이 작품은 건강 보조금, R01 AG033620 (BG)의 국립 연구소에 의해 로버트 드레이퍼 기술 혁신 자금 (DAW)에 의해 지원되었다.

히트 장치 1. 건설 <ol><li> 두 클램프와 4 개의 나사 (그림 1A)를 사용하여 보드에 스프링을 부착합니다. 보드의 폭을 기준으로 클램프를 중심 보드의 가장자리 외부 클램프 플러시 서로에 대해 그들을 엉덩이. 이전에 클램프를 부착로, 봄을 통해 단단히 맞게 플라이어를 사용하여 구부. 주 : HIT 장치를 구성하는 데 필요한 자료에 대한 설명은 표 1을 참조하십시오. 스프링의 고정 단부는 기판의 가장자리로부터 1/8 인치 (3.2 mm)이어야하고, 자유 단부가 3/4 인치 (19mm)에 의해 기판 ​​위로 연장한다. 이 보드의 길이와 평행하게 놓 이도록 스프링을 조정합니다. </li><li> 벨크로 루프 접착 스트립 한번 주위 스프링의 자유 단부를 감싸. 벨크로의 바깥 쪽 가장자리는 봄의 끝과 접하게한다. 는 CRE에 의해 스프링 바이알을 고정하는 데 사용되기 때문에 중요하다 벨크로꽉 압축 피팅 체제상에서. 벨크로는 또한 많은 바이알 단시간에 처리 할 수​​ 있도록 쉽게 연결하고 튜브의 제거를 허용한다. </li><li> 단단한 나무 보드에 대해, 중심, 거꾸로 얼음 통 덮개를 놓습니다. 긴 쪽이 기판의 폭과 평행이되도​​록 얼음 통 덮개의 융 기부의 방향. 유리 병이 봄에 연결되어있는 경우 스프링 보드에 평평 거짓말을하지 않도록 얼음 통의 발생 지역, 나무 보드에 비해 1/2 인치 (13mm)보다 높은 있음을 유의하십시오. </li><li> 얼음 양동이 커버 보드와 객체 사이에 끼여하고 이동하지 않습니다 그래서, 벽과 같은 고정 된 개체에 대한 전체 장치를 밀어 넣습니다. </li><li> 테이프 판지의 바닥에 종이 각도기는 유리 병 트레이를 비행이 완벽하게 수직 위치로 철수 할 때 90 ° 마크가 봄과 정렬되도록 보드의 길이에 대해 가장자리에 서. </li></ol> 2.히트 장치의 작동 <ol><li> 빈 유리 병에 날아와 꽉 끼는면 공을 사용하여 병을 마개 -anesthetized 1과 60 CO 2 사이에. </li><li> 이 바닥에서 1 인치 (2.5 cm) 때까지 병에 목화 공을 밀어 유리 병의 바닥 1 인치 (2.5 cm)에 파리를 제한 할 것. 그것은 1 인치 (2.5 cm) 시점에서 유리 병에 선을 그어야하는 것이 도움이된다. 유리 병의 더 크거나 작은 지역에 파리를 구속하는 것은 표현형의 정도에 영향을 미칠 수 있습니다. </li><li> 파리는 CO 2에서 이동성을 복구하기 위해 5 분을 기다립니다. 이 5 분을 완전히 CO 2의 효과를 제거 할 수있는 여부를 공지되지 않는다. </li><li> 벨크로의 내부 가장자리가 유리 병 (그림 1B)의 상단과 수평이 될 때까지 유리 병에 봄의 끝을 삽입합니다. 봄이 평평 거짓말을하면, 유리 병의 1 인치 (2.5 cm)는 얼음 양동이 커버의 발생 영역을 중첩한다. 유리 병은 M을 재사용 할 수 있습니다모든 시간. </li><li> 앉아있는 동안, 왼손의 엄지와 집게 손가락을 사용하여 벨크로 지역에서 병을 개최합니다. 오른손을 사용하여 벤치 탑에 보드가 꽉 잡아. 또한, 벤치 탑에 꽉 이사회를 개최 C - 클램프를 사용합니다. </li><li> 완벽하게 직선 다시 원하는 각도에 스프링을 당깁니다. 봄을 놓습니다. 봄이 완전히 정지 할 수 있도록합니다. </li><li> 봄의 병을 제거하고 파리 ≥5 분 동안 복구 할 수 있습니다. 다른 파업으로 파리를 주제 또는 플라이 음식과 함께 유리 병에 파리를 전송합니다. 주 : 검정법의 다양한 HIT 장치로부터 공격의 표현형 효과를 평가할 수있다. 예를 들어, 수명에 영향이 손상 후 항상 생존 파리의 비율을 분석함으로써 결정될 수있다 뇌 형태에 미치는 영향은 헤드의 조직 학적 분석에 의해 측정 될 수 있고, 유전자 발현에 대한 효과는 mRNA 수준의 정량 분석​​에 의해 결정될 수있다 10. </li><li> Determin동일 파업을 실시하지 않는 제어 파리 처리하여 파업으로 인해하지 않은 절차의 전자 효과. 얼음 양동이 커버에 대한 유리 병의 영향이 큰 소음이 발생하기 때문에 청력 보호 착용 할 것. </li></ol>

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

우리는 공개 관심 없음 충돌이 없습니다.

히트 장치 방법은 TBI (11)을 관통보다는 폐쇄 머리를 유발한다는 사실에 의해 파리에서 외상을 입힐 다른 방법과 구별된다. 또한, HIT 장치 방법은 많은 초파리 TBI를 입힐 적은 시간, 노력과 기술을 걸리므 방법은 대규모의 유전자 스크린에 대한 다른 방법들보다 더 의무이다. 마지막으로, HIT 디바이스에 의해 가해진 프라이 머리 부상에 한정되지 않는다는 사실은 한계와 이점을 모두이다. ?...

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

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

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

외상성 뇌 손상 (TBI)가 외부로부터 기계적인 힘 뇌 손상으로서 정의된다. 가장 일반적으로, 같은 무딘 힘과 관성 가속과 뇌가 두개골의 내부를 공격하게 감속 힘으로 닫힌 헤드 세력 TBI 결과. 미국에서, 5 만 개인이 TBI 매년 죽고 2.5-6500000 개인, TBI의 결과와 함께 살고 물리적,인지 쇠약 포함, 1,2- 행동 문제가있는 것으로 추정된다. TBI의 결과로 인해 뇌에 차 기계 부상뿐만 아니라 시간이 3-5에 걸쳐 발생하는 뇌에 보조 세포 및 분자 부상뿐만 아니라 다른 조직에뿐만 아니라입니다. TBI 진단 및 치료 방법의 개발은 TBI는 복잡한 질병 과정이므로 어려운 것으로 입증되었다. 주 부상, 인간 생리학, 환경 요인의 변수 특성은 이기종 보조 나는 결과njuries 및 병리. 근본적인 요인 변수는 개인의 주요 손상의 중증도, 반복적 차 부상 사이의 시간, 및 연령 및 유전자형을 포함한다. 각 변수 요인이 TBI의 결과에 기여하는 방법을 이해하는 것은 진단하고 TBI 6,7 치료 방법의 개발에 도움을 가능성이 높습니다. 여기에서 우리는 TBI의 결과에 변수 요인의 기여를 묘사하는 데 사용할 수있는 초파리 (초파리)에 폐쇄 머리 TBI를 입힌하는 방법을 설명합니다. 방법은 강렬하게 손의 손바닥에 비행 문화 유리 병의 측면을 타격하는 야생형 발생 초기 관찰을 기반으로 일시적, TBI의 가능성이 결과를 무력화되기 위해 파리. 따라서, 우리는 손으로 치는 동작에서 가속 및 감속 힘을 요점을 되풀이하는 높은 충격 외상 (HIT) 장치를 건설했다. 고속 동영상 보여준다에서 단일 스트라이크장치가 원인 HIT는 머리와 몸 8 유리 병 벽을 여러 번 연락을 날아. 어느 정도, 모든 연락처는 총알과 폭포와 자동차 충돌 9 인간에게 발생하는 것과 유사한 머리 캡슐에 대해 변형 플라이 뇌가 발생할 가능성이 있습니다. 따라서, 뇌 10 실조증이어서 임시 무능력 이동성의 점진적인 회복, 헤드에서의 유전자 발현의 변화 및 진행성 신경 퇴행을 포함한 뇌 손상과 일치 HIT 장치 디스플레이 표현형 처리 날아간다. 따라서, HIT 장치는 가능한 실험 도구와 파리 위해 개발 된 기술을 이용하여 엄청난 아스날 TBI를 연구 할 수있다.

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

a method to inflict closed head traumatic brain injury in drosophila

외상성 뇌 손상 (TBI)는 물리적,인지 적, 행동 적 기능과 죽음의 손상을 초래, 매년 수백만의 사람에 영향을 미친다. 초파리를 이용한 연구는 신경 학적 과정을 이해에서 중요한 돌파구를 기여했다. 따라서, 인간의 TBI 병리의 세포 및 분자 기초를 이해하는 것을 목표로, 우리는 파리에 폐쇄 머리 외상성 뇌 손상을 입힐 수있는 높은 충격 외상 (HIT) 장치를 개발했다. 파리 임시 무능력 및 신경 퇴화와 같은 진행성 인간 TBI 부합 HIT 장치 디스플레이 표현형을 실시. HIT 장치는 플라이 뇌에 기계적인 손상을 초래, 유리 병의 벽에 파리를 추진하는 스프링 기반 메커니즘을 사용합니다. 장치는 자신의 조작이 간단하고 빠르며, 시공이 저렴하고 용이하며, 또한 재현 가능한 결과를 생성한다. 따라서, HIT 장치는 기존의 실험 도구와 파리는 근본적인 해결하기위한 기술과 결합 될 수있다외상성 뇌 손상에 대한 진단 및 치료의 발전으로 이어질 수 TBI에 대한 질문. 특히, HIT 장치 TBI 병리의 유전 기초를 이해하기 위해 대규모의 유전자 스크린을 수행하는데 사용될 수있다.

우리는 파리가 차 손상 후 곧 죽을 이유를 이해하는 데 관심이 있습니다. 죽음을 정량화하기 위해, 우리는 차 부상의 24 시간 이내에 사망 파리의 비율을 24 시간에서 사망 지수 (MI 24)를 결정했다. 히트 장치에서 파업을 실시 파리는 플라이 식품 유리 병에 25 ℃에서 배양하고, 죽은 파리의 수는 24 시간 후에 계산되었다. 우리는 MI (24)에 영향을 미치는 요소를 식별하기 위해이 방법을...

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A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila

여기에서 우리는 초파리에서 폐쇄 머리 외상성 뇌 손상 (TBI)를 입힐 수있는 방법을 설명합니다. 이 방법은 파리에 사용할 실험 도구와 기술의 광대 한 배열을 사용 TBI 병리의 기초 세포와 분자 메커니즘을 조사하기 위해 게이트웨이를 제공합니다.

폐쇄 머리 외상성 뇌 손상을 입힐 수있는 방법<em&gt; 초파리</em

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

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Neuroscience

A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila

11.4K 조회수.  University of Wisconsin-Madison.  여기에서 우리는 초파리에서 폐쇄 머리 외상성 뇌 손상 (TBI)를 입힐 수있는 방법을 설명합니다. 이 방법은 파리에 사용할 실험 도구와 기술의 광대 한 배열을 사용 TBI 병리의 기초 세포와 분자 메커니즘을 조사하기 위해 게이트웨이를 제공합니다. 

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

Lateral Fluid Percussion: Model of Traumatic Brain Injury in Mice

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

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

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

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

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

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

가벼운 외상성 뇌 손상 후 Hippocampal 회로 기능의 수선에 관한 조사

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

A Novel Method to Model Chronic Traumatic Encephalopathy in Drosophila

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

A Novel Method to Model Chronic Traumatic Encephalopathy in Drosophila

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

만성 외상성 뇌증 모델링을위한 새로운 방법<em&gt; Drosophila</em

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

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

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

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

외상 성 부상에 의해 영향을 하는 뇌 영역의 정위 적 아틀라스 유도 레이저 캡처 기정

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

A Novel In Vitro Model of Blast Traumatic Brain Injury

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

A Novel In Vitro Model of Blast Traumatic Brain Injury

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

모델을 새로운 시험관에 폭발 외상 성 뇌 손상의

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

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

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

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

가압된 설치류에 폭발을 이용한 Neurotrauma의 효과 중간 대뇌 동맥

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

Investigating Alterations in Caecum Microbiota After Traumatic Brain Injury in Mice

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

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

쥐에서 외상성 뇌 손상 후 Caecum Microbiota에서 변경 을 조사

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

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

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

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

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 ...

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

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

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

외상성 뇌 손상에 대한 신경 염증 및 혈역학 반응의 시스템 분석

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

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

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

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

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 ...