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>

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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.

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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 ...

a-repetitive-concussive-head-injury-model-in-mice

Research

JoVE Journal

Medicine

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

The Spared Nerve Injury (SNI) Model of Induced Mechanical Allodynia in Mice

Peripheral neuropathic pain is a severe chronic pain condition which may result from trauma to sensory nerves in the peripheral nervous system. The spared nerve injury (SNI) model induces symptoms of neuropathic pain such as mechanical allodynia i.e. pain due to tactile stimuli that do not normally provoke a painful response [1]. 
The SNI mouse model involves ligation of two of the three branches of the sciatic nerve (the tibial nerve and the common peroneal nerve), while the sural nerve is left intact [2]. The lesion results in marked hypersensitivity in the lateral area of the paw, which is innervated by the spared sural nerve. The non-operated side of the mouse can be used as a control. The advantages of the SNI model are the robustness of the response and that it doesn&#x2019;t require expert microsurgical skills.
The threshold for mechanical pain response is determined by testing with von Frey filaments of increasing bending force, which are repetitively pressed against the lateral area of the paw [3], [4]. A positive pain reaction is defined as sudden paw withdrawal, flinching and/or paw licking induced by the filament. A positive response in three out of five repetitive stimuli is defined as the pain threshold. 
As demonstrated in the video protocol, C57BL/6 mice experience profound allodynia as early as the day following surgery and maintain this for several weeks.

The Spared Nerve Injury SNI Model of Induced Mechanical Allodynia in Mice

The Spared Nerve Injury animal model is described here as a mouse model of peripheral neuropathic pain following partial denervation of the sciatic nerve by lesioning the tibial and common peroneal nerve branches, leaving the remaining sural nerve intact. Behavioral modification resulting from mechanical allodynia is quantified by von Frey filaments.

마우스에서 유도된 기계 Allodynia의 겪었던 신경 상해 (스니) 모델

Peripheral neuropathic pain is a severe chronic pain condition which may result from trauma to sensory nerves in the peripheral nervous system. The spared ...

연구

의학

Controlled Cervical Laceration Injury in Mice

Use of genetically modified mice enhances our understanding of molecular mechanisms underlying several neurological disorders such as a spinal cord injury (SCI). Freehand manual control used to produce a laceration model of SCI creates inconsistent injuries often associated with a crush or contusion component and, therefore, a novel technique was developed. Our model of cervical laceration SCI has resolved inherent difficulties with the freehand method by incorporating 1) cervical vertebral stabilization by vertebral facet fixation, 2) enhanced spinal cord exposure, and 3) creation of a reproducible laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm in depth without associated contusion. Compared to the standard methods of creating a SCI laceration such as freehand use of a scalpel or scissors, our method has produced a consistent lesion. This method is useful for studies on axonal regeneration of corticospinal, rubrospinal, and dorsal ascending tracts.

A novel technique to create a reproducible in vivo model of cervical spinal cord laceration injury in the mouse is described. This technique is based on spine stabilization by fixation of the cervical facets and laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm.

마우스의 제어 자궁 경부 열상 상해

Use of genetically modified mice enhances our  understanding of molecular mechanisms underlying several neurological disorders  such as a spinal cord ...

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 ...

Ischemia-reperfusion Model of Acute Kidney Injury and Post Injury Fibrosis in Mice

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often unreported mortality rates that may confound analyses. Bilateral renal pedicle clamping is commonly used to induce IR-AKI, but differences between effective clamp pressures and/or renal responses to ischemia between kidneys often lead to more variable results. In addition, shorter clamp times are known to induce more variable tubular injury, and while mice undergoing bilateral injury with longer clamp times develop more consistent tubular injury, they often die within the first 3 days after injury due to severe renal insufficiency. To improve post-injury survival and obtain more consistent and predictable results, we have developed two models of unilateral ischemia-reperfusion injury followed by contralateral nephrectomy. Both surgeries are performed using a dorsal approach, reducing surgical stress resulting from ventral laparotomy, commonly used for mouse IR-AKI surgeries. For induction of moderate injury BALB/c mice undergo unilateral clamping of the renal pedicle for 26 min and also undergo simultaneous contralateral nephrectomy. Using this approach, 50-60% of mice develop moderate AKI 24 hr after injury but 90-100% of mice survive. To induce more severe AKI, BALB/c mice undergo renal pedicle clamping for 30 min followed by contralateral nephrectomy 8 days after injury. This allows functional assessment of renal recovery after injury with 90-100% survival. Early post-injury tubular damage as well as post injury fibrosis are highly consistent using this model.

We describe models of moderate and severe ischemia-reperfusion-induced kidney injury in which the mice undergo unilateral renal pedicle clamping followed by simultaneous or delayed contralateral nephrectomy, respectively. These models consistently give rise to renal dysfunction and post-injury fibrosis, but injury severity and survival are dependent on mouse background, age and surgical equipment.

마우스의 급성 신장 손상 및 사후 상해 섬유증의 허혈 - 재관류 모델

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often ...

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 Simple Critical-sized Femoral Defect Model in Mice

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all limb bone fractures fail to heal completely due to the extent of the trauma, disease, or age of the patient. Our ability to improve bone regenerative strategies is critically dependent on the ability to mimic serious bone trauma in test animals, but the generation and stabilization of large bone lesions is technically challenging. In most cases, serious long bone trauma is mimicked experimentally by establishing a defect that will not naturally heal. This is achieved by complete removal of a bone segment that is larger than 1.5 times the diameter of the bone cross-section. The bone is then stabilized with a metal implant to maintain proper orientation of the fracture edges and allow for mobility. 
Due to their small size and the fragility of their long bones, establishment of such lesions in mice are beyond the capabilities of most research groups. As such, long bone defect models are confined to rats and larger animals. Nevertheless, mice afford significant research advantages in that they can be genetically modified and bred as immune-compromised strains that do not reject human cells and tissue.
Herein, we demonstrate a technique that facilitates the generation of a segmental defect in mouse femora using standard laboratory and veterinary equipment. With practice, fabrication of the fixation device and surgical implantation is feasible for the majority of trained veterinarians and animal research personnel. Using example data, we also provide methodologies for the quantitative analysis of bone healing for the model.

Animal models are frequently employed to mimic serious bone injury in biomedical research. Due to their small size, establishment of stabilized bone lesions in mice are beyond the capabilities of most research groups. Herein, we describe a simple method for establishing and analyzing experimental femoral defects in mice.

마우스의 간단한 임계 크기의 대퇴 결함 모델

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all ...

Facial Nerve Axotomy in Mice: A Model to Study Motoneuron Response to Injury

The goal of this surgical protocol is to expose the facial nerve, which innervates the facial musculature, at its exit from the stylomastoid foramen and either cut or crush it to induce peripheral nerve injury. Advantages of this surgery are its simplicity, high reproducibility, and the lack of effect on vital functions or mobility from the subsequent facial paralysis, thus resulting in a relatively mild surgical outcome compared to other nerve injury models. A major advantage of using a cranial nerve injury model is that the motoneurons reside in a relatively homogenous population in the facial motor nucleus in the pons, simplifying the study of the motoneuron cell bodies. Because of the symmetrical nature of facial nerve innervation and the lack of crosstalk between the facial motor nuclei, the operation can be performed unilaterally with the unaxotomized side serving as a paired internal control. A variety of analyses can be performed postoperatively to assess the physiologic response, details of which are beyond the scope of this article. For example, recovery of muscle function can serve as a behavioral marker for reinnervation, or the motoneurons can be quantified to measure cell survival. Additionally, the motoneurons can be accurately captured using laser microdissection for molecular analysis. Because the facial nerve axotomy is minimally invasive and well tolerated, it can be utilized on a wide variety of genetically modified mice. Also, this surgery model can be used to analyze the effectiveness of peripheral nerve injury treatments. Facial nerve injury provides a means for investigating not only motoneurons, but also the responses of the central and peripheral glial microenvironment, immune system, and target musculature. The facial nerve injury model is a widely accepted peripheral nerve injury model that serves as a powerful tool for studying nerve injury and regeneration.

We present a surgical protocol detailing how to perform a cut or crush axotomy on the facial nerve in the mouse. The facial nerve axotomy can be employed to study the physiological response to nerve injury and test therapeutic techniques.

마우스의 얼굴 신경 Axotomy : 부상으로 운동 신경 응답을 연구하는 모델

The goal of this surgical protocol is to expose the facial nerve, which innervates the facial musculature, at its exit from the stylomastoid foramen and ...

Quantification of Neurovascular Protection Following Repetitive Hypoxic Preconditioning and Transient Middle Cerebral Artery Occlusion in Mice

Experimental animal models of stroke are invaluable tools for understanding stroke pathology and developing more effective treatment strategies. A 2 week protocol for repetitive hypoxic preconditioning (RHP) induces long-term protection against central nervous system (CNS) injury in a mouse model of focal ischemic stroke. RHP consists of 9 stochastic exposures to hypoxia that vary in both duration (2 or 4 hr) and intensity (8% and 11% O2). RHP reduces infarct volumes, blood-brain barrier (BBB) disruption, and the post-stroke inflammatory response for weeks following the last exposure to hypoxia, suggesting a long-term induction of an endogenous CNS-protective phenotype. The methodology for the dual quantification of infarct volume and BBB disruption is effective in assessing neurovascular protection in mice with RHP or other putative neuroprotectants. Adult male Swiss Webster mice were preconditioned by RHP or duration-equivalent exposures to 21% O2 (i.e. room air). A 60 min transient middle cerebral artery occlusion (tMCAo) was induced 2 weeks following the last hypoxic exposure. Both the occlusion and reperfusion were confirmed by transcranial laser Doppler flowmetry. Twenty-two hr after reperfusion, Evans Blue (EB) was intravenously administered through a tail vein injection. 2 hr later, animals were sacrificed by isoflurane overdose and brain sections were stained with 2,3,5- triphenyltetrazolium chloride (TTC). Infarcts volumes were then quantified. Next, EB was extracted from the tissue over 48 hr to determine BBB disruption after tMCAo. In summary, RHP is a simple protocol that can be replicated, with minimal cost, to induce long-term endogenous neurovascular protection from stroke injury in mice, with the translational potential for other CNS-based and systemic pro-inflammatory disease states.

This protocol describes repetitive hypoxic preconditioning, or brief exposures to systemic hypoxia that reduce infarct volumes and blood-brain barrier disruption following transient middle cerebral artery occlusion in mice. It also details dual quantification of infarct volume and blood-brain barrier disruption after stroke to assess the efficacy of neurovascular protection.

마우스의 반복 저산소 전처리 및 과도 중간 대뇌 동맥 폐쇄 후 신경 혈관 보호의 정량화

Experimental animal models of stroke are invaluable tools for understanding stroke pathology and developing more effective treatment strategies. A 2 week ...

Photothrombosis-induced Focal Ischemia as a Model of Spinal Cord Injury in Mice

Spinal cord injury (SCI) is a devastating clinical condition causing permanent changes in sensorimotor and autonomic functions of the spinal cord (SC) below the site of injury. The secondary ischemia that develops following the initial mechanical insult is a serious complication of the SCI and severely impairs the function and viability of surviving neuronal and non-neuronal cells in the SC. In addition, ischemia is also responsible for the growth of lesion during chronic phase of injury and interferes with the cellular repair and healing processes. Thus there is a need to develop a spinal cord ischemia model for studying the mechanisms of ischemia-induced pathology. Focal ischemia induced by photothrombosis (PT) is a minimally invasive and very well established procedure used to investigate the pathology of ischemia-induced cell death in the brain. Here, we describe the use of PT to induce an ischemic lesion in the spinal cord of mice. Following retro-orbital sinus injection of Rose Bengal, the posterior spinal vein and other capillaries on the dorsal surface of SC were irradiated with a green light resulting in the formation of a thrombus and thus ischemia in the affected region. Results from histology and immunochemistry studies show that PT-induced ischemia caused spinal cord infarction, loss of neurons and reactive gliosis. Using this technique a highly reproducible and relatively easy model of SCI in mice can be achieved that would serve the purpose of scientific investigations into the mechanisms of ischemia induced cell death as well as the efficacy of neuroprotective drugs. This model will also allow exploration of the pathological changes that occur following SCI in live mice like axonal degeneration and regeneration, neuronal and astrocytic Ca2+ signaling using two-photon microscopy.

Photothrombosis is a minimally invasive and highly reproducible procedure to induce focal ischemia in the spinal cord and serves as a model of spinal cord injury in mice.

쥐의 척수 손상의 모델로 초점 국소 빈혈을 Photothrombosis는 유도

Spinal cord injury (SCI) is a devastating clinical condition causing permanent changes in sensorimotor and autonomic functions of the spinal cord (SC) ...

Induction of Accelerated Atherosclerosis in Mice: The "Wire-Injury" Model

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. Moreover, by rupture of the defective vascular wall, atherosclerosis induces occlusive thrombus formation, which represents the main cause of myocardial infarction or stroke and the most frequent cause of death. Despite the advances in the cardiovascular field, many questions remain unanswered, and additional basic research is essential to improve our understanding of the molecular mechanisms during atherosclerosis and its effects. Due to limited clinical studies, there is a need for representative animal models recreating atherosclerotic conditions such as neointima formation after stent implantation, balloon angioplasty, or endarterectomy. Since the mouse presents many advantages and is the most frequently used model for studying molecular processes, the current study proposes an invasive procedure of endothelial denudation, also known as the wire-injury model, which is representative of the human condition of neointima formation in arteries after revascularization procedures.

This study describes an invasive procedure for the induction of accelerated atherosclerosis in mice. In comparison to other methods using electric- or cryo-induced injury, mechanical-induced injury mimics the human condition of restenosis after revascularization therapies and is ideal for the study of the molecular mechanisms involved.

마우스에서 가속 죽상 경화증의 유도: "와이어 부상" 모델

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. ...