Name: a repetitive concussive head injury model in mice
Uploaded: 2016-10-12T14:00:00.000+00:00
Duration: 5 min 42 s
Description: Watch this Scientific Journal Video about A Repetitive Concussive Head Injury Model in Mice at JoVE.com

This works was supported by funding from a Florida Health grant (Brain and spinal cord injury research fund) (KKW).

모든 절차는 플로리다 대학의 기관 동물 관리 및 사용위원회 및 실험 동물의 관리 및 사용을위한 건강 가이드의 국립 연구소에 따라 승인 된 프로토콜 번호 201207692에서 수행 하였다. 1. 동물 관리 <ol><li> 3-4 개월 된 수컷 C57BL / 6J 마우스를 사용합니다. 침구, 중첩 재료, 음식과 임의 량의 물을 제공한다. 일정 12 시간 빛 / 12 시간 다크 사이클 22 ° C - 20 제어 주위 온도에 마우스를 유지합니다. </li></ol> 2. 사전 매복 준비 <ol><li> 전자 정위 충격 장치에 맞춤형 실리콘 고무 코팅 금속 끝을 연결합니다. 팁의 평평한 바닥이 프로브 팁 (그림 1A)의 표면에 평행해야합니다. </li><li> 2.5 %의 이소 플루 란의 유지 보수 마취 다음에 4 % 이소 플루 란과 마우스를 마취. 유량계를 통해 마취를 확인합니다. 과정 제어R 동물까지 마취 수준은 페달 철수 반사의 손실을 보여줌으로써 마취의 수술 수준에 도달. </li><li> 가열 패드에 발생하기 쉬운 위치에 마우스를 놓습니다. 마취 마우스를 유지하기 위해 깔때기 모양의 코 콘을 사용합니다. 완전히 트리머를 사용하여 머리를 면도. 마취 동안 건조를 방지하기 위해 마우스의 눈에 바셀린 안과 연고를 사용합니다. </li></ol> 3. 충격 매개 변수 설정 주 : 충격 시스템은 충격 파라미터를 설정하는 제어 박스의 박힘을 수행하기위한 액츄에이터, 및 3 운동 축을 디지털 정위 프레임을 포함한다. <ol><li> 4m / 초 충격 장치의 속도를 미리 설정하고 제어 박스에 240 msec의 시간 드웰. </li></ol> 4. 위치 충격 센터 <ol><li> 39 ° C 부근의 체온을 유지하기 위해 동물의 신체에 따라 소프트 가열 패드를 넣는다. 포르노에 정위 프레임에 마우스를 장착무딘 엔드 귀 막대 전자 위치입니다. </li><li> Z 축 드라이버를 이동하여 마우스의 머리에 가까운 충격 팁을 낮 춥니 다. 대상 시상 봉합 위 좌표의 중간 X- 및 Y 드라이버를 이동시켜 평면 영향 팁 (9 mm 직경)을 조정한다. </li><li> 상하 두 귀 (도 1C) 사이에 그려진 가상 수평선에 평행 한 충돌 팁 확실히 한쪽 가장자리를 만든다. 충격의 중심 (0 mm, 측면 4.5 mm를 간가에 간가 9mm) interfrontal과 lambdoid 봉합 사이의 중앙 시상 봉합의 중간에 해당한다. </li></ol> 5. 충격 깊이 설정 <ol><li> 정확하게 타격 깊이를 설정하기 위해, 절연 된 실리콘 고무 코팅 충격 팁을 교체 추가 탐침을 사용한다. </li><li> 팁을 전환하기 전에 제로로 디지털 정위 제어판의 X와 Y의 채널을 설정 전환 팁 후 충격의 중심에는 변화가 없다 확인하십시오. </li><li> PROB 이동수동 X 및 Y 축 모터를 이동하여 충격 영역의 중심에 예를 팁. </li><li> 마우스의 꼬리에 클립 접촉 센서. </li><li> 프로브 팁의 충격 부위의 표면에 닿을 때까지 임팩터 (Z 드라이브)를 이동. </li><li> 제로로 정위 제어판의 Z 채널을 설정합니다. </li><li> 수동 X- 및 Y 드라이버까지 X 및 Y 드라이버 (NOT 디지털 정위 조작부 제로 버튼)을 조정하여 다시 충돌 면적에 영향 팁을 이동 (충격 팁 나란히 배치 된)는 0이다. </li><li> 제어 박스에 인입 스위치를 이동시킴으로써, 액추에이터 후퇴. 수동 4mm에 의해 아래로 임팩터 (Z 드라이버)를 이동합니다. </li></ol> 6. 영향 <ol><li> 컨트롤 박스에 대한 영향 스위치를 눌러 영향을 트리거 4mm의 변형 깊이를 달성한다. </li></ol> 7. 후 매복 <ol><li> 마우스의 첫 호흡 타이머를 사용하여 충격까지의 시간을 측정한다. </li><li&gt; 악기에서 동물을 제거하고 체온을 유지하기 위해 따뜻한 패드에 배치. 이 흉골 드러 누움을 유지하기 위해 충분한 의식을 회복 할 때까지 무인 동물을 두지 마십시오. </li><li> 깨끗한 케이지로 다시 동물을 반환하기 전에 복구 할 수 있습니다. 완전히 회복 될 때까지 다른 동물의 회사에 동물을 반환하지 않습니다. </li><li> 관찰하고 매일 쥐를 무게. 쥐가 고통의 흔적이 보일 경우, 복강 1 멜 록시 캄로 주입 - 24 시간 - 2 밀리그램 / 각 12 kg이다. </li></ol> 8. 반복 매복 <ol><li> 일 4, 7 쥐에게 추가 부상을주고, 초기 손상 (충격 사이에 72 시간 간격) 이후 10. </li></ol> 9. 면역 조직 화학 (IHC) <ol><li> Transcardial 관류 <ol><li> 200m / kg 펜토 바르 비탈로 복강 내 주사를 통해 마우스를 마취. </li><li> 평가 및 발가락 핀치에 의해 수술 평면 마취를 확신합니다. 비서URE 부드럽게 화학 물질 흄 후드 내부에 스티로폼 작업 표면에 앞발과 뒷발을 테이핑으로 누운 자세에서 마우스. </li><li> 다만 쇄골에 칼 모양의 과정을 아래에서 흉부 중간 선을 따라 피부를 통해 절개를합니다. 칼 모양의 과정에서 두 개의 추가 피부 절개를 확인하고 옆으로 복부 흉곽의 기본을 따라 진행합니다. </li><li> 흉강을 열고 흉부 근육과 흉곽을 절단하여 심장을 노출. </li><li> 무딘 집게로 뛰는 심장을 보호하고는 1 - 좌심실에서 2mm 절개를. </li><li> 즉시 우심방에 나비 바늘을 삽입합니다. 천천히 주사기를 밀어 20ml의 식염수의 주입을 시작합니다. </li><li> 식염수에서 4 % 파라 포름 알데히드로 전환합니다. 파라 포름 알데히드의 20 ㎖와 재관류를 계속합니다. </li><li> 마우스를 목을 벨과 가위로 피부를 제거합니다. 뼈 커터를 이용하여 두개골로부터 뇌를 분리. </li></ol></li><li> 기음절편 ryostat <ol><li> -80 ° C에서 최적의 절삭 온도 OCT () 수립 및 동결에 포함 된 뇌 조직. 시상 방향으로 그라 이오 스탯의 뇌를 놓습니다. 잘라 뇌 섹션 5 μm의 두께. </li></ol></li><li> 더럽히는 것 <ol><li> 실온에서 1 시간 동안 고정 된 부분을 건조. </li><li> RT에서 1 시간 동안 인산염 완충 식염수 100 2 % 염소 혈청 μL, 0.1 % 트리톤 X-100 (PBS)로 슬라이드를 부화. </li><li> PBS 300 μL와 슬라이드 3 회 반복한다. 4 ℃에서 별도로 하룻밤 : (200 일) 또는 항 페리틴 항체 : 다음 안티 GFAP (200 일)와 슬라이드를 품어. </li><li> PBS 300 μL와 슬라이드 3 회 반복한다. 이어서 바이오틴 - 컨쥬 게이트 화 2 차 항체와 함께 실온에서 2 시간 동안 슬라이드를 배양한다. </li><li> PBS 300 μL와 슬라이드 3 회 반복한다. 이어서 실온에서 30 분 동안 아비딘 - 바이오틴 복합체 (ABC) 용액 (1시 50분)로 슬라이드를 배양한다. </li><li>PBS 300 μL와 슬라이드 3 회 반복한다. 8 분 - 그럼 3,3&#39;- 디아 미노 벤지딘 5 (DAB) 기질 용액 (50 ㎖ PBS, 10 μL의 H 2 O 2, 10 mg을 DAB 알약, 사용하기 전에 필터)에 품어. 긍정적 인 세포가 나타날 때까지 현미경 슬라이드를 관찰한다. </li><li> 5 분 동안 천천히 실행 수돗물에 슬라이드를 씻어. 실험실 닦아 청소 슬라이드. 그런 다음 설치 매체와 coverslip에와 섹션을 탑재합니다. </li></ol></li></ol>

<ol>
	<li>Baugh, C. 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=Chronic+traumatic+encephalopathy:+neurodegeneration+following+repetitive+concussive+and+subconcussive+brain+trauma.">Chronic traumatic encephalopathy: neurodegeneration following repetitive concussive and subconcussive brain trauma.</a> Brain Imaging Behav. 6 (2), 244-254 (2012).</li><li>McKee, A. 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=Chronic+traumatic+encephalopathy+in+athletes:+progressive+tauopathy+after+repetitive+head+injury.">Chronic traumatic encephalopathy in athletes: progressive tauopathy after repetitive head injury.</a> J. Neuropathol Exp Neurol. 68 (7), 709-735 (2009).</li><li>Petraglia, A. L., Dashnaw, M. L., Turner, R. C., Bailes, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Models+of+mild+traumatic+brain+injury:+translation+of+physiological+and+anatomic+injury.">Models of mild traumatic brain injury: translation of physiological and anatomic injury.</a> Neurosurgery. 75 Suppl (4), S34-S49 (2014).</li><li>Goldstein, L. E., McKee, A. C., Stanton, P. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Considerations+for+animal+models+of+blast-related+traumatic+brain+injury+and+chronic+traumatic+encephalopathy.">Considerations for animal models of blast-related traumatic brain injury and chronic traumatic encephalopathy.</a> Alzheimers Res Ther. 6 (5), 64 (2014).</li><li>Gold, E. 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=Functional+assessment+of+long-term+deficits+in+rodent+models+of+traumatic+brain+injury.">Functional assessment of long-term deficits in rodent models of traumatic brain injury.</a> RegenMed. 8 (4), 483-516 (2013).</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> Nat Rev Neurosci. 14 (2), 128-142 (2013).</li><li>Luo, 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=Long-term+cognitive+impairments+and+pathological+alterations+in+a+mouse+model+of+repetitive+mild+traumatic+brain+injury.">Long-term cognitive impairments and pathological alterations in a mouse model of repetitive mild traumatic brain injury.</a> Front Neurol. , 5-12 (2014).</li><li>Yang, Z., 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=Temporal+MRI+characterization,+neurobiochemical+and+neurobehavioral+changes+in+a+mouse+repetitive+concussive+head+injury+model.">Temporal MRI characterization, neurobiochemical and neurobehavioral changes in a mouse repetitive concussive head injury model.</a> Sci Rep. 10 (5), 11178 (2015).</li><li>Zhang, 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=Inhibition+of+monoacylglycerol+lipase+prevents+chronic+traumatic+encephalopathy-like+neuropathology+in+a+mouse+model+of+repetitive+mild+closed+head+injury.">Inhibition of monoacylglycerol lipase prevents chronic traumatic encephalopathy-like neuropathology in a mouse model of repetitive mild closed head injury.</a> J Cereb Blood Flow Metab. 35 (3), 443-453 (2015).</li><li>Petraglia, A. L., 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+spectrum+of+neurobehavioral+sequelae+after+repetitive+mild+traumatic+brain+injury:+a+novel+mouse+model+of+chronic+traumatic+encephalopathy.">The spectrum of neurobehavioral sequelae after repetitive mild traumatic brain injury: a novel mouse model of chronic traumatic encephalopathy.</a> J Neurotrauma. 31 (13), 1211-1224 (2014).</li><li>Lumpkins, K. M., Bochicchio, G. V., Keledjian, K., Simard, J. M., McCunn, M., Scalea, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glial+fibrillary+acidic+protein+is+highly+correlated+with+brain+injury.">Glial fibrillary acidic protein is highly correlated with brain injury.</a> J Trauma. 65 (4), 778-782 (2008).</li><li>Yang, Z., Wang, K. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glial+fibrillary+acidic+protein:+from+intermediate+filament+assembly+and+gliosis+to+neurobiomarker.">Glial fibrillary acidic protein: from intermediate filament assembly and gliosis to neurobiomarker.</a> Trends Neurosci. 38 (6), 364-374 (2015).</li><li>Liu, 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=Increased+expression+of+ferritin+in+cerebral+cortex+after+human+traumatic+brain+injury.">Increased expression of ferritin in cerebral cortex after human traumatic brain injury.</a> Neurol Sci. 34 (7), 1173-1180 (2013).</li><li>Jordan, B. D., 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+clinical+spectrum+of+sport-related+traumatic+brain+injury.">The clinical spectrum of sport-related traumatic brain injury.</a> Nat Rev Neurol. 9 (4), 222-230 (2013).</li></ol>

The authors have no financial interest to disclose.

임상 상태에 형태 학적으로 유사한 뇌 손상을 모방하기 위해 사후 뇌진탕 증상이 예상된다. 포스트 뇌진탕의 증상은 일반적으로 두통, 현기증, 현기증, 피로, 기억과 수면 문제, 문제가 집중뿐만 아니라 불안하고 우울한 기분을 포함한다. 신체 증상이 아직 동물 모델에서 측정되지 않을 수 있기 때문에, 모터 및인지 기능, 정서적 행동의 변화는 합리적으로 동물 모델에서 뇌진탕을 평가하기위한 ?...

뇌진탕은라고도 가벼운 외상성 뇌 손상 (mTBI), 외상성 뇌 손상 (TBI)의 가장 흔한 발생하고 미국에서 수백만의 사람에 영향을 미친다. 뇌진탕 진단하는 까다로운 일이 될 수 있으며, 충격에 대한 특별한 치료법은 없습니다. 성장 인식 및 스포츠 부상, 군사 전투 및 기타 물리적으로 결합 추구로 인한 경미한 기계적 외상 누적 만성 신경 학적 결과 1, 2를 가질 수 있다는 몇 가지 증거가있다. 그러나, 여전히 뇌진탕과 그 효과에 대한 지식의 부족입니다. 만 신경 학적 평가 및 이미징 평가는 임상 진단에 사용할 수 있기 때문에 현재 방법론은 인간의 병리와 치료의 연구를 제한합니다. 동물 모델은 추가 진단 및 mTBI의 치료의 희망 효율적이고 엄격하고 통제 된 방식으로 뇌진탕을 연구 할 수있는 수단을 제공한다. 연구는 기존의 TBI 적응이러한 제어 대뇌 피질의 영향 (CCI)와 같은 모델은 유체 타악기에 미치는 영향 (FPI), 체중 감소 부상 및 폭발 부상 mTBI을 수행하고 부상 매개 변수를 변경하여 낮은 부상의 심각도를 자극합니다. 이 모델 인해 임상 상태에 형태 학적으로 유사한 뇌 외상을 복제 할 수있는 능력을 사용하는 것이 유리하다; 그러나, 그들은 또한 그들 자신의 한계를 가지고있다. 가속 부상 (체중 감소)에 의해 유도 부상의 심각도는 종종 매우 가변적이다. 온화한 CCI의 두 가지 결과 - 지주막 하 출혈 및 초점 타박상은 - 전형적인 인간의 뇌진탕과 비교할 수 없습니다. 폭발 부상이 노출 3-6 동안 서로 다른 노출 위치 및 피크 압력 측정에 관해서 논쟁 모델뿐만 아니라 변수 보조 부상 상태에서 CCI와 FPI는 임상 적으로 관련없는 개두술을 필요로합니다. 업데이트 진탕 동물 모델을 즉, 임상 setti에 전임상 연구를 번역 할 수 있습니다NG 연구에 필요하다. 가벼운 TBI 모델링의 핵심 문제는 가장 밀접하게 임상 설정에서 부상을 복제 실험 부상의 심각도를 정의하는 것입니다. 최근 다른 연구 그룹은 폐쇄 머리 부상 또는 진탕 머리 부상 (CHI) 모델 7-10을 개발했다. CHI는 개두술없이 CCI의 변형이지만, 여전히 두부 충돌을 생성하는 기존의 전자파 영향 시스템을 사용한다. 카이는 효과 매개 변수를 조정하여 중간에 온화한에 이르기까지 뇌진탕을 유도 할 수 있습니다. 의식 (LOC)의 손실은 호흡 속도 또는 호흡 과도 종단의 감소를 검출함으로써 충격 후 즉시 관찰 될 수있다. LOC의 기간은 상처의 정도를 결정하는 데 사용된다. 본 논문은 자세한 단계별 프로토콜과 대표적인 결과와 함께 마우스의 반복적 인 CHI (rCHI) 모델의 약간 개선 및 업데이트 된 버전이 포함되어 있습니다. rCHI 모델 연구 전략 a를뇌진탕에 의한 병리학 적 변화를 모두 모방 할 수없는 각각의 동물 모델이 없습니다 특히 때문에, mTBI 효과와 잠재적 인 치료를 결정하는데 도움이 다시.

<table><tbody><tr><td>anesthesia machine</td><td>Eagle Eye Anesthesia, Inc</td><td>Model 150&amp;nbsp;</td><td>anesthesia</td></tr><tr><td>Electromagnetic Impactor</td><td>LeicaBiosystems</td><td>Impact One Stereotaxic Impactor</td><td>perform impaction</td></tr><tr><td>Digital Stereotaxic instrument</td><td>LeicaBiosystems</td><td>39462501</td><td>mount mouse and positioning tips</td></tr><tr><td>Sicilone rubber-coated metal tip</td><td>Precision Tool &amp;amp; Engineering, Gainesvill FL</td><td>custom-made</td><td>impact tip</td></tr><tr><td>Lithium Ion All-in-One Trimmer</td><td>WAHL Home Products</td><td>9854-600</td><td>shave mouse hair</td></tr><tr><td>paper clips</td><td>custom-made</td><td></td><td>probe tip</td></tr><tr><td>Cotton tipped applicators</td><td>MEDLINE</td><td>MDS202055</td><td>scrub head with saline</td></tr><tr><td>Tissue Tek O.C.T.</td><td>ASKURA FINETEK USA INC</td><td>4583</td><td>tissue embedding</td></tr><tr><td>anti-GFAP</td><td>Dako</td><td>CA93013</td><td>antibody for IHC</td></tr><tr><td>anti Ferritin</td><td>Sigma</td><td>F6136</td><td>antibody for IHC</td></tr><tr><td>VECTASTAIN Elite ABC&amp;nbsp; kit</td><td>Vector laboratories</td><td>PK-6100</td><td>IHC detection system</td></tr><tr><td>Permount Mounting Medium</td><td>Fisher Scientific</td><td>SP15-100</td><td></td></tr><tr><td>Aperio XT ScanScope scanner</td><td>Leica Microsystems Inc,</td><td></td><td>slides scanning</td></tr><tr><td>Leica AutoStainer XL</td><td>Leica the pathology Company</td><td>ST2010</td><td>H&amp;amp;E staining</td></tr><tr><td>DAB&amp;nbsp;</td><td>sigma</td><td>D3939</td><td>IHC detection system</td></tr></tbody></table>

a repetitive concussive head injury model in mice

Despite the concussion/ mild traumatic brain injury (mTBI) being the most frequent occurrence of traumatic brain injury, there is still a lack of knowledge on the injury and its effects. To develop a better understanding of concussions, animals are often used because they provide a controlled, rigorous, and efficient model. Studies have adapted traditional animal models to perform mTBI to stimulate mild injury severity by changing the injury parameters. These models have been used because they can produce morphologically similar brain injuries to the clinical condition and provide a spectrum of injury severities. However, they are limited in their ability to present the identical features of injuries in patients. Using a traditional impact system, a repetitive concussive injury (rCHI) model can induce mild to moderate human-like concussion. The injury degree can be determined by measuring the period of loss of consciousness (LOC) with a sign of a transient termination of breathing. The rCHI model is beneficial to use for its accuracy and simplicity in determining mTBI effects and potential treatments.

이 모델 (그림 1 AC)에서, 모든게 완벽 할꺼야 얕은 호흡의 짧은 기간이 있었다. (의식) 의식의 손실은 호흡 속도 또는 정상적인 호흡을 다시 시작하기 전에 호흡의 일시적인 중단의 감소로 정의된다. 머리의 중심에 미치는 영향은 단기 무의식의 원인 (7.5 ± 4.7, ± 5.5, 7.8, ± 8.8 10.2, 개별적으로 각각의 영향에서 9.5 ± 8.0 초, 그림 1D). 마우스의 뇌에 미?...

Watch this Scientific Journal Video about A Repetitive Concussive Head Injury Model in Mice at JoVE.com

A Repetitive Concussive Head Injury Model in Mice

Concussion presents the most common type of traumatic brain injury. Therefore, a repetitive concussive animal model, which replicates the important features of an injury in patients, may provide a means to study concussion in a rigorous, controlled, and efficient manner.

마우스에 반복적 인 충격 포 헤드 손상 모델

Despite the concussion/ mild traumatic brain injury (mTBI) being the most frequent occurrence of traumatic brain injury, there is still a lack of ...

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11.8K 조회수.  University of Florida. Concussion presents the most common type of traumatic brain injury. Therefore, a repetitive concussive animal model, which replicates the important features of an injury in patients, may provide a means to study concussion in a rigorous, controlled, and efficient manner.

비디오: A Repetitive Concussive Head Injury Model in Mice

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.

측면 유체 타악기 : 마우스의 외상성 뇌 손상 모델

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

연구

신경과학

A Mouse Model of Single and Repetitive Mild Traumatic Brain Injury

Mild Traumatic Brain Injury (mTBI) can result in the acute loss of brain function, including a period of confusion, a loss of consciousness (LOC), focal neurological deficits and even amnesia. Athletes participating in contact sports are at high risk of exposure to large number of mTBIs. In terms of the level of injury in a sporting athlete, a mTBI is defined as a mild injury that does not cause gross pathological changes, but does cause short-term neurological deficits that are spontaneously resolved. Despite previous attempts to model mTBI in mice and rats, many have reported gross adverse effects including skull fractures, intracerebral bleeding, axonal injury and neuronal cell death. Herein, we describe our highly reproducible animal model of mTBI that reproduces clinically relevant symptoms. This model uses a custom made pneumatic impactor device to deliver a closed-head trauma. This impact is made under precise velocity and deformation parameters, creating a reliable and reproducible model to examine the mechanisms that contribute to effects of single or repetitive concussive mTBI.

Athletes absorb several hundred mild traumatic brain injuries (mTBI)/concussions every year; however, the consequence of these on the brain is poorly understood. Therefore, an animal model of single and repetitive mTBI that consistently replicates clinically relevant symptoms provides the means to advance the study of mTBI and concussion.

단일 및 반복적 인 경미한 외상성 뇌 손상의 마우스 모델

Mild Traumatic Brain Injury (mTBI) can result in the acute loss of brain function, including a period of confusion, a loss of consciousness (LOC), focal ...

의학

A Novel and Translational Rat Model of Concussion Combining Force and Rotation with In Vivo Cerebral Microdialysis

Persistent cognitive and motor symptoms are known consequences of concussions/mild traumatic brain injury (mTBIs) that can be partly attributable to altered neurotransmission. Indeed, cerebral microdialysis studies in rodents have demonstrated an excessive extracellular glutamate release in the hippocampus within the first 10 min following trauma. Microdialysis offers the clear advantage of in vivo neurotransmitter continuous sampling while not having to sacrifice the animal. In addition to the aforementioned technique, a closed head injury model that exerts rapid acceleration and deceleration of the head and torso is needed, as such a factor is not available in many other animal models. The Wayne State weight-drop model mimics this essential component of human craniocerebral trauma, allowing the induction of an impact on the head of an unrestrained rodent with a falling weight. Our novel and translational rat model combines cerebral microdialysis with the Wayne State weight-drop model to study, in lightly anesthetized and unrestrained adult rats, the acute changes in extracellular neurotransmitter levels following concussion. In this protocol, the microdialysis probe was inserted inside the hippocampus as region of interest, and was left inserted in the brain at impact. There is a high density of terminals and receptors in the hippocampus, making it a relevant region to document altered neurotransmission following concussion. When applied to adult Sprague-Dawley rats, our combined model induced increases in hippocampal extracellular glutamate concentrations within the first 10 min, consistent with the previously reported post-concussion symptomology. This combined weight-drop model provides a reliable tool for researchers to study early therapeutic responses to concussions in addition to repetitive brain injury, since this protocol induces a closed-head mild trauma.

Neurotransmitter alteration is a mechanism of neural dysfunction that occurs after concussion and contributes to the sometimes-catastrophic long-term consequences. This rat model combines microdialysis, allowing in vivo neurotransmitter quantification, with a weight-drop technique exerting rapid acceleration and deceleration of the head and torso, an important factor of human craniocerebral trauma.

생체 뇌 미세 투석과 힘과 회전을 결합 하는 뇌진탕의 소설 및 번역 쥐 모델

Persistent cognitive and motor symptoms are known consequences of concussions/mild traumatic brain injury (mTBIs) that can be partly attributable to ...

Murine Model of Controlled Cortical Impact for the Induction of Traumatic Brain Injury

The Centers for Disease Control and Injury Prevention estimate that almost 2 million people sustain a traumatic brain injury (TBI) every year in the United States. In fact, TBI is a contributing factor to over a third of all injury-related mortality. Nonetheless, the cellular and molecular mechanisms underlying the pathophysiology of TBI are poorly understood. Thus, preclinical models of TBI capable of replicating the injury mechanisms pertinent to TBI in human patients are a critical research need. The controlled cortical impact (CCI) model of TBI utilizes a mechanical device to directly impact the exposed cortex. While no model can full recapitulate the disparate injury patterns and heterogeneous nature of TBI in human patients, CCI is capable of inducing a wide range of clinically applicable TBI. Furthermore, CCI is easily standardized allowing investigators to compare results across experiments as well as across investigative groups. The following protocol is a detailed description of applying a severe CCI with a commercially available impacting device in a murine model of TBI.

Here we describe a protocol for the induction of murine traumatic brain injury via an open-head controlled cortical impact.

외상성 뇌 손상의 유도에 대한 제어 피질 영향의 뮤린 모델

The Centers for Disease Control and Injury Prevention estimate that almost 2 million people sustain a traumatic brain injury (TBI) every year in the ...

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.

설치류의 폐쇄 머리 경미한 외상성 뇌 손상의 전임상 평가를위한 저강도 폭발파 모델

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 ...

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.

생쥐의 경미한 외상성 뇌 손상에 대한 전자기 제어 폐쇄 머리 모델

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

폐쇄형 머리 손상 방법을 통한 마우스의 rmTBI 유도

Modified Mouse Model of Repetitive Mild Traumatic Brain Injury Incorporating Thinned-Skull Window and Fluid Percussion

Mild traumatic brain injury is a clinically highly heterogeneous neurological disorder. Highly reproducible traumatic brain injury (TBI) animal models with well-defined pathologies are urgently needed for studying the mechanisms of neuropathology after mild TBI and testing therapeutics. Replicating the entire sequelae of TBI in animal models has proven to be a challenge. Therefore, the availability of multiple animal models of TBI is necessary to account for the diverse aspects and severities seen in TBI patients. CHI is one of the most common methods for fabricating rodent models of rmTBI. However, this method is susceptible to many factors, including the impact method used, the thickness and shape of the skull bone, animal apnea, and the type of head support and immobilization utilized. The aim of this protocol is to demonstrate a combination of the thinned-skull window and fluid percussion injury (FPI) methods to produce a precise mouse model of CHI-associated rmTBI. The primary objective of this protocol is to minimize factors that could impact the accuracy and consistency of CHI and FPI modeling, including skull bone thickness, shape, and head support. By utilizing a thinned-skull window method, potential inflammation due to craniotomy and FPI is minimized, resulting in an improved mouse model that replicates the clinical features observed in patients with mild TBI. Results from behavior and histological analysis using hematoxylin and eosin (HE) staining suggest that rmTBI can lead to a cumulative injury that produces changes in both behavior and gross morphology of the brain. Overall, the modified CHI-associated rmTBI presents a useful tool for researchers to explore the underlying mechanisms that contribute to focal and diffuse pathophysiological changes in rmTBI.

저자 스포트라이트: 반복적인 경미한 외상성 뇌 손상의 메커니즘 및 재활을 조사하기 위한 설치류 모델 최적화

This protocol presents a modified mouse model of repetitive mild traumatic brain injury (rmTBI) induced via a closed-head injury (CHI) method. The approach features a thinned-skull window and fluid percussion to reduce the inflammation commonly caused by meninges exposure, along with improved reproducibility and accuracy in modeling rmTBI in rodents.

얇아진 두개골 창과 유체 타악기를 통합한 반복적인 경미한 외상성 뇌 손상의 수정된 마우스 모델

Mild traumatic brain injury is a clinically highly heterogeneous neurological disorder. Highly reproducible traumatic brain injury (TBI) animal models ...

<meta charset="utf8"></head><body>CHILD 모델 — 생쥐의 청소년 뇌진탕 및 장기 결과 요약

A Pediatric Concussion Model in Mice: Closed Head Injury with Long-Term Disorders (CHILD)

Concussions and mild traumatic brain injuries (mTBI) during childhood represent a significant health endangerment, with many patients later in life exhibiting debilitating physiological, neurological, and psychosocial outcomes. The cellular and molecular pathophysiological mechanisms are relatively unknown, and this significant gap effectively precludes investigations into specific therapeutic strategies to mitigate chronic sequelae. No single animal concussion model recapitulates all the features reported in human subjects, and current models are designed to answer specific research questions. We set out to develop a juvenile concussion that recapitulates clinical early, and long-term symptomology encountered, termed Closed Head Injury with Long-term Disorder (CHILD). In this model, a concussive force via an electromagnetic impactor is delivered to the head of a lightly anesthetized unrestrained post-natal day 17 (P17) mouse, allowing free rotation of the head. Mice are placed on a tightly stretched tin foil across a stereotactic device, and the impactor is carefully aligned with the targeted cortical region. The electromagnetic impactor facilitates the selection of impact depth, dwell, and duration to determine injury severity. A strength of this model is that it allows head rotation, an important clinical feature. After impact, mice are immediately monitored for righting time and time to explore their environment, followed by their return to their dam. Increasing the severity of concussion results in intracranial bleeds in a subset of mice. Mice can be routinely monitored for behavior and neuroimaging over their lifespan, as desired. Various injury severities mimic the heterogeneous nature of juvenile concussions. The current standard CHILD model exhibits no skull fracture, no observable conventional neuroimaging change (similar to clinical), but leads to progressive and persistent neuronal death, altered diffusion MRI, modified neuronal activity and plasticity, increased gliosis, and progressive behavioral perturbations with age. In summary, this CHILD model mimics the early and long-term features observed in many clinical concussion patients.

<meta charset="utf8"></head><body>생쥐의 소아 뇌진탕 모델: 장기 장애를 동반한 폐쇄성 두부 손상(CHILD)

Pediatric concussions can result in long-term sequelae with physiological and psychological perturbations. This pediatric concussion mouse model results in long-term neuroinflammation, white matter alterations, and neuronal modifications with behavioral dysfunction. The model can be adapted to differential impact locations or repeated concussions to assess lifetime changes.

생쥐의 소아 뇌진탕 모델: 장기 장애를 동반한 폐쇄성 두부 손상(CHILD) 

Concussions and mild traumatic brain injuries (mTBI) during childhood represent a significant health endangerment, with many patients later in life ...

마우스에 반복적 인 충격 포 헤드 손상 모델

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