저자는 재정 지원에 대 한 아르다 빌 대학 의료 과학 (아르다 빌, 이란)의 연구에 대 한 부 장관에 게 감사 (No를 부여: 9607).

모든 절차는 동물 연구를 실시 위한 아르다 빌 대학교 의료 과학 연구 위원회의 지침에 따라 수행한 (윤리적 ID 번호: 적외선 ARUMS입니다. REC.1394.08).이 시각된 연구에서 사용 하는 성인 남성 Sprague-Dawley 쥐 (300-350 g) 목초지 연구소 (테헤란, 이란)에서 얻은.
1. 마 취와 Flowmetry
<ol><li>4 %isoflurane 사용 하 여 마 취를 유도 하 고 수술 하는 동안 질소 산화물 (70 %v / v)와 산소 (30 %v / v)의 혼합물에 isoflurane (1-1.5%)으로 유지 합니다.</li>
	<li>피드 백 제어 난방 담요에 발생 하기 쉬운 자세에서 anaesthetized 동물을 놓고이 난방 장치에 연결 된 프로브를 직장에 의하여 37±0.5 ° C에서 체온을 유지 합니다.</li>
	<li>건조를 방지 하기 위해 양쪽 눈에 안과 연 고의 작은 금액을 적용 합니다.</li>
	<li>전기 면도기와 두개골의 왼쪽에 외과 영역을 면도. 거 즈 패드와 함께 betadine 솔루션을 사용 하 여 피부를 소독. 린스이 지역 메 마른 패드 70% 에탄올을 포함 하 고 3 주기17두 단계를 반복 합니다.</li>
	<li>진통, 대 수술 지역17에 0.5 %bupivacaine 0.2 mL을 피하 주사.</li>
	<li>(왼쪽된 눈의 측면 법에서의 왼쪽된 귀에 확장 되는) 두개골에 1cm 긴 피부 절 개를 하 고 두개골 노출 왼쪽된 temporalis 근육 해 부. 그런 다음 만들 작은 버 구멍 5mm 측면 하 고 1mm 도플러 유량 계 연필 프로브 팁의 용이성 배치 bregma18 후부.</li>
	<li>경향이 부정사의 위치에서 쥐 바뀌고 레이저 연필 조사 지역 대뇌 혈 류 (rCBF)을 모니터링 하는 데 이전 드릴된 두개골 블라인드 구멍에 넣어.</li>
	<li>각 동물의 초기 rCBF를 기록 하 고 100%로 이것을 고려 하십시오. 특히, 중간 대뇌 동맥 폐색 (MCAO) rCBF의 80%에 감소는 검색된19,20때마다 시작 됩니다.</li>
</ol>2입니다. 초점 Cerebra 허 혈의 유도
<ol><li>목 영역을 면도 하 고 betadine 솔루션 및 70% 에탄올으로 피부를 소독. 진통17수술 사이트에서 피하 0.5 %bupivacaine 0.2 mL를 주사.</li>
	<li>좌 일반 경 동맥 (LCCA) 액세스할 수 목의 복 부 표면에 2 cm 긴 외과 절 개를 확인 합니다. 외부 경 동맥 (ECA) 및 내부 경 동맥 (ICA)의 분기점에 액세스 하려면 인접 근 막과 미주 신경에서 LCCA를 격리 합니다.</li>
	<li>LCCA 또는 5-0 비단 봉합 사를 고용 하는 ECA 영구적으로 선 하 고 ICA pterygopalatine 동맥의 수준에 무료 부.</li>
	<li>느슨하게 LCCA, 주위 5-0 비단 봉합의 넥타이 배치 후 일시적으로 사라지는 혈관 마이크로 클립 클램프.</li>
	<li>이전 배치 느슨한 넥타이 이전 LCCA에 작은 절 개를 하 게 다음 ICA의 luminal 공간에 4-0 실리콘 코팅 나일론 봉합을 삽입 하 고 나일론 봉합 확보 및 혈액 누수 방지 LCCA 주위 봉합 조입니다.</li>
	<li>ICA에서 혈관 마이크로 클립을 제거 합니다. 그런 다음, MCA 근원의 폐색을 나타내는 rCBF에 표시 된 감소를 관찰까지 실리콘 코팅된 intraluminal 필 라 멘 트를 사전. 허 혈 성 기간 (90 분)의 끝에, intraluminal 봉합21철수 여 reperfusion를 시작 합니다.</li>
</ol>3. 경 정 맥 Cannulation 및 에반스 블루 (EB) 주입
<ol><li>1 cm 세로 절 개를 목에는 중간의 왼쪽을 만들고 퉁 명 스럽게 왼쪽된 경 정 맥 (LGV)의 외부 분기 액세스 멀리 표면 근을 해 부. 다음, 영구적으로 5-0 비단 봉합으로 LGV의 두개골 끝 선. 느슨하게 주위 정 맥, 두 관계를 놓고 일시적으로 혈관 마이크로 클램프와 LGV의 심장 끝을 클램프.</li>
	<li>두 봉합 사이 LGV에 작은 절 개를 하 게 그리고 LGV의 luminal 공간으로 가득 heparinized 혈 청 카 테 터 삽입 및 사전 그것은 약 10 m m. 나중, 조여 정 맥 카 테 터를 확보 하 고 출혈 방지 주위 합자. 적은 양의 정 맥 붕괴를 피하기 위해 혈 청을 주입.</li>
	<li>Reperfusion 기간 시작 되는 시점에서 5 분 동안 EB 염료 (식 염 수에 2 %EB 솔루션의 1 mL/kg) 주사 천천히. 그 후,는 정 0.5 mL 정상 식 염 수를 주입 하 여 세척 하 고 주입 정 맥12,22에서 철수.</li>
	<li>마지막으로, 목과 머리의 절 개를 봉합 하 고 intraperitoneally buprenorphine 0.05 mg/kg을 주사 (IP) 수술 후 진통17reperfusion 기간 동안 모든 6-8 h 주사를 반복.</li>
	<li>복구 케이지17수 화를 제공 하기 위해 미리 데워 진된 saline(IP)의 5 mL를 주사.</li>
	<li>장소 특별 한 복구에 동물 온도 및 공기 조화 제어 하 고 마 취에서 동물의 복구를 모니터링 합니다.</li>
</ol>4. 혈액 뇌 장벽 침투성의 평가
<ol><li>Reperfusion의 24 h 후 깊이 thiopental 나트륨과 동물 anesthetize. 그런 다음, 흉 골 아래에 작은 구멍을 만들어 흉 강을 엽니다.</li>
	<li>오른쪽 아 트리 움에 작은 개통을 확인 하 고 미리 따뜻하게 0.9% 식 염 수 (37 ˚C) 안마당에서 정상적인 염 분 출구23무색 될 때까지 순환에서 EB를 씻어 15 분 동안 좌 심 실을 통해 110 mmHg의 압력의 250 mL를 주입.</li>
	<li>즉시 두개골에서 뇌를 제거 하 고 뇌 매트릭스에 배치. 후 각 전구 및 소 뇌를 해 부하 고 중간 선 따라 왼쪽 (Lesioned)에서 뇌의 오른쪽 반구를 분리 후 깨끗 한 면도날을 사용 하 여.</li>
	<li>각 반구의 무게 하 고 별도로 버퍼링 하는 인산 염의 2.5 mL에 균질 trichloroacetic 산 (60%)의 2.5 mL 소용돌이 기계를 사용 하 여 2 분이 혼합.</li>
	<li>1,322 x g에서 30 분 동안 뇌 샘플 원심 고 샘플 10 분 동안 4 ℃ 냉장고에 진정을 허용 합니다.</li>
	<li>610 nm23에 분 광 광도 계에 의해 EB 흡 광도 추정 하는 supernatants를 사용 합니다.</li>
	<li>EB 농도 표준 곡선 및 표현 결과 대 한 뇌 조직의 µ g/g으로 계산 합니다.</li>
</ol>5입니다. 생산 EB 표준 곡선의
<ol><li>EB 버퍼링 하는 인산 염의 5 ml에서의 µ g 1-10의 농도와 EB의 10 샘플 솔루션을 준비 합니다.</li>
	<li>610에 각 샘플의 흡 광도 값 nm는 분 광 광도 계를 사용 하 여.</li>
	<li>Y 축 (그림 1)에 스프레드시트와 X 축에 현재 EB 농도 흡 광도 값을 사용 하 여 분산형 차트를 확인 합니다.</li>
	<li>곡선의 방정식에 대 한 선형 추세선을 정의 합니다. 그런 다음 샘플의 EB 농도를 사용 합니다.</li>
	<li>뇌 조직 무게의 µ g/g으로 EB 넘쳐 흐름의 최종 결과 보고 합니다.</li>
</ol>6. 가짜 작업
<ol><li>가짜 운영 그룹 (포함 절 개 목 지역에 EB 주입)에 쥐와 같은 수술 과정을 수행 하지만 MCAO는 제외.</li>
</ol>

<ol>
	<li>Jin, 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=Protecting+against+cerebrovascular+injury:+contributions+of+12/15-lipoxygenase+to+edema+formation+after+transient+focal+ischemia.">Protecting against cerebrovascular injury: contributions of 12/15-lipoxygenase to edema formation after transient focal ischemia.</a> Stroke. 39 (9), 2538-2543 (2008).</li><li>Lo, E. H., Dalkara, T., Moskowitz, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms,+challenges+and+opportunities+in+stroke.">Mechanisms, challenges and opportunities in stroke.</a> Nat Rev Neurosci. 4 (5), 399-415 (2003).</li><li>Tam, S. J., Watts, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Connecting+vascular+and+nervous+system+development:+angiogenesis+and+the+blood-brain+barrier.">Connecting vascular and nervous system development: angiogenesis and the blood-brain barrier.</a> Annu Rev Neurosci. 33, 379-408 (2010).</li><li>Zhang, C., 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=The+potential+use+of+H102+peptide-loaded+dual-functional+nanoparticles+in+the+treatment+of+Alzheimer's+disease.">The potential use of H102 peptide-loaded dual-functional nanoparticles in the treatment of Alzheimer's disease.</a> J Control Release. , (2014).</li><li>Obermeier, B., Daneman, R., Ransohoff, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development,+maintenance+and+disruption+of+the+blood-brain+barrier.">Development, maintenance and disruption of the blood-brain barrier.</a> Nat Med. 19 (12), 1584-1596 (2013).</li><li>Fang, W., 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=Attenuated+Blood-Brain+Barrier+Dysfunction+by+XQ-1H+Following+Ischemic+Stroke+in+Hyperlipidemic+Rats.">Attenuated Blood-Brain Barrier Dysfunction by XQ-1H Following Ischemic Stroke in Hyperlipidemic Rats.</a> Mol Neurobiol. 52 (1), 162-175 (2015).</li><li>Huang, J., 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=CXCR4+antagonist+AMD3100+protects+blood-brain+barrier+integrity+and+reduces+inflammatory+response+after+focal+ischemia+in+mice.">CXCR4 antagonist AMD3100 protects blood-brain barrier integrity and reduces inflammatory response after focal ischemia in mice.</a> Stroke. 44 (1), 190-197 (2013).</li><li>Omidi, Y., Barar, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impacts+of+blood-brain+barrier+in+drug+delivery+and+targeting+of+brain+tumors.">Impacts of blood-brain barrier in drug delivery and targeting of brain tumors.</a> Bioimpacts. 2 (1), 5-22 (2012).</li><li>Cho, H., 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=Three-dimensional+blood-brain+barrier+model+for+in+vitro+studies+of+neurovascular+pathology.">Three-dimensional blood-brain barrier model for in vitro studies of neurovascular pathology.</a> Sci Rep. 5, (2015).</li><li>Barar, J., Rafi, M. A., Pourseif, M. M., Omidi, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Blood-brain+barrier+transport+machineries+and+targeted+therapy+of+brain+diseases.">Blood-brain barrier transport machineries and targeted therapy of brain diseases.</a> Bioimpacts. 6 (4), 225-248 (2016).</li><li>Kaya, 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=Magnesium+sulfate+attenuates+increased+blood-brain+barrier+permeability+during+insulin-induced+hypoglycemia+in+rats.">Magnesium sulfate attenuates increased blood-brain barrier permeability during insulin-induced hypoglycemia in rats.</a> Can J Physiol Pharmacol. 79 (9), 793-798 (2001).</li><li>Pasban, E., Panahpour, H., Vahdati, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+oxygen+therapy+does+not+protect+the+brain+from+vasogenic+edema+following+acute+ischemic+stroke+in+adult+male+rats.">Early oxygen therapy does not protect the brain from vasogenic edema following acute ischemic stroke in adult male rats.</a> Sci Rep. 7 (1), 3221 (2017).</li><li>Haghnejad Azar, A., Oryan, S., Bohlooli, S., Panahpour, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Alpha-Tocopherol+Reduces+Brain+Edema+and+Protects+Blood-Brain+Barrier+Integrity+following+Focal+Cerebral+Ischemia+in+Rats.">Alpha-Tocopherol Reduces Brain Edema and Protects Blood-Brain Barrier Integrity following Focal Cerebral Ischemia in Rats.</a> Med Princ Pract. 26 (1), 17-22 (2017).</li><li>Belayev, L., Busto, R., Zhao, W., Ginsberg, 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=Quantitative+evaluation+of+blood-brain+barrier+permeability+following+middle+cerebral+artery+occlusion+in+rats.">Quantitative evaluation of blood-brain barrier permeability following middle cerebral artery occlusion in rats.</a> Brain Res. 739 (1-2), 88-96 (1996).</li><li>Bodsch, W., Hossmann, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=125I-antibody+autoradiography+and+peptide+fragments+of+albumin+in+cerebral+edema.">125I-antibody autoradiography and peptide fragments of albumin in cerebral edema.</a> J Neurochem. 41 (1), 239-243 (1983).</li><li>Jiang, Q., 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=Quantitative+evaluation+of+BBB+permeability+after+embolic+stroke+in+rat+using+MRI.">Quantitative evaluation of BBB permeability after embolic stroke in rat using MRI.</a> J Cereb Blood FlowMetab. 25 (5), 583-592 (2005).</li><li>Ulu&#231;, K., Miranpuri, A., Kujoth, G. C., Akt&#252;re, E., Ba&#351;kaya, M. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Focal+cerebral+ischemia+model+by+endovascular+suture+occlusion+of+the+middle+cerebral+artery+in+the+rat.">Focal cerebral ischemia model by endovascular suture occlusion of the middle cerebral artery in the rat.</a> J Vis Exp. (48), (2011).</li><li>Hungerhuber, E., Zausinger, S., Westermaier, T., Plesnila, N., Schmid-Elsaesser, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simultaneous+bilateral+laser+Doppler+fluxmetry+and+electrophysiological+recording+during+middle+cerebral+artery+occlusion+in+rats.">Simultaneous bilateral laser Doppler fluxmetry and electrophysiological recording during middle cerebral artery occlusion in rats.</a> J Neurosci Methods. 154 (1-2), 109-115 (2006).</li><li>Panahpour, H., Nouri, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Post-Ischemic+Treatment+with+candesartan+protect+from+cerebral+ischemic/reperfusioninjury+in+normotensive+rats.">Post-Ischemic Treatment with candesartan protect from cerebral ischemic/reperfusioninjury in normotensive rats.</a> Int J Pharm Pharm Sci. 4 (4), 286-289 (2012).</li><li>Panahpour, H., Dehghani, G. A., Bohlooli, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enalapril+attenuates+ischaemic+brain+oedema+and+protects+the+blood-brain+barrier+in+rats+via+an+anti-oxidant+action.">Enalapril attenuates ischaemic brain oedema and protects the blood-brain barrier in rats via an anti-oxidant action.</a> Clin Exp Pharmacol Physiol. 41 (3), 220-226 (2014).</li><li>Panahpour, H., Nekooeian, A. A., Dehghani, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Blockade+of+Central+Angiotensin+II+AT1+Receptor+Protects+the+Brain+from+Ischemia/Reperfusion+Injury+in+Normotensive+Rats.">Blockade of Central Angiotensin II AT1 Receptor Protects the Brain from Ischemia/Reperfusion Injury in Normotensive Rats.</a> Iran J Med Sci. 39 (6), 536-542 (2014).</li><li>Panahpour, H., Nekooeian, A. A., Dehghani, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Candesartan+attenuates+ischemic+brain+edema+and+protects+the+blood-brain+barrier+integrity+from+ischemia/reperfusion+injury+in+rats.">Candesartan attenuates ischemic brain edema and protects the blood-brain barrier integrity from ischemia/reperfusion injury in rats.</a> Iran Biomed J. 18 (4), 232-238 (2014).</li><li>Kaya, 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=The+effects+of+magnesium+sulfate+on+blood-brain+barrier+disruption+caused+by+intracarotid+injection+of+hyperosmolar+mannitol+in+rats.">The effects of magnesium sulfate on blood-brain barrier disruption caused by intracarotid injection of hyperosmolar mannitol in rats.</a> Life sci. 76 (2), 201-212 (2004).</li><li>Sch&#246;ller, K., 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=Characterization+of+microvascular+basal+lamina+damage+and+blood-brain+barrier+dysfunction+following+subarachnoid+hemorrhage+in+rats.">Characterization of microvascular basal lamina damage and blood-brain barrier dysfunction following subarachnoid hemorrhage in rats.</a> Brain Res. 1142, 237-246 (2007).</li><li>Bodsch, W., Hossmann, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=125I-Antibody+Autoradiography+and+Peptide+Fragments+of+Albumin+in+Cerebral+Edema.">125I-Antibody Autoradiography and Peptide Fragments of Albumin in Cerebral Edema.</a> J Neurochem. 41 (1), 239-243 (1983).</li><li>Sandoval, K. E., Witt, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Blood-brain+barrier+tight+junction+permeability+and+ischemic+stroke.">Blood-brain barrier tight junction permeability and ischemic stroke.</a> Neurobiol Dis. 32 (2), 200-219 (2008).</li><li>Zhu, H., 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=Baicalin+reduces+the+permeability+of+the+blood-brain+barrier+during+hypoxia+in+vitro+by+increasing+the+expression+of+tight+junction+proteins+in+brain+microvascular+endothelial+cells.">Baicalin reduces the permeability of the blood-brain barrier during hypoxia in vitro by increasing the expression of tight junction proteins in brain microvascular endothelial cells.</a> J Ethnopharmacol. 141 (2), 714-720 (2012).</li><li>Kucuk, 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=Effects+of+losartan+on+the+blood-brain+barrier+permeability+in+long-term+nitric+oxide+blockade-induced+hypertensive+rats.">Effects of losartan on the blood-brain barrier permeability in long-term nitric oxide blockade-induced hypertensive rats.</a> Life Sci. 71 (8), 937-946 (2002).</li><li>Uyama, O., 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=Quantitative+evaluation+of+vascular+permeability+in+the+gerbil+brain+after+transient+ischemia+using+Evans+blue+fluorescence.">Quantitative evaluation of vascular permeability in the gerbil brain after transient ischemia using Evans blue fluorescence.</a> J Cereb Blood Flow Metab. 8 (2), 282-284 (1988).</li><li>Kleinig, T. J., Vink, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Suppression+of+inflammation+in+ischemic+and+hemorrhagic+stroke:+therapeutic+options.">Suppression of inflammation in ischemic and hemorrhagic stroke: therapeutic options.</a> Curr Opin Neurol. 22 (3), 294-301 (2009).</li><li>Del Zoppo, G. J., Mabuchi, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cerebral+microvessel+responses+to+focal+ischemia.">Cerebral microvessel responses to focal ischemia.</a> J Cereb Blood Flow Metab. 23 (8), 879-894 (2003).</li><li>Fluri, F., Schuhmann, M. K., Kleinschnitz, C. <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+ischemic+stroke+and+their+application+in+clinical+research.">Animal models of ischemic stroke and their application in clinical research.</a> Drug Des Devel Ther. 9, 3445-3454 (2015).</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>Shin, H. K., 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=Mild+induced+hypertension+improves+blood+flow+and+oxygen+metabolism+in+transient+focal+cerebral+ischemia.">Mild induced hypertension improves blood flow and oxygen metabolism in transient focal cerebral ischemia.</a> Stroke. 39 (5), 1548-1555 (2008).</li><li>Bottiger, B. W., 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=Global+cerebral+ischemia+due+to+cardiocirculatory+arrest+in+mice+causes+neuronal+degeneration+and+early+induction+of+transcription+factor+genes+in+the+hippocampus.">Global cerebral ischemia due to cardiocirculatory arrest in mice causes neuronal degeneration and early induction of transcription factor genes in the hippocampus.</a> Brain Res Mol Brain Res. 65 (2), 135-142 (1999).</li></ol>

저자는 공개 없다.

지금까지, autoradiography 방사성 추적기24,25, 면역 형광 검사 현미경 검사 법26,27, EB 넘쳐 흐름 기술은20, 의 검출 등 다양 한 방법 23 혈액-뇌 장벽 손상을 평가 하는 데 사용 되었습니다. EB 염료 강력 하 게 혈 청 알 부 민에 바인딩할 수 이며 혈관 누설을 감지 하 고 측?...

Vasogenic 뇌 부 종 혈액-뇌 장벽 (BBB) 중단으로 인해 허 혈 성 뇌졸중의 중요 한 합병증 및 뇌졸중 환자1,2에서 생존 율의 주요 결정 요인이 남아 있다. 혈액-뇌 장벽 (BBB)는 뇌 모 세관 내 피 세포 (BCECs)에 의해 형성 된 뚜렷한 혈관 구성 요소 (예: BCECs, pericytes, astroglial, 및 신경 세포3중 꽉 접합)의 구성, 제공 하는 중앙 신경 조직 (CNS) 말 초 혈액 순환4,5사이의 특수 하 고 역동적인 인터페이스. 국 소 빈 혈 reperfusion 상해 등 모욕 BBB의 기능적 무결성을 방해 하 고 궁극적으로 트리거 대뇌 염증 및 기본 뇌 손상 뇌 실질에 순환 하는 백혈구의 침투를 후속으로 이어질 수 있습니다. 6 , 7. 뇌졸중의 발생에 따라 BBB의 부전의 정확한 검출을 위한 동물 모델 필요. 이러한 모델은 새로운 신경 보호 전략을 소개 하 고 기본 병 태 생리 기계 장치 공부에 대 한 매우 중요. 체 외에서 세포 문화 기반 모델 BBB의 높은 개발 하 고 되었습니다 BBB physiopathology8,,910의 분자 연구에 사용. 그럼에도 불구 하 고, vivo에서 동물 모델, 인간의 임상 조건에 비슷한 BBB의 허 혈 성 손상, 생산은 또한 매우 보람 이와. 에반스 블루 (EB)의 넘쳐 흐름의 정량적 검출은 잘 허용 하 고 민감한 기술 평가 BBB 무결성 및 신경 퇴행 성 질환, 허 혈 성 뇌졸중11, 를 포함 하 여 기능에 대 한 사용 되었습니다. 12 , 13 , 14.이 방법은 비용, 재현성, 및 완전히 적용 가능한 모든 실험 실험실에서. 그것의 구현 방사성 트레이 서15 또는 자기 공명 영상 (MRI)16다른 방법에 대 한 필수 구성 요소를 같은 고급 장비를 필요 하지 않습니다. 이 문서에서는, BBB 평가 EB 넘쳐 흐름을 사용 하 여 허 혈 성 뇌졸중의 쥐 모델에서의 기본적인 기술적 프로세스 포괄적으로 설명 합니다.

<table border="0" cellpadding="0" cellspacing="0" width="829">
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			<td>Sigma-Aldrich&#160;</td>
			<td>64-17-5</td>
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			<td height="22" style="height:22px;">silicone(Xantopren)</td>
			<td>Heraeus</td>
			<td>EN ISO 4823</td>
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			<td height="22" style="height:22px;">Activator universal plus</td>
			<td>Heraeus</td>
			<td>66037445</td>
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			<td height="22" style="height:22px;">Micro-Dissecting forceps</td>
			<td>Stoelting</td>
			<td>52100-41</td>
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			<td>Stoelting</td>
			<td>52130-00</td>
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			<td>&#160;Kent Scientific</td>
			<td>SS-01</td>
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			<td height="22" style="height:22px;">Power Lab system</td>
			<td>AD Instruments</td>
			<td>ML880</td>
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			<td height="22" style="height:22px;">Laser Doppler flowmeter</td>
			<td>AD Instruments</td>
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			<td>Harvard Appratus</td>
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			<td>FineScience Tools</td>
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			<td height="22" style="height:22px;">Silk 5-0 suture thread</td>
			<td>Ethicon</td>
			<td>682G</td>
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			<td height="22" style="height:22px;">Ethilon 4-0 suture thread&#160;</td>
			<td>Ethicon</td>
			<td>EH6740G</td>
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an in vivo assessment of blood-brain barrier disruption in a rat model of ischemic stroke

허 혈 성 뇌졸중 vasogenic 대뇌 부 종 및 후속 기본 뇌 손상, 혈액-뇌 장벽 (BBB)의 파괴를 통해 중재 된다 이끌어 낸다. 유발 허 혈 성 뇌졸중 쥐 설립 되었고 BBB의 기능 무결성을 조사 하 비보에 모델로 사용. 허 혈 성 상해 두뇌 샘플에서 에반스 블루 (EB)의 spectrophotometric 감지 소설 치료 modalities의 개발과 연구에 대 한 신뢰할 수 있는 정당성을 제공할 수 있습니다. 이 방법은 재현성 결과 생성 하 고 특별 한 장비를 위한 필요 없이 어떤 실험실에 적용 됩니다. 여기, 선물이 쥐에서 허 혈 성 뇌졸중의 유도 따라 EB의 넘쳐 흐름의 검출에 시각 및 기술 지침.

가짜 운영 쥐의 좌 반구와 우 반구에서 EB 수준에 중요 한 차이가 없었다 (1.06 ± 0.1 µ g/g과 1.1 ± 0.09 µ g/g, 각각). 그림 2A 2B, 일시적인 허 혈의 유도에서 같이 (90 분 허 혈 / 24 h reperfusion) EB 수준에 상당한 차이 발생 (10.41 ± 0.84 µ g/g, p < 0.001) 각각에 비해 허 혈 성 쥐의 왼쪽된 반구에 운영 하는 가짜 쥐에 반구입니다. 샨 다, 이러한 결과 나타냅니다 정상적?...

Watch this Scientific Journal Video about An In Vivo Assessment of Blood-Brain Barrier Disruption in a Rat Model of Ischemic Stroke at JoVE.com

An In Vivo Assessment of Blood-Brain Barrier Disruption in a Rat Model of Ischemic Stroke

허 혈 성 뇌졸중의 쥐 모델에서 뇌혈관 장애의 비보에 평가

이 절차의 전반적인 목표는 비보에 허 혈 성 뇌졸중의 쥐 모델에서 뇌혈관 장애의 평가 대 한 높은 재현 기법을 제공 하는입니다.

Ischemic stroke leads to vasogenic cerebral edema and subsequent primary brain injury, which is mediated through destruction of the blood-brain barrier ...

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Neuroscience

An In Vivo Assessment of Blood-Brain Barrier Disruption in a Rat Model of Ischemic Stroke

11.3K 조회수.  Ardabil University of Medical Sciences. 이 절차의 전반적인 목표는 비보에 허 혈 성 뇌졸중의 쥐 모델에서 뇌혈관 장애의 평가 대 한 높은 재현 기법을 제공 하는입니다.

비디오: An In Vivo Assessment of Blood-Brain Barrier Disruption in a Rat Model of Ischemic Stroke

Generation of an Immortalized Murine Brain Microvascular Endothelial Cell Line as an In Vitro Blood Brain Barrier Model

Epithelial and endothelial cells (EC) are building paracellular barriers which protect the tissue from the external and internal environment. The blood-brain barrier (BBB) consisting of EC, astrocyte end-feet, pericytes and the basal membrane is responsible for the protection and homeostasis of the brain parenchyma. In vitro BBB models are common tools to study the structure and function of the BBB at the cellular level. A considerable number of different in vitro BBB models have been established for research in different laboratories to date. Usually, the cells are obtained from bovine, porcine, rat or mouse brain tissue (discussed in detail in the review by Wilhelm et al. 1). Human tissue samples are available only in a restricted number of laboratories or companies 2,3. While primary cell preparations are time consuming and the EC cultures can differ from batch to batch, the establishment of immortalized EC lines is the focus of scientific interest.
Here, we present a method for establishing an immortalized brain microvascular EC line from neonatal mouse brain. We describe the procedure step-by-step listing the reagents and solutions used. The method established by our lab allows the isolation of a homogenous immortalized endothelial cell line within four to five weeks. The brain microvascular endothelial cell lines termed cEND 4 (from cerebral cortex) and cerebEND 5 (from cerebellar cortex), were isolated according to this procedure in the F&ouml;rster laboratory and have been effectively used for explanation of different physiological and pathological processes at the BBB. Using cEND and cerebEND we have demonstrated that these cells respond to glucocorticoid- 4,6-9 and estrogen-treatment 10 as well as to pro-infammatory mediators, such as TNFalpha 5,8. Moreover, we have studied the pathology of multiple sclerosis 11 and hypoxia 12,13 on the EC-level. The cEND and cerebEND lines can be considered as a good tool for studying the structure and function of the BBB, cellular responses of ECs to different stimuli or interaction of the EC with lymphocytes or cancer cells.

Generation of an Immortalized Murine Brain Microvascular Endothelial Cell Line as an In Vitro Blood Brain Barrier Model

This method describes how to isolate and immortalize microvascular endothelial cells from mouse brain. We describe a step-by-step protocol starting from the homogenization of brain tissue, digestion steps, seeding and immortalization of the cells. Usually, it takes about five weeks to obtain a homogenous, immortalized microvascular endothelial cell line.

로 불후의 손쥐 뇌 Microvascular 내피 세포 라인의 생성<em&gt; 체외에서</em&gt; 혈액 뇌 장벽 모델

Epithelial and endothelial cells (EC) are building paracellular barriers which protect the tissue from the external and internal environment. The ...

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신경과학

Functional Neuroimaging Using Ultrasonic Blood-brain Barrier Disruption and Manganese-enhanced MRI

Although mice are the dominant model system for studying the genetic and molecular underpinnings of neuroscience, functional neuroimaging in mice remains technically challenging. One approach, Activation-Induced Manganese-enhanced MRI (AIM MRI), has been used successfully to map neuronal activity in rodents 1-5. In AIM MRI, Mn2+ acts a calcium analog and accumulates in depolarized neurons 6,7. Because Mn2+ shortens the T1 tissue property, regions of elevated neuronal activity will enhance in MRI. Furthermore, Mn2+ clears slowly from the activated regions; therefore, stimulation can be performed outside the magnet prior to imaging, enabling greater experimental flexibility. However, because Mn2+ does not readily cross the blood-brain barrier (BBB), the need to open the BBB has limited the use of AIM MRI, especially in mice.
One tool for opening the BBB is ultrasound. Though potentially damaging, if ultrasound is administered in combination with gas-filled microbubbles (i.e., ultrasound contrast agents), the acoustic pressure required for BBB opening is considerably lower. This combination of ultrasound and microbubbles can be used to reliably open the BBB without causing tissue damage 8-11.
Here, a method is presented for performing AIM MRI by using microbubbles and ultrasound to open the BBB. After an intravenous injection of perflutren microbubbles, an unfocused pulsed ultrasound beam is applied to the shaved mouse head for 3 minutes. For simplicity, we refer to this technique of BBB Opening with Microbubbles and UltraSound as BOMUS 12. Using BOMUS to open the BBB throughout both cerebral hemispheres, manganese is administered to the whole mouse brain. After experimental stimulation of the lightly sedated mice, AIM MRI is used to map the neuronal response.
To demonstrate this approach, herein BOMUS and AIM MRI are used to map unilateral mechanical stimulation of the vibrissae in lightly sedated mice 13. Because BOMUS can open the BBB throughout both hemispheres, the unstimulated side of the brain is used to control for nonspecific background stimulation. The resultant 3D activation map agrees well with published representations of the vibrissae regions of the barrel field cortex 14. The ultrasonic opening of the BBB is fast, noninvasive, and reversible; and thus this approach is suitable for high-throughput and/or longitudinal studies in awake mice.

A technique is described for broadly opening the blood-brain barrier in the mouse using microbubbles and ultrasound. Using this technique, manganese can be administered to the mouse brain. Because manganese is an MRI contrast agent that accumulates in depolarized neurons, this approach enables imaging of neuronal activity.

초음파 혈액 - 뇌 장벽 파괴 및 망간 향상된 MRI를 사용하여 기능성 Neuroimaging

Although mice are the dominant model system for studying the genetic and molecular underpinnings of neuroscience, functional neuroimaging in mice remains ...

Assessment of Blood-brain Barrier Permeability by Intravenous Infusion of FITC-labeled Albumin in a Mouse Model of Neurodegenerative Disease

Disruption of blood-brain barrier (BBB) integrity is a common feature for different neurological and neurodegenerative diseases. Although the interplay between perturbed BBB homeostasis and the pathogenesis of brain disorders needs further investigation, the development and validation of a reliable procedure to accurately detect BBB alterations may be crucial and represent a useful tool for potentially predicting disease progression and developing targeted therapeutic strategies.
Here, we present an easy and efficient procedure for evaluating BBB leakage in a neurodegenerative condition like that occurring in a preclinical mouse model of Huntington disease, in which defects in the permeability of BBB are clearly detectable precociously in the disease. Specifically, the high molecular weight fluorescein isothiocyanate labelled (FITC)-albumin, which is able to cross the BBB only when the latter is impaired, is acutely infused into a mouse jugular vein and its distribution in the vascular or parenchymal districts is then determined by fluorescence microscopy.
Accumulation of green fluorescent-albumin in the brain parenchyma functions as an index of aberrant BBB permeability and, when quantitated by using Image J processing software, is reported as Green Fluorescence Intensity.

In this study, we present an easy and efficient procedure for evaluating the disruption of the blood-brain barrier under neurodegenerative conditions. To achieve our objective, we infused high molecular weight fluorescein isothiocyanate labelled (FITC)-albumin into mouse jugular vein and evaluated its leakage into the brain parenchyma by fluorescence microscopy.

FITC 표시 된 알 부 민 신경 질환의 마우스 모델에서의 정 맥 주입에 의해 혈액-뇌 장벽 침투성의 평가

Disruption of blood-brain barrier (BBB) integrity is a common feature for different neurological and neurodegenerative diseases. Although the interplay ...

Optimized Management of Endovascular Treatment for Acute Ischemic Stroke

This manuscript describes a streamlined protocol for the management of patients with acute ischemic stroke, which aims at the minimization of time from hospital admission to reperfusion. Rapid restoration of cerebral blood flow is essential for the outcomes of patients with acute ischemic stroke. Endovascular treatment (EVT) has become the standard of care to accomplish this in patients with acute stroke due to large vessel occlusion (LVO). To achieve reperfusion of ischemic brain regions as fast as possible, all in-hospital time delays have to be carefully avoided. Therefore, management of patients with acute ischemic stroke was optimized with an interdisciplinary standard operating procedure (SOP). Stroke neurologists, diagnostic as well as interventional neuroradiologists, and anesthesiologists streamlined all necessary processes from patient admission and diagnosis to EVT of eligible patients. Target times for every step were established. Actually achieved times were prospectively recorded along with clinical data and imaging scores for all endovascularly treated stroke patients. These data were regularly analyzed and discussed in interdisciplinary team meetings. Potential issues were evaluated and all staff involved was trained to adhere to the SOP. This streamlined patient management approach and enhanced interdisciplinary collaboration reduced time from patient admission to reperfusion significantly and was accompanied by a beneficial effect on clinical outcomes.

The outcome of patients with acute ischemic stroke depends on swift restoration of cerebral blood flow. This protocol aims at optimizing the management of such patients by minimizing peri-procedural timings and rendering the time from hospital admission to reperfusion as short as possible.

급성 허 혈 성 뇌졸중 혈관 내 수술 치료의 최적화 관리

This manuscript describes a streamlined protocol for the management of patients with acute ischemic stroke, which aims at the minimization of time from ...

An In Vivo Blood-brain Barrier Permeability Assay in Mice Using Fluorescently Labeled Tracers

Blood-brain barrier (BBB) is a specialized barrier that protects the brain microenvironment from toxins and pathogens in the circulation and maintains brain homeostasis. The principal sites of the barrier are endothelial cells of the brain capillaries whose barrier function results from tight intercellular junctions and efflux transporters expressed on the plasma membrane. This function is regulated by pericytes and astrocytes that together form the neurovascular unit (NVU). Several neurological diseases such as stroke, Alzheimer&#39;s disease (AD), brain tumors are associated with an impaired BBB function. Assessment of the BBB permeability is therefore crucial in evaluating the severity of the neurological disease and the success of the treatment strategies employed.
We present here a simple yet robust permeability assay that have been successfully applied to several mouse models both, genetic and experimental. The method is highly quantitative and objective in comparison to the tracer fluorescence analysis by microscopy that is commonly applied. In this method, mice are injected intraperitoneally with a mix of aqueous inert fluorescent tracers followed by anesthetizing the mice. Cardiac perfusion of the animals is performed prior to harvesting brain, kidneys or other organs. Organs are homogenized and centrifuged followed by fluorescence measurement from the supernatant. Blood drawn from the cardiac puncture just before perfusion serves for normalization purpose to the vascular compartment. The tissue fluorescence is normalized to the wet weight and serum fluorescence to obtain a quantitative tracer permeability index. For additional confirmation, the contralateral hemi-brain preserved for immunohistochemistry can be utilized for tracer fluorescence visualization purposes.

An In Vivo Blood-brain Barrier Permeability Assay in Mice Using Fluorescently Labeled Tracers

Here we present a mouse brain vascular permeability assay using intraperitoneal injection of fluorescent tracers followed by perfusion that is applicable to animal models of blood-brain barrier dysfunction. One hemi-brain is used for assessing permeability quantitatively and the other for tracer visualization/immunostaining. The procedure takes 5 - 6 h for 10 mice.

Vivo에서 혈액-뇌 장벽 침투성 분석 결과 사용 하 여 마우스에 붙일 추적기 표시

Blood-brain barrier (BBB) is a specialized barrier that protects the brain microenvironment from toxins and pathogens in the circulation and maintains ...

The Power of Interstimulus Interval for the Assessment of Temporal Processing in Rodents

Temporal processing deficits have been implicated as a potential elemental dimension of higher-level cognitive processes, commonly observed in neurocognitive disorders. Despite the popularization of prepulse inhibition (PPI) in recent years, many current protocols promote using a percent of control measure, thereby precluding the assessment of temporal processing. The present study used cross-modal PPI and gap prepulse inhibition (gap-PPI) to demonstrate the benefits of employing a range of interstimulus intervals (ISIs) to delineate effects of sensory modality, psychostimulant exposure, and age. Assessment of sensory modality, psychostimulant exposure, and age reveals the utility of an approach varying the interstimulus interval (ISI) to establish the shape of the ISI function, including increases (sharper curve inflections) or decreases (flattening of the response amplitude curve) in startle amplitude. Additionally, shifts in peak response inhibition, suggestive of a differential sensitivity to the manipulation of ISI, are often revealed. Thus, the systematic manipulation of ISI affords a critical opportunity to evaluate temporal processing, which may reveal the underlying neural mechanisms involved in neurocognitive disorders.

Temporal processing, a preattentive process, may underlie deficits in higher-level cognitive processes, including attention, commonly observed in neurocognitive disorders. Using prepulse inhibition as an exemplar paradigm, we present a protocol for manipulating interstimulus interval (ISI) to establish the shape of the ISI function to provide an assessment of temporal processing.

설치류에서 시간적 처리의 평가 대 한 Interstimulus 간격의 힘

Temporal processing deficits have been implicated as a potential elemental dimension of higher-level cognitive processes, commonly observed in ...

A Preclinical Model to Assess Brain Recovery After Acute Stroke in Rats

The purpose of this study was to establish and validate an animal brain ischemia model in the recovery and sequela stages. A middle cerebral artery occlusion/reperfusion (MCAO/R) model in male Sprague-Dawley rats was chosen. By changing the rat&#39;s weight (260&#8722;330 g), the thread bolt type (2636/2838/3040/3043) and the brain infarct time (2-3 h), a higher Longa&#39;s score, a larger infarct volume and a greater model success ratio were screened using the Longa&#39;s score and TTC staining. The optimum model condition (300 g, 3040 thread bolt, 3 h brain infarct time) was acquired and used in a 1-90 day observation period after reperfusion via assessment of sensorimotor functions and infarct volume. At these conditions, the bilateral asymmetry test had a significant difference from 1 to 90 days, and the grid-walking test had a significant difference from 1 to 60 days; both differences could be a suitable sensorimotor functional test. Thus, the most appropriate condition of a novel rat model in the recovery and sequela stages of brain ischemia was found: 300 g rats that underwent MCAO with a 3040 thread bolt for a 3 h brain infarct and then reperfused. The appropriate sensorimotor functional tests were a bilateral asymmetry test and a grid-walking test.

The&#160;purpose of this study is to establish and validate an animal model for research in the recovery and sequela stages of brain ischemia by testing brain infarction and sensorimotor function after middle cerebral artery occlusion/reperfusion (MCAO/R) after 1-90 days in rats.

쥐에서 급성 뇌졸중 후 뇌 회복을 평가하는 전임상 모델

The purpose of this study was to establish and validate an animal brain ischemia model in the recovery and sequela stages. A middle cerebral artery ...

A Benchtop Approach to the Location Specific Blood Brain Barrier Opening using Focused Ultrasound in a Rat Model

Stereotaxic surgery is the gold standard for localized drug and gene delivery to the rodent brain. This technique has many advantages over systemic delivery including precise localization to a target brain region and reduction of off target side effects. However, stereotaxic surgery is highly invasive which limits its translational efficacy, requires long recovery times, and provides challenges when targeting multiple brain regions. Focused ultrasound (FUS) can be used in combination with circulating microbubbles to transiently open the blood brain barrier (BBB) in millimeter sized regions. This allows intracranial localization of systemically delivered agents that cannot normally cross the BBB. This technique provides a noninvasive alternative to stereotaxic surgery. However, to date this technique has yet to be widely adopted in neuroscience laboratories due to the limited access to equipment and standardized methods. The overall goal of this protocol is to provide a benchtop approach to FUS BBB opening (BBBO) that is affordable and reproducible and can therefore be easily adopted by any laboratory.

Focused ultrasound with microbubble agents can open the blood brain barrier focally and transiently. This technique has been used to deliver a wide range of agents across the blood brain barrier. This article provides a detailed protocol for the localized delivery to the rodent brain with or without MRI guidance.

쥐 모델에서 집중 초음파를 사용하여 위치 특정 혈액 뇌 장벽 개방에 대한 벤치 탑 접근

Stereotaxic surgery is the gold standard for localized drug and gene delivery to the rodent brain. This technique has many advantages over systemic ...

Measuring Post-Stroke Cerebral Edema, Infarct Zone and Blood-Brain Barrier Breakdown in a Single Set of Rodent Brain Samples

One of the most common causes of morbidity and mortality worldwide is ischemic stroke. Historically, an animal model used to stimulate ischemic stroke involves middle cerebral artery occlusion (MCAO). Infarct zone, brain edema and blood-brain barrier (BBB) breakdown are measured as parameters that reflect the extent of brain injury after MCAO. A significant limitation to this method is that these measurements are normally obtained in different rat brain samples, leading to ethical and financial burdens due to the large number of rats that need to be euthanized for an appropriate sample size. Here we present a method to accurately assess brain injury following MCAO by measuring infarct zone, brain edema and BBB permeability in the same set of rat brains. This novel technique provides a more efficient way to evaluate the pathophysiology of stroke.

This protocol describes a novel technique of measuring the three most important parameters of ischemic brain injury on the same set of rodent brain samples. Using only one brain sample is highly advantageous in terms of ethical and economic costs.

설치류 뇌 샘플의 단일 세트에서 뇌졸중 후 뇌 부종, 경색 영역 및 혈액 뇌 장벽 고장 측정

One of the most common causes of morbidity and mortality worldwide is ischemic stroke. Historically, an animal model used to stimulate ischemic stroke ...

A Rat Model of EcoHIV Brain Infection

It has been well studied that the EcoHIV infected mouse model is of significant utility in investigating HIV associated neurological complications. Establishment of the EcoHIV infected rat model for studies of drug abuse and neurocognitive disorders, would be beneficial in the study of neuroHIV and HIV-1 associated neurocognitive disorders (HAND). In the present study, we demonstrate the successful creation of a rat model of active HIV infection using chimeric HIV (EcoHIV). First, the lentiviral construct of EcoHIV was packaged in cultured 293 FT cells for 48 hours. Then, the conditional medium was concentrated and titered. Next, we performed bilateral stereotaxic injections of the EcoHIV-EGFP into F344/N rat brain tissue. One week after infection, EGFP fluorescence signals were detected in the infected brain tissue, indicating that EcoHIV successfully induces an active HIV infection in rats. In addition, immunostaining for the microglial cell marker, Iba1, was performed. The results indicated that microglia were the predominant cell type harboring EcoHIV. Furthermore, EcoHIV rats exhibited alterations in temporal processing, a potential underlying neurobehavioral mechanism of HAND as well as synaptic dysfunction eight weeks after infection. Collectively, the present study extends the EcoHIV model of HIV-1 infection to the rat offering a valuable biological system to study HIV-1 viral reservoirs in the brain as well as HAND and associated comorbidities such as drug abuse.

Here, we present a protocol to establish a new rat model of active HIV infection using chimeric HIV (EcoHIV), which is critical for enhancing our understanding of HIV-1 viral reservoirs in the brain and offering a system to study HIV-associated neurocognitive disorders and associated comorbidities (i.e., drug abuse).

에코HIV 뇌 감염의 쥐 모델

It has been well studied that the EcoHIV infected mouse model is of significant utility in investigating HIV associated neurological complications. ...