (e.g., "penumbra" and thrombolysis)11.
The availability of experimental stroke models, combined with non-invasive imaging modalities (e.g., magnetic resonance imaging (MRI), computed tomography, or laser speckle contrast analysis), enables the researcher to investigate hyperacute and chronic pathophysiological changes induced by the ischemic insult in a longitudinal manner12. Along with studying the spatiotemporal profile of the evolving lesion, changes resembling neuronal plasticity can be investigated and correlated to functional outcomes and histological findings. Within the last few years, further methodological advances have been made using the combination of cerebral ischemia models and in vivo microscopy via cranial windows13. These new techniques allow investigators to analyze the neurovascular unit at the cellular and molecular level, with great analytic power in the acute, subacute, and chronic phases following focal cerebral ischemia14. Moreover, in vivo microscopy imaging of microcirculatory dynamics has revealed novel aspects of cerebral microvasculature function and angioarchitecture, with significant pathophysiological relevance15-17. 
In this protocol, we present how to perform a chronic CW preparation lateral to the sagittal sinus and how to surgically induce a distal stroke underneath the window. This mouse model can be applied to sequential imaging of acute, subacute, and chronic changes following focal cerebral ischemia via epi-illuminating, confocal laser scanning, and two-photon intravital microscopy.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Binocular surgical microscope</td>
			<td>Zeiss</td>
			<td>Stemi 2000 C</td>
			<td></td>
		</tr>
		<tr>
			<td>Light source for microscope</td>
			<td>Zeiss</td>
			<td>CL 6000 LED</td>
			<td></td>
		</tr>
		<tr>
			<td>Heating pad with rectal probe</td>
			<td>FST</td>
			<td>21061-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Stereotactic frame</td>
			<td>Kopf</td>
			<td>Model 930</td>
			<td></td>
		</tr>
		<tr>
			<td>Anaethesia system for isoflurane</td>
			<td>Draeger</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Abott</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Dumont forceps #5</td>
			<td>FST</td>
			<td>11251-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Dumont forceps #7</td>
			<td>FST</td>
			<td>11271-30</td>
			<td></td>
		</tr>
		<tr>
			<td>Bipolar Forceps</td>
			<td>Erbe</td>
			<td>20195-501</td>
			<td></td>
		</tr>
		<tr>
			<td>Bipolar Forceps&#160;</td>
			<td colspan="2">Erbe&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; 20195-022</td>
			<td></td>
		</tr>
		<tr>
			<td>Microdrill</td>
			<td colspan="2">FST&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; 18000-17&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Needle holder</td>
			<td>FST</td>
			<td>12010-14</td>
			<td></td>
		</tr>
		<tr>
			<td>5-0 silk suture</td>
			<td>Feuerstein, Suprama</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>7-0 silk suture</td>
			<td>Feuerstein,Suprama</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>8-0 silk suture</td>
			<td>Feuerstein, Suprama</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Veterinary Recovery Chamber</td>
			<td>Peco Services</td>
			<td>V1200</td>
			<td></td>
		</tr>
	</tbody>
</table>

lateral chronic cranial window preparation enables in vivo observation following distal middle cerebral artery occlusion in mice

局所脳虚血（ すなわち、虚血性脳卒中）は、神経機能の深刻な損失に、その結果、運動および認知障害のホストにつながる、主要な脳損傷を引き起こす可能性があります。ストロークが1世界中の長期障害と死の主な原因の一つであるとして、その高い有病率は、深刻な健康負担をもたらします。神経機能の回復は、ほとんどの場合、唯一の部分です。これまでに、治療オプションが原因で血栓溶解2,3ための狭い時間ウィンドウに、特に、非常に制限されます。脳卒中からの回復を加速するための方法を決定することは、プライム医療目標です。しかしながら、これは、回復プロセス中に不十分な機構的洞察により妨げられてきました。実験ストローク研究者が頻繁に局所脳虚血の齧歯類モデルを採用しています。急性期を越えて、脳卒中の研究は、脳虚血次亜急性および慢性期にますます集中しています。ほとんどのストロークの研究者は、永続的またはトラン適用しますマウスまたはラットにおけるMCAのトランジェント閉塞。患者では、MCAの閉塞は、虚血性脳卒中4の最も頻繁な原因の一つです。フィラメントモデルを用いて、MCAの近位閉塞のほか、遠位MCAの外科的閉塞は実験的ストローク研究5の中で最も頻繁に使用されるモデルは、おそらくです。 MCAブランチ（lenticulo-線条動脈の分岐まで）遠位の閉塞は、通常、線条体を惜しみ、主に新皮質に影響を与えます。血管閉塞は、永久的または一時的であることができます。長期転帰に対する病変体積と非常に低い死亡率の高い再現性は、このモデルの主な利点です。ここでは、矢状静脈洞に慢性頭蓋窓（CW）準備横を実行するために、その後どのように開頭術アプローチを使用して、ウィンドウの下に遠位脳卒中を引き起こす外科的方法を示しています。このアプローチは、ビアの虚血後の急性および慢性の変化の連続画像形成のために適用することができます落射照明、共焦点レーザー走査、及び二光子生体顕微鏡。

タイムライン及び代表的な結果を図2および図3に示されています。上矢状静脈洞（ 図2 B、C、D）経験豊富な外科医によって行われる非常に低い死亡率および罹患率率の結果に小さな頭蓋窓の横で頭蓋窓の準備、。 10匹の動物の全てが生存し、そしてすべての慢性CWも28日目、手術後、高品質の画像化に使用することができます。創?...

Watch this Scientific Journal Video about Lateral Chronic Cranial Window Preparation Enables In Vivo Observation Following Distal Middle Cerebral Artery Occlusion in Mice at JoVE.com

Lateral Chronic Cranial Window Preparation Enables In Vivo Observation Following Distal Middle Cerebral Artery Occlusion in Mice

遠位中大脳動脈の分岐（MCAO）の外科的閉塞は実験的ストローク研究で頻繁に使用されるモデルです。この原稿は、マウスの縦生体顕微鏡検査のための機会を提供しています横頭蓋窓の挿入と組み合わせて永久にMCAoの基本的な手法について説明します。

横慢性頭蓋窓の準備を有効<em&gt;インビボ</em&gt;マウスの遠位中大脳動脈閉塞後に観測

Focal cerebral ischemia (i.e., ischemic stroke) may cause major brain injury, leading to a severe loss of neuronal function and consequently to a host of ...

Research

JoVE Journal

Neuroscience

Lateral Chronic Cranial Window Preparation Enables In Vivo Observation Following Distal Middle Cerebral Artery Occlusion in Mice

Focal Cerebral Ischemia Model by Endovascular Suture Occlusion of the Middle Cerebral Artery in the Rat

ラット中大脳動脈の血管縫合閉塞による局所脳虚血モデル

Stroke is the leading cause of disability and the third leading cause of death in adults worldwide1. In human stroke, there exists a highly variable ...

Dual Electrophysiological Recordings of Synaptically-evoked Astroglial and Neuronal Responses in Acute Hippocampal Slices

急性海馬スライスにおけるシナプス誘発アストログリアとニューロン活動の二重の電気生理学的記録

Astrocytes form together with neurons tripartite synapses, where they integrate and modulate neuronal activity. Indeed, astrocytes sense neuronal inputs ...

Piezo High Accuracy Surgical Osteal Removal (PHASOR): A Technique for Improved Cranial Window Surgery in Mice

Piezo High Accuracy Surgical Osteal Removal PHASOR: A Technique for Improved Cranial Window Surgery in Mice

ピエゾ高精度手術小片除去 (フェーザ): マウスの改良された頭蓋ウィンドウ術のテクニック

Multiphoton microscopy has been widely adapted for imaging neurons in vivo. Repeated imaging requires implantation of a cranial window or repeated ...

Modeling Stroke in Mice: Transient Middle Cerebral Artery Occlusion via the External Carotid Artery

マウスにおける脳卒中のモデリング:外頸動脈を介した過渡中脳動脈閉塞

Stroke is the third most common cause of mortality and the leading cause of acquired adult disability in developed countries. To date, therapeutic options ...

Implantation of a Cranial Window for Repeated In Vivo Imaging in Awake Mice

Implantation of a Cranial Window for Repeated In Vivo Imaging in Awake Mice

覚醒マウスにおける 反復生体内 イメージングのための頭蓋窓の移植

To fully understand the cellular physiology of neurons and glia in behaving animals, it is necessary to visualize their morphology and record their ...

Construction and Implementation of Carbon Fiber Microelectrode Arrays for Chronic and Acute In Vivo Recordings

Construction and Implementation of Carbon Fiber Microelectrode Arrays for Chronic and Acute In Vivo Recordings

慢性および急性イン ビボ 記録用炭素繊維微小電極アレイの構築と実装

Multichannel electrode arrays offer insight into the working brain and serve to elucidate neural processes at the single-cell and circuit levels. ...

A Model for Encephalomyosynangiosis Treatment after Middle Cerebral Artery Occlusion-Induced Stroke in Mice

マウスにおける中大脳動脈閉塞性脳卒中後の脳筋シナジア症治療モデル

There is no effective treatment available for most patients suffering with ischemic stroke, making development of novel therapeutics imperative. The ...

In Vivo Wide-Field and Two-Photon Calcium Imaging from a Mouse Using a Large Cranial Window

In Vivo Wide-Field and Two-Photon Calcium Imaging from a Mouse Using a Large Cranial Window

インビボ 大きな頭蓋窓を用いたマウスからの広視野および2光子カルシウムイメージング

Wide-field calcium imaging from the mouse&#39;s neocortex allows one to observe cortex-wide neural activity related to various brain functions. On the ...

Assessment of Thermal Damage from Robot-Drilled Craniotomy for Cranial Window Surgery in Mice

マウスの頭蓋窓手術のためのロボットドリル開頭術による熱損傷の評価

Cranial window surgery allows for the imaging of brain tissue in live mice with the use of multiphoton or other intravital imaging techniques. However, ...

Establishing a Transcranial Middle Cerebral Artery Occlusion Mice Model

A Modified Transcranial Middle Cerebral Artery Occlusion Model to Study Stroke Outcomes in Aged Mice

Innovative Techniques and Future Directions in Stroke Research (Video) | JoVE

高齢マウスの脳卒中転帰を研究するための修正経頭蓋中大脳動脈閉塞モデル

In experimental stroke research, middle cerebral artery occlusion (MCAO) with an intraluminal filament is widely used to model ischemic stroke in mice. ...