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.

윤리 선언문 : 동물 과목을 포함하는 실험은 적용 가능한 Landesamt FUER 아퍼 싶게 Soziales, 베를린, 독일 (G0298 / 13) 및 도착 도착 기준에 명시된 지침과 규정에 따라 수행되었다. 12 주령의 수컷 C57BL / 6J 마우스에 대한 연구, 10을 사용 하였다. 1. 측면 만성 두개골 창 준비 <ol><li> 케타민 (90 ㎎ / ㎏)과 자일 라진 (/ kg 10 mg)의 피하 주사로 마취를 수행합니다. 통증 자극에 적절한 진정 ...

<ol>
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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. 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스트로크 1 전 세계적으로 장기 장애와 사망의 주요 원인 중 하나입니다. 급성 치료, 가속 및 뇌졸중 후 회복을 향상시키기 위해 새로운 접근 방식과 메커니즘을 조사 외에도 주요 의료 목표 7 남아있다. 실험 행정 연구원은 자주 국소 뇌허혈의 설치류 모델을 사용한다. 사실, 일시적 또는 영구적 유도 MCAO 모델은 환자 4 국소 뇌허혈의 가장 흔한 유형 중 하나를 모방. MCA 폐?...

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

초점 대뇌 허혈 (즉, 허혈성 뇌졸중)은 신경 기능의 심각한 손실을 결과적으로 모터 및인지 장애의 호스트로 이어지는 주요 뇌 손상의 원인이 될 수 있습니다. 스트로크 1 전 세계적으로 장기 장애와 사망의 주요 원인 중 하나입니다으로 높은 유병률은, 심각한 건강 부담을 포즈. 신경 기능의 회복은, 대부분의 경우, 단지 부분이다. 지금까지 치료 옵션은 매우 인해 혈전 용해 2,3의 좁은 시간 창에 특히 제한됩니다. 뇌졸중에서 회복을 가속화하는 방법을 결정하는 주요 의료 목표 남아있다; 그러나, 이는 복구 프로세스에 충분한 기계적 통계에 의해 방해되어왔다. 실험 행정 연구원은 자주 국소 뇌허혈의 설치류 모델을 사용한다. 급성기를 넘어, 뇌졸중 연구는 점점 뇌허혈 다음 서브 급성 및 만성 단계에 초점을 맞추고 있습니다. 대부분의 행정 연구원이 적용 영구 또는 트란마우스 또는 쥐의 MCA의 지어서 transient 폐색. 환자에서 MCA 폐색의 허혈성 뇌졸중 (4)의 가장 흔한 원인들이다. 필라멘트 모델을 사용하여, MCA 폐색의 근위 외에 원위 MCA 폐색 수술 실험 스트로크 연구 5에서 가장 자주 사용되는 모델은 아마도. MCA 분기합니다 (lenticulo - 선조체 동맥의 분지에) 원위의 폐색은 일반적으로 선조체 여분 주로 신피질에 영향을 미친다. 혈관 폐색은 영구적 또는 일시적 일 수있다. 장기적인 결과에 대해 병소 용적과 매우 낮은 사망률의 재현성이 모델의 주요 이점이다. 여기, 우리는 시상 동에 만성 두개골 창 (CW) 준비 측면을 수행하고, 이후에 어떻게 개두술 방법을 사용하여 창 아래에 말단 행정을 유도 외과 적 방법을 보여줍니다. 이러한 접근법을 통해 허혈성 급성 및 만성 변화 순차 촬상 적용될 수있다, 공 초점 레이저 스캐닝 및 이광자 현미경 생체 내에 에피 - 조명.

타임 라인 대표적인 결과는도 2 및도 3에 도시되어있다. 우수한 시상 동 (도 2 B, C, D) 숙련 된 의사에 의해 수행되는 매우 낮은 사망률과 질병률 레이트 결과 작은 두개 창 폭과 뇌 창 제제. 10 동물 모두 생존, 모든 만성 CW도 이십팔일 수술 후 고품질의 화상을 위해 사용될 수있다. 상처 감염 또는 다른 합병증 문제는 없었다. <p class="j...

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

말단 중간 대뇌 동맥 분기 (MCAO)의 수술 폐색 실험 뇌졸중 연구에서 자주 사용하는 모델입니다. 이 논문은 마우스에서 생체 내에 길이 현미경을위한 기회를 제공하는 측의 두개 창에 삽입, 결합 영구 MCAO의 기본적인 기술을 설명한다.

측면 만성 두개골 창 준비 활성화<em&gt; 생체</em&gt; 마우스의 말초 중간 대뇌 동맥 폐색에 따라 관측

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 ...

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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

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늙은 쥐의 뇌졸중 결과를 연구하기 위해 수정된 경두개 중뇌동맥 폐색 모델

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