The authors would like to thank Irene Ma, Rose Tobais, and Jong Rho for their technical assistance. Funding was provided to MJE by the Department of Pediatrics at the University of Calgary, the Alberta Children&#8217;s Hospital Foundation (ACHF) and the Alberta Children&#8217;s Hospital Research Institute (ACHRI). The Postdoctoral fellowship for RM was provided by ACHF.

참고 : 모든 실험 동물 관리의 캐나다 협의회에 따라 수행하고, 캘거리 대학, 동물 보호 윤리위원회에 의해 승인되었다. 1. 사육 및 동물 준비 <ol><li> 표준 사육 프로토콜에 따라 사내 표준 실험 동물 공급 업체 또는 품종의 새끼에서 임신 한 쥐를 주문하십시오. </li><li> 어두운주기 : 하우스 온도 조절 쥐 주택 방 (21 ° C)에서 음식과 물에 대한 광고 무제한 액세스, 모든 동물, 즉 12시 12분 시간 빛에 유지됩니다. </li><li> 새끼는 생후 21 일 (P21)에 도달하면, 3 또는 4의 동성 그룹에서 자신의 어머니와 집에서 새끼를 유아. </li></ol> 가벼운 외상성 뇌 손상 (mTBI) 장치의 2. 업 <ol><li> 절차의 시작 이전에, 원하는 질량 밀 가중치 (예 150g). 안전하게 낚싯줄 Fi 접속 될 수 있도록 가중치의 상단에 금속을 부착 루프무게에 고정 된. </li><li> 날카로운 면도날 주석 호일 점수. 득점 주석 박 쥐의 체중을 지원하지만 무게와 헤드의 임팩트 후에 쥐의 가속을 방해하지 있는지 확인합니다. </li><li> 이 팽팽하게되도록, 컬렉션 스폰지 (38 X 25 X 15cm 3) (그림 1)을 포함 테이프 투명한 플라스틱 (38 X 27 X 27cm 3)로 만든 U 자형 무대에 득점 깡통 호. </li><li> 투명한 플라스틱으로 이루어지는 가이드 튜브 아래 올바른 위치에 U 자형 플라스틱 스테이지를 놓는다. </li><li> 클램프 스탠드 대신에 플라스틱 가이드 튜브 (2.2 cm 직경 X 1.5 m)를 잡고는 득점 주석 호일 위에 3.5 cm가되도록 가이드 튜브를 놓습니다. </li><li> 무게의 바닥이 자유롭게 2.5 cm 득점 주석 호일 위에 달려 보장 무게에 금속 루프를 통해 낚시 줄을 연결합니다. </li><li> 확실하게 클램프 스탠드에 낚시 줄을 연결합니다. 참고 : 조개에 무게를 테 더링쥐 컬렉션 스폰지에 미치는 영향)에서 떨어지는 동안 주석 호일 위에 2.5 cm의 높이에서 P 스탠드 다시 안타를 방지 할 수 있습니다. </li><li> 낚시 줄과 플라스틱 가이드 튜브를 통해 무게를 당겨 1.0 m에서 렌 키 핀 제자리에 고정. </li></ol> mTBI 3. 유도 <ol><li> 쥐 절차 방에 P30 이동 케이지에 도달하면. </li><li> 이소 플루 란 챔버에서 쥐를 놓고이 발이나 꼬리 핀치에 응답하지 않는 때까지 가볍게 쥐를 마취. </li><li> 빠르게 떨어지는 무게의 경로에서 직접 머리를 득점 주석 호일에 쥐의 가슴 아래로 놓습니다. 그림 2를 참조하십시오. 쥐를 이동하거나 웨이크 업이 득점 주석 호일에 배치되기 전에, 발 또는 꼬리 핀치에 응답하지 않는 때까지 이소 플루 란 챔버로 돌아가서 다시 시작하기 시작합니다. </li><li> 무게가 플라스틱 가이드 튜브를 수직으로 떨어져 머리에 쥐를 공격 할 수 있도록 육각 렌치 핀을 당기십시오. 목전자 쥐 빠르게 앙와위에서 180 ° 회전 및 땅을 받게 될 것이다. </li><li> 즉시 수집 스폰지에서 쥐를 제거하고면 팁 어플리케이터를 쥐의 머리에 국소 리도카인을 적용합니다. </li><li> 시중에서 판매하는 가열 패드에 설정하여 가열 깨끗한 케이지에서 앙와위에서 쥐를 놓습니다. </li><li> 스톱워치를 이용하거나 디지털 타이머 &quot;타임 투 오른쪽&quot;을 취득. 시간에서 오른쪽으로는 마취에서 깨어나 쉬운 위치로 앙와위에서 플립 또는 도보를 시작하기 위해 쥐에 의해 걸리는 시간입니다. </li><li> 이 정상적인 동작 (등, 탐구, 손질 산책을) 복구 후 홈 케이지에 쥐를 돌려줍니다. </li><li> 반복 mTBI을 필요로하는 각 추가 쥐에 대한 3.2-3.8 단계를 반복합니다. </li><li> 반복 mTBI 실험에 대해 여러 시점에서 동일한 쥐의 절차를 반복합니다. </li></ol> 위장 손상 4. 유도 <ol><li> 가볍게 ANEST이 발이나 꼬리 핀치에 응답하지 않는 때까지 이소 플루 란과 쥐를 hetize. </li><li> 빠르게 떨어지는 무게의 경로에서 직접 머리를 득점 주석 호일에 쥐의 가슴 아래로 놓습니다. </li><li> 플라스틱 가이드 튜브에서 렌 키 핀을 당기는없이 깡통 호에서 쥐를 제거하고면 팁 어플리케이터를 쥐의 머리에 국소 리도카인을 적용합니다. </li><li> 시중에서 판매하는 가열 패드에 설정하여 가열 깨끗한 케이지에서 앙와위에서 쥐를 놓습니다. </li><li> &quot;시간에서 오른쪽으로&quot;를 획득 스톱워치 또는 디지털 타이머를 사용합니다. </li><li> 이 정상적인 동작 (등, 탐구, 손질 산책을) 복구 후 홈 케이지에 쥐를 돌려줍니다. </li></ol> 빔 워킹 테스트 8 mTBI 5. 확인 <ol><li> 절차 방에 mTBI 또는 가짜 부상 복귀 동물의 유도 후 24 시간. </li><li> 깨끗한 유지 케이지에 쥐를 전송및 홈 케이지의 개방형 부분은 빔의 폭이 좁은 끝을 향하도록 빔을 테이퍼 165cm의 좁은 끝에서 쥐의 홈 케이지를 놓습니다. 참고 : 테이퍼 빔은 165cm 깁니다. 빔의 중앙 플랫폼은 광각 단에서 6cm 좁은 단부에서 1.75 cm의 폭을 가진다. 센터 빔이 때 쥐의 발 전표 안전을 제공 선반 (2cm 폭 중심 빔 아래 2cm)가 있습니다. 시험 중에 빔 추락 할 수 쥐에 부상의 위험을 줄이기 위해 빔 아래에 거품 패딩을 놓습니다. </li><li> 테이퍼 빔 위치의 광각 단에서 비디오 카메라를 배치 / 비디오 시청 조사자 명확 테이퍼 빔의 길이 전체 쥐의 움직임을 볼 수 있도록 비디오 카메라 확대. </li><li> 테이퍼 빔의 넓은 끝에서 쥐를 놓고 홈 케이지에 빔을 가로 질러 걸어을 권장합니다. 쥐가 테이퍼 빔을 통과하면, 레이 적어도 60 초 동안 홈 케이지에 쥐를 남겨대상 위치를 엔포스. 이것은 시험 # 1이고, 분석에 포함되지 않음. </li><li> 의 비디오 카메라를 켜고 쥐 빔 도보 작업을 4 번 더 추가 (홈 케이지에서 60 초 보강 기간을 유지)를 완료 할 수 있습니다. 쥐가 모든 시험을 완료하면, 홈 케이지에 쥐를 반환합니다. </li><li> 점수로 발생 시간 / 기간은 각각의 재판, 빔을 건너 뒷다리 발 - 전표의 수를 기록한다. 각 쥐​​ 빔을 건너 뒷다리 발 - 전표 및 시간의 평균 수를 계산하기 위해이 데이터를 사용합니다. </li></ol>

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No competing financial interests exist.

Reliable modelling systems are needed to effectively cultivate basic science research that has significant translational validity. In response to rising occurrences and popular media, the investigation of mTBI and concussion has become a priority in many disciplines. However, despite increased research, there have been only incremental improvements in therapeutic strategies and treatment options 3. This lack of progress may be partially due to a discrepancy between the modeling systems employed and actual injury etiology. The majority of studies utilized rodent models that failed to reproduce the important biomechanical forces and appropriate post-injury symptomology. The current human definition of mTBI specifies that the injury results from acceleration and deceleration forces associated with a blunt trauma 10. The modified weight drop technique described here is therefore an ideal model for the study of mTBI and concussion because it uses a glancing impact to cause rapid rotational acceleration and deceleration to the head of an unrestrained animal, mimicking the biomechanical forces identified in sports-related injuries and automobile accidents. In addition, this model is easily adapted to examine repetitive mTBI, a phenomena that is emerging as a serious medical and socioeconomic issue. Studies indicate that rodents may be exposed to a series of 10 distinct mTBIs with minimal mortality 7. Finally, the method is inexpensive and can be carried out rapidly, allowing for high-throughput examination of a many therapeutic compounds and treatment regiments. 
Just as with any procedural technique, certain aspects of the protocol are particularly important to the generation of reliable results. First, the tin foil needs to be scored effectively. If the tin foil is not properly scored, the force imparted by the weight during the glancing impact will not be enough to propel the juvenile rat through the tin foil onto the collection sponge. In these situations, the rat will remain in the starting position (chest down on the tin foil) and the mTBI will result from the blunt trauma from the weight impacting the stationary head, not the rotational acceleration and deceleration desired. Second, during the induction of the mTBI and the sham injury, the level of anesthetic applied to each rat should be consistent. Owing to the fact that time-to-right is used as marker of mTBI, the researcher should try to ensure that animals receiving a mTBI and animals receiving a sham injury are exposed to similar levels of anesthetic. A major advantage to this technique over many other TBI procedures is the low level and duration of anesthesiology. However, the juvenile rat needs to be non-responsive to a toe or tail pinch to ensure they do not wake-up on the stage before the injury is induced. Finally, in order to maintain a consistent injury etiology, the positioning of the rat&#8217;s head is particularly important. Ideally the weight should impact the center of the dorsal surface of the head. Caution should be taken to avoid positioning the path of the weight too near the caudal/posterior portion of the head, as impacting the brainstem and cerebellum is associated with increased mortality and seizure activity. 
Based upon the biomechanical pathophysiology of injury induction and the behavioural outcomes examined, the modified weight-drop technique appears to be a reliable model for the investigation of paediatric mTBI and concussion. Although preliminary studies of this novel model have assessed some basic molecular and structural changes 7 future studies will be needed to ascertain how the brain responds to a mTBI with this injury etiology. An in-depth analysis of the neuroanatomical and biological changes that occur at the cellular and epigenetic level would increase model validity and translational applicability. In addition to stimulating the generation of targeted pharmacological therapies, understanding the pathophysiological changes that occur in the brain in response to mTBI and concussion would also direct the research related to clinical biomarkers that have the ability to predict outcomes following injury.

중등도에서 중증 외상성 뇌 손상 (TBI), 거의 기술이 약한 유도하기 위해 개발되었습니다의 생성을위한 많은 널리 사용되는 방법이 있지만, 설치류에서 머리 부상을 마감했다. 경증 외상성 뇌 손상 (mTBI)가 세 배 더 일반적인 복합 1 중등도 뇌 손상보다 사실에 의해, mTBI의 안정적인 모델 연구에 대한 병태 생리, 신경 생물학 및 행동의 결과, 및 치료 전략을 용이하게하기 위해 필요하다. 예를 들어, 200 가지가 넘는다 지난 10 년간, 현재 동물 모델 2의 한계에 부분적으로는 TBI (3)의 치료를 위해 약물 임상 시험에 실패했습니다. 모델링 시스템이 병진 조사 연구를 위해 생성되는 경우, 결과의 적용 구현 모델의 유효성에 의존한다. mTBI / 뇌진탕의 연구를 위해 신뢰할 수있는 동물 모델은 생체 역학적 인 힘 respon을 모방하지 않을부상의 원인에 대한 sible뿐만 아니라 임상 적으로 인구에 의해보고 된 것과 일치 증상을 유발한다. 아이들이 mTBI에 특히 높은 위험이 있기 때문에 또한, 최적의 모델링 시스템은 성인 등가물뿐만 아니라, 젊은 청소년 설치류에 적용 할 수있다. 선수 mTBIs 또는 진탕 뇌 손상을 입었다 고하는 상황의 생체 역학적 분석은 부상의 가장 중요한 예측 요인 빠른 머리 가속 및 고속 영향 4 있음을 나타냅니다. 현재 TBI의 유도에 이용 설치류 모델의 대부분은 헤드 (5) (검토를 위해 2 참조)를 거의 또는 전혀 이동을 허용한다. 여기에 설명 된 모델은 피사체의 머리와 몸에 가속 / 감속의 힘을 적용 180 ° 회전 및 자유 낙하를 동반 물리적으로 억제되지 않은 청소년 쥐의 머리에 높은 속도에 미치는 영향을 제공합니다. 티여기에 mTBI의 유도에 대해이 수정 된 체중 감소 기술과 관련된 두 가지 주요 장점은. 첫째, 모델 (행동의 결과가 6 참조의 완전한 설명) 뇌에 대한 명백한 손상을 유발하지 않고 symptomology 같은 임상 적으로 진탕을 생산하고 있습니다. 후 진탕 증후군의 임상 보고서와도 일관성이 수정 체중 감소 기술은 이기종 결과를 생성합니다. mTBI의 효과가 중요하지만, 여러 결과 측정에 검사 할 때 mTBI을 경험 설치류 사이에 상당한 차이가 있습니다. 둘째, 본 방법은 반복 mTBI (7)의 조사를 허용한다. 기존 TBI 대부분 모델은 심각한 상해를 입힐 때, 그것은 전체 피질 광범위한 손상없이 반복적 TBI 공부 종종 초 부상을 충분히하는 것이 곤란하고, 거의 불가능하다. 따라서, 산업용 냉장고 변성 중량 드롭 기술을 이용하기위한 기본 원리mTBI의 ction의 더 밀접 청소년 인구의 병태 생리 및 뇌진탕의 symptomology 반복적 TBI를 나타내는 부상을 생산하는 것입니다. 특히 어린 시절 동안 증가 스포츠 관련 mTBI의 발생, 폭포, 자동차 사고로, mTBI의 독특한 설치류 모델은 쉽게 여러 히트에 적용 할 수 진탕 같은 뇌 손상의 연구를위한 유용한 도구와 연구자를 제공합니다 패러다임.

<table border="0" cellpadding="0" cellspacing="0" width="824">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:264px;">Brass Weights</td>
			<td style="width:155px;">Ginsberg Scientific</td>
			<td style="width:187px;">7-2500-2</td>
			<td style="width:219px;">Need to have metal loop attached to base</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:264px;">Alluminum Foil</td>
			<td style="width:155px;">Alcan</td>
			<td style="width:187px;"></td>
			<td>Available at most grocery stores</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:264px;">Masking Tape</td>
			<td style="width:155px;"></td>
			<td style="width:187px;"></td>
			<td>Commercially available&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:264px;">U-Shaped Plastic Stand</td>
			<td style="width:155px;"></td>
			<td style="width:187px;"></td>
			<td>Constructed by Laboratory</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:264px;">Clamp Stand</td>
			<td style="width:155px;">Sigma-Aldrich</td>
			<td style="width:187px;">Z190357</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:264px;">Plastic Guide Tube</td>
			<td style="width:155px;"></td>
			<td style="width:187px;"></td>
			<td>Could be constructed or purchased at a hardware store</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:264px;">Fishing Line</td>
			<td style="width:155px;">Angler 10lb&#160;</td>
			<td style="width:187px;"></td>
			<td>Purchased from a sporting goods retailer&#160;</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:264px;">Isoflurane</td>
			<td style="width:155px;">Pharmaceutical Partners of Canada</td>
			<td style="width:187px;">DIN 02237518</td>
			<td>Inhalation Anesthetic</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:264px;">Topical Lidocaine (30ml)</td>
			<td style="width:155px;">Astra Zeneca</td>
			<td style="width:187px;">DIN 0001694</td>
			<td>Xylocaine Jelly 2%&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:264px;">Cotton Swabs</td>
			<td style="width:155px;"></td>
			<td style="width:187px;"></td>
			<td>Commercially available&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:264px;">Heating Pad - 3 heat setting</td>
			<td style="width:155px;"></td>
			<td style="width:187px;"></td>
			<td>Commercially available&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:264px;">Stop Watch</td>
			<td style="width:155px;">Sportline</td>
			<td style="width:187px;">L303</td>
			<td>Purchased from a sporting goods retailer&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:264px;">Video Camera</td>
			<td style="width:155px;">Sony</td>
			<td style="width:187px;">HDR-CX260V</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:264px;">Sprague Dawley Rats</td>
			<td style="width:155px;">Charles River Laboratories</td>
			<td style="width:187px;">SAS SD 40</td>
			<td>Male and females ordered from Charles River Laboratories and pups bred in-house</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:264px;">Balance Beam</td>
			<td></td>
			<td></td>
			<td>Constructed by Laboratory</td>
		</tr>
	</tbody>
</table>

a novel model of mild traumatic brain injury for juvenile rats

Despite growing evidence that childhood represents a major risk period for mild traumatic brain injury (mTBI) from sports-related concussions, motor vehicle accidents, and falls, a reliable animal model of mTBI had previously not been developed for this important aspect of development. The modified weight-drop technique employs a glancing impact to the head of a freely moving rodent transmitting acceleration, deceleration, and rotational forces upon the brain. When applied to juvenile rats, this modified weight-drop technique induced clinically relevant behavioural outcomes that were representative of post-concussion symptomology. The technique is a rapidly applied procedure with an extremely low mortality rate, rendering it ideal for high-throughput studies of therapeutics. In addition, because the procedure involves a mild injury to a closed head, it can easily be used for studies of repetitive brain injury. Owing to the simplistic nature of this technique, and the clinically relevant biomechanics of the injury pathophysiology, the modified weight-drop technique provides researchers with a reliable model of mTBI that can be used in a wide variety of behavioural, molecular, and genetic studies.

상기 개질 중량 놓기 기법 청소년 래트에서 경증 외상성 뇌 손상 (mTBI)의 안정적인 유도 방법이다. 150g의 중량 충격을 살린,이 기술은 성공적 50~120g의 범위 청소년 래트에 적용되어왔다. 또한, 반복적 인 절차를 쉽게 mTBI의 연구를위한 동물에서 동일한 반복 될 수있다. (도 3) 단일 mTBI 전시에게 타임 투 오른쪽의 증가를 경험하고 나타나는 동물 기절 깨우면 있지만, 그들은 빠르게 정상적인 활동을 재개하고 가성 동물 부상 시각적으로 구별 할 수 없다. 부상이기는 영향과 관련된 통증을 제거, 국소 리도카인이 필요한 유일한 진통이 가벼운입니다 감안할. 진통제는 일반적인 염증 및 복구 프로세스를 방해하는 것으로 알려져있다 이것은 연구에 중요하다. 명백한 symptomology의 부족으로 인해, 빔 보행 태스크에 안정mTBI의 유도를 확인하는 데 사용할 수 있습니다 OL. 그것은 mTBI가 발생하지 모든 동물은 빔 산책 작업에 적자를 나​​타낼 것으로,주의하는 것이 중요하지만, 가짜 부상으로 청소년 쥐에 비해 그룹으로, mTBI와 청소년 쥐가 훨씬 더 뒷다리 발 - 슬립을 보여 (그림 4). 이 변형 중량 놓기 기술의 또 다른 중요한 특징은 부상 유도시 청소년 래트인가 구속의 부족이다. 빠른 회전 가속과 감속 다음에 머리에 반짝이기는 타격을 제공함으로써,이 모델은 더 밀접하게 mTBI 및 진동에 의한 생체 역학적 힘을 나타냅니다. 이 절차가 청소년 쥐 또는 성인 마우스에 적용하면, 사망률 (2백2분의 7 청소년 동물 ~ 3.4 %의 사망률) 매우 낮은이며, 두개골 골절 및 두개 내 출혈은 6,7 매우 드물다. 또한, 임상 적으로 적절한 모델을 생성 SYmptomology. 단일 mTBI을 경험 청소년 설치류는 우울증 같은 행동 증가 집행 기능 적자, 및 변경 사회적 상호 작용 6,9과 함께 균형과 모터 행동에 적자를 보여 주었다. 마찬가지로, 성인 마우스는 또한 시간 7 복구 온화한 균형과 조정 적자를 표시합니다. 마지막으로, mTBI이 모델을 사용하여 유도는 최소한의 마취가 필요하고 수술 준비 또는 두개골에내는 포함되지 않습니다. 결과는 따라서 수술이나 마취에 의해 트리거 혼란 염증 또는 면역 학적 효과에 의해 바이어스되지 않습니다. 또한, 열려있는 상처의 빠른 회복 시간과 부족은 설치류가 mTBI을 경험 한 직후 발생하는 추가 테스트 패러다임의 시작을 할 수 있습니다. <img alt="그림 1" fo:content-width="5in" src="/files/ftp_upload/51820/51820fig1highres.jpg" width="500" /> 그림 1 : </strong<html> &gt; 모든 관련 치수 플라스틱 단계 및 수집 스폰지 U 자형. 10cm의 거리가 수집 스폰지와 청소년 쥐를 보장하는 플라스틱 스테이지의 상부 사이에 유지되어야의 C의 artoon 표현 (180)을 완료하기에 충분한 시간을 갖는다 ° 회전. <img alt="그림 2" fo:content-width="5in" src="/files/ftp_upload/51820/51820fig2highres.jpg" width="500" /> 그림 2 : 부상 유도 플랫폼의 (A) 사진 표현입니다. 머리가 떨어지는 무게 바로 아래에 있도록 청소년 쥐 득점 주석 호일에 가슴이 아래로 배치됩니다. 부상 유도 플랫폼의 (B)의 측면보기. (C) mTBI의 유도에 사용되는 무게의 사진 데모 . 업로드 / 51820 / 51820fig3highres.jpg &quot;폭 =&quot;500 &quot;/&gt; 그림 3 :. mTBI 전시에게 상당한 증가를받은 단일 mTBI와 가짜 부상을 경험 한 청소년 쥐 (* P &lt;0.01) 쥐를 경험 한 시간에서 오른쪽으로 청소년 쥐 사이의 평균 차이의 그래픽 표시 시간의 지속 시간은 앙와위에서 스스로를 잘 할 필요가 있었다. <img alt="그림 4" fo:content-width="5in" src="/files/ftp_upload/51820/51820fig4highres.jpg" width="500" /> 그림 4 : 단일 mTBI와 가짜 부상을 경험 한 청소년 쥐 (* P &lt;0.05)을 경험 한 청소년 쥐에 의해 빔 산책 작업에 전시 뒷다리 발 - 전표의 평균 개수의 그래픽 표현입니다.

Watch this Scientific Journal Video about A Novel Model of Mild Traumatic Brain Injury for Juvenile Rats at JoVE.com

A Novel Model of Mild Traumatic Brain Injury for Juvenile Rats

The modified weight-drop technique is an easy, cost-effective procedure used for the induction of mild traumatic brain injury in juvenile rats. This novel technique produces clinically relevant symptomology that will advance the study of mild traumatic brain injury (mTBI) and concussion.

청소년 쥐에 대한 가벼운 외상성 뇌 손상의 소설 모델

Despite growing evidence that childhood represents a major risk period for mild traumatic brain injury (mTBI) from sports-related concussions, motor ...

a-novel-model-of-mild-traumatic-brain-injury-for-juvenile-rats

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JoVE Journal

Medicine

23.3K 조회수.  University of Calgary. The modified weight-drop technique is an easy, cost-effective procedure used for the induction of mild traumatic brain injury in juvenile rats. This novel technique produces clinically relevant symptomology that will advance the study of mild traumatic brain injury (mTBI) and concussion. 

비디오: A Novel Model of Mild Traumatic Brain Injury for Juvenile Rats

Acute and Chronic Tactile Sensory Testing after Spinal Cord Injury in Rats

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory testing in rat SCI models is needed. However, until recently, no single testing approach had been validated for SCI so that standardized methods have not been implemented across labs. Additionally, available testing methods could not be implemented acutely or when severe motor impairments existed, preventing studies of the development of SCI-induced allodynia3. Here we present two validated sensory testing methods using von Frey Hair (VFH) monofilaments which quantify changes in tactile sensory thresholds after SCI4-5. One test is the well-established Up-Down test which demonstrates high sensitivity and specificity across different SCI severities when tested chronically5. The other test is a newly-developed dorsal VFH test that can be applied acutely after SCI when allodynia develops, prior to motor recovery4-5. Each VFH monofilament applies a calibrated force when touched to the skin of the hind paw until it bends. In the up-down method, alternating VFHs of higher or lower forces are used on the plantar L5 dermatome to delineate flexor withdrawal thresholds. Successively higher forces are applied until withdrawal occurs then lower force VFHs are used until withdrawal ceases. The tactile threshold reflects the force required to elicit withdrawal in 50% of the stimuli. For the new test, each VFH is applied to the dorsal L5 dermatome of the paw while the rat is supported by the examiner. The VFH stimulation occurs in ascending order of force until at least 2 of 3 applications at a given force produces paw withdrawal. Tactile sensory threshold is the lowest force to elicit withdrawal 66% of the time. Acclimation, testing and scoring procedures are described. Aberrant trials that require a retest and typical trials are defined. Animal use was approved by Ohio State University Animal Care and Use Committee.

We describe two tactile sensory testing methods for acute or chronic periods of spinal cord injury in rats. These validated procedures can detect the development and maintenance of allodynia-like sensations.

쥐의 척수를 다친 후에 급성 및 만성 촉각 감각 테스트

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory ...

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A Contusion Model of Severe Spinal Cord Injury in Rats

The translational potential of novel treatments should be investigated in severe spinal cord injury (SCI) contusion models. A detailed methodology is described to obtain a consistent model of severe SCI. Use of a stereotactic frame and computer controlled impactor allows for creation of reproducible injury. Hypothermia and urinary tract infection pose significant challenges in the post-operative period. Careful monitoring of animals with daily weight recording and bladder expression allows for early detection of post-operative complications. The functional results of this contusion model are equivalent to transection models. The contusion model can be utilized to evaluate the efficacy of both neuroprotective and neuroregenerative approaches.

A contusion model of severe spinal cord injury is described. Detailed pre-operative, operative and post-operative steps are described to obtain a consistent model.

쥐에 심각한 척수 손상의 타박상 모델

The translational potential of novel treatments should be investigated in severe spinal cord injury (SCI) contusion models. A detailed methodology is ...

A Novel Vertebral Stabilization Method for Producing  Contusive Spinal Cord Injury

Clinically-relevant animal cervical spinal cord injury (SCI) models are essential for developing and testing potential therapies; however, producing reliable cervical SCI is difficult due to lack of satisfactory methods of vertebral stabilization. The conventional method to stabilize the spine is to suspend the rostral and caudal cervical spine via clamps attached to cervical spinous processes. &#160;However, this method of stabilization fails to prevent tissue yielding during the contusion as the cervical spinal processes are too short to be effectively secured by the clamps (Figure 1). &#160;Here we introduce a new method to completely stabilize the cervical vertebra at the same level of the impact injury. &#160;This method effectively minimizes movement of the spinal column at the site of impact, which greatly improves the production of consistent SCIs. &#160;We provide visual description of the equipment (Figure 2-4), methods, and a step-by-step protocol for the stabilization of the cervical 5 vertebra (C5) of adult rats, to perform laminectomy (Figure 5) and produce a contusive SCI thereafter. &#160;Although we only demonstrate a cervical hemi-contusion using the NYU/MASCIS impactor device, this vertebral stabilization technique can be applied to other regions of the spinal cord, or be adapted to other SCI devices. &#160;Improving spinal cord exposure and fixation through vertebral stabilization may be valuable for producing consistent and reliable injuries to the spinal cord. &#160;This vertebral stabilization method can also be used for stereotactic injections of cells and tracers, and for imaging using two-photon microscopy in various neurobiological studies.

Vertebral stabilization is necessary for minimizing variability, and for producing consistent experimental spinal cord injuries.&#160; Using a customized stabilizing apparatus in conjunction with the NYU/MASCIS impactor device, we have demonstrated here the proper equipment and procedure for generating reproducible hemi-contusive cervical (C5) spinal cord injuries in adult rats.

Contusive 척수 손상의 제조 소설 척추 안정화 방법

Clinically-relevant animal cervical spinal cord injury (SCI) models are essential for developing and testing potential therapies; however, producing ...

Stretch in Brain Microvascular Endothelial Cells (cEND) as an In Vitro Traumatic Brain Injury Model of the Blood Brain Barrier

Due to the high mortality incident brought about by traumatic brain injury (TBI), methods that would enable one to better understand the underlying mechanisms involved in it are useful for treatment. There are both in vivo and in vitro methods available for this purpose. In vivo models can mimic actual head injury as it occurs during TBI. However, in vivo techniques may not be exploited for studies at the cell physiology level. Hence, in vitro methods are more advantageous for this purpose since they provide easier access to the cells and the extracellular environment for manipulation.
Our protocol presents an in vitro model of TBI using stretch injury in brain microvascular endothelial cells. It utilizes pressure applied to the cells cultured in flexible-bottomed wells. The pressure applied may easily be controlled and can produce injury that ranges from low to severe. The murine brain microvascular endothelial cells (cEND) generated in our laboratory is a well-suited model for the blood brain barrier (BBB) thus providing an advantage to other systems that employ a similar technique. In addition, due to the simplicity of the method, experimental set-ups are easily duplicated. Thus, this model can be used in studying the cellular and molecular mechanisms involved in TBI at the BBB.

Stretch in Brain Microvascular Endothelial Cells cEND as an In Vitro Traumatic Brain Injury Model of the Blood Brain Barrier

In vitro traumatic brain injury models are being developed to reproduce in vivo brain deformation. Stretch-induced injury has been employed for astrocytes, neurons, glial cells, aortic, and brain endothelial cells. However, our system uses a blood brain barrier (BBB) model that possesses properties constituting a legitimate model of the BBB to establish an in vitro&#160;TBI model.

로 뇌 미세 혈관 내피 세포 (CEnd를)에 기지개<em&gt; 체외</em혈액 뇌 장벽의&gt; 외상성 뇌 손상 모델

Due to the high mortality incident brought about by traumatic brain injury (TBI), methods that would enable one to better understand the underlying ...

Controlled Cortical Impact Model for Traumatic Brain Injury

Every year over a million Americans suffer a traumatic brain injury (TBI). Combined with the incidence of TBIs worldwide, the physical, emotional, social, and economical effects are staggering. Therefore, further research into the effects of TBI and effective treatments is necessary. The controlled cortical impact (CCI) model induces traumatic brain injuries ranging from mild to severe. This method uses a rigid impactor to deliver mechanical energy to an intact dura exposed following a craniectomy. Impact is made under precise parameters at a set velocity to achieve a pre-determined deformation depth. Although other TBI models, such as weight drop and fluid percussion, exist, CCI is more accurate, easier to control, and most importantly, produces traumatic brain injuries similar to those seen in humans. However, no TBI model is currently able to reproduce pathological changes identical to those seen in human patients. The CCI model allows investigation into the short-term and long-term effects of TBI, such as neuronal death, memory deficits, and cerebral edema, as well as potential therapeutic treatments for TBI.

Traumatic brain injuries (TBIs) remain a serious health problem. Using the controlled cortical impact surgery model, research on the effects of TBI and possible treatment methods may be performed.

외상성 뇌 손상에 대한 제어 피질 충격 모델

Every year over a million Americans suffer a traumatic brain injury (TBI).  Combined with the incidence of TBIs worldwide, the physical, emotional, ...

Renal Ischaemia Reperfusion Injury: A Mouse Model of Injury and Regeneration

Renal ischaemia reperfusion injury (IRI) is a common cause of acute kidney injury (AKI) in patients and occlusion of renal blood flow is unavoidable during renal transplantation. Experimental models that accurately and reproducibly recapitulate renal IRI are crucial in dissecting the pathophysiology of AKI and the development of novel therapeutic agents. Presented here is a mouse model of renal IRI that results in reproducible AKI. This is achieved by a midline laparotomy approach for the surgery with one incision allowing both a right nephrectomy that provides control tissue and clamping of the left renal pedicle to induce ischaemia of the left kidney. By careful monitoring of the clamp position and body temperature during the period of ischaemia this model achieves reproducible functional and structural injury. Mice sacrificed 24 hr&#160;following surgery demonstrate loss of renal function with elevation of the serum or plasma creatinine level as well as structural kidney damage with acute tubular necrosis evident. Renal function improves and the acute tissue injury resolves during the course of 7 days following renal IRI such that this model may be used to study renal regeneration. This model of renal IRI has been utilized to study the molecular and cellular pathophysiology of AKI as well as analysis of the subsequent renal regeneration.

The mouse model of renal ischaemia reperfusion injury described here comprises of a right nephrectomy that provides control tissue and clamping of the left renal pedicle to induce ischaemia that results in acute kidney injury. This model uses a midline laparotomy approach with all steps performed via one incision.

신장 허혈 재관류 상해 : 상해의 마우스 모델 및 재생

Renal ischaemia reperfusion injury (IRI) is a common cause of acute kidney injury (AKI) in patients and occlusion of renal blood flow is unavoidable ...

A Tissue Displacement-based Contusive Spinal Cord Injury Model in Mice

Producing a consistent and reproducible contusive spinal cord injury (SCI) is critical to minimizing behavioral and histological variabilities between experimental animals. Several contusive SCI models have been developed to produce injuries using different mechanisms. The severity of the SCI is based on the height that a given weight is dropped, the injury force, or the spinal cord displacement. In the current study, we introduce a novel mouse contusive SCI device, the Louisville Injury System Apparatus (LISA) impactor, which can create a displacement-based SCI with high injury velocity and accuracy. This system utilizes laser distance sensors combined with advanced software to produce graded and highly-reproducible injuries. We performed a contusive SCI at the 10th thoracic vertebral (T10) level in mice to demonstrate the step-by-step procedure. The model can also be applied to the cervical and lumbar spinal levels.

We introduce a tissue displacement-based contusive spinal cord injury model that can produce a consistent contusive spinal cord injury in adult mice.

마우스의 조직 변위에 근거한 척수 손상 모델

Producing a consistent and reproducible contusive spinal cord injury (SCI) is critical to minimizing behavioral and histological variabilities between ...

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 ...

Spinal Cord Lateral Hemisection and Asymmetric Behavioral Assessments in Adult Rats

Incomplete spinal cord injury (SCI) often leads to impairments of sensorimotor functions and is clinically the most frequent type of SCI. Human Brown-S&#233;quard syndrome&#160;is a common type of incomplete SCI caused by a lesion to one half of the spinal cord which results in paralysis and loss of&#160;proprioception&#160;on the same (or ipsilesional) side as the injury, and loss of pain and temperature sensation on the opposite (or contralesional) side. Adequate methodologies for producing a spinal cord lateral hemisection (HX) and assessing neurological impairments are essential to establish a reliable animal model of Brown-S&#233;quard syndrome. Although lateral hemisection model plays a pivotal role in basic and translational research, standardized protocols for creating such a hemisection and assessing unilateralized function are lacking. The goal of this study is to describe step-by-step procedures to produce a rat spinal lateral HX at the 9th thoracic (T9) vertebral level. We, then, describe a combined behavior scale for HX (CBS-HX) that provides a simple and sensitive assessment of asymmetric neurological performance for unilateral SCI. The CBS-HX, ranging from 0 to 18, is composed of 4 individual assessments which include unilateral hindlimb stepping (UHS), coupling, contact placing, and grid walking. For CBS-HX, the ipsilateral and contralateral hindlimbs are assessed separately. We found that, after a T9 HX, the ipsilateral hindlimb showed impaired behavior function whereas the contralateral hindlimb showed substantial recovery. The CBS-HX effectively discriminated behavioral functions between ipsilateral and contralateral hindlimbs and detected temporal progression of recovery of the ipsilateral hindlimb. The CBS-HX components can be analyzed separately or in combination with other measures when needed. Although we only provided visual descriptions of the surgical procedures and behavioral assessments of a thoracic HX, the principle may be applied to other incomplete SCIs and at other levels of the injury.

Here we describe surgical procedures to produce a reliable spinal cord lateral hemisection (HX) at the 9th thoracic level in adult rats and neurobehavioral assessments designed for detecting asymmetric deficits after such a unilateral injury.

성인 쥐의 척수 측삭 절제술 및 비대칭 행동 평가

Incomplete spinal cord injury (SCI) often leads to impairments of sensorimotor functions and is clinically the most frequent type of SCI. Human ...

Semi-quantitative Assessment Using [18F]FDG Tracer in Patients with Severe Brain Injury

Patients with severe traumatic brain injury (sTBI) have difficulty knowing whether they are accurately expressing their thoughts and emotions because of disorders of consciousness, disrupted higher brain function, and verbal disturbances. As a consequence of an insufficient ability to communicate, objective evaluations are needed from family members, medical staff, and caregivers. One such evaluation is the assessment of functioning brain areas. Recently, multimodal brain imaging has been used to explore the function of damaged brain areas. [18F]-fluorodeoxyglucose positron emission tomography-computed tomography ([18F]FDG-PET/CT) is a successful tool for examining brain function. However, the assessment of brain glucose metabolism based on [18F]FDG-PET/CT is not standardized and depends on several varying parameters, as well as the patient&#39;s condition. Here, we describe a series of semiquantitative assessment protocols for a region-of-interest (ROI) image analysis using self-produced [18F]FDG tracers in patients with sTBI. The protocol focuses on screening the participants, preparing the [18F]FDG tracer in the hot lab, scheduling the acquisition of [18F]FDG-PET/CT brain images, and measuring glucose metabolism using the ROI analysis from a targeted brain area.

Semi-quantitative Assessment Using [18F]FDG Tracer in Patients with Severe Brain Injury

[18F]-fluorodeoxyglucose (FDG) positron emission tomography-computed tomography is useful for studying glucose metabolism related to brain function. Here, we present a protocol for an [18F]FDG tracer set-up and semiquantitative assessment of the region-of-interest analysis for targeted brain areas associated with clinical manifestations in patients with severe traumatic brain injury.

심한 뇌 손상 환자에서 [18F] FDG 추적 프로그램을 사용 하 여 반 정량적 평가

Patients with severe traumatic brain injury (sTBI) have difficulty knowing whether they are accurately expressing their thoughts and emotions because of ...