저자는 박사 N. Banik이 모델의 개발에 자신의 지침 박사 D. 미첼에게 감사합니다.

1. 척수 손상 전 준비 이 절차에 필요한 수술 도구, 치아, hemostats, 자체 유지 견인기, 좋은 팁 rongeurs, 바늘 드라이버, 흡수성 봉합, 피부 클립 도포와와없는 픽업 메스입니다. 필요한 다른 수술 용품 수술 커튼, 수술 필드 멸균 시트, 거즈 스폰지, 면봉 도포 및 금속 호일입니다. 수술하기 전에 수술 장비 및 소모품을 압력솥. 하나의 동물에 대한 장비 및 소모품의 개별 집합을 사용합니다. 외과 영역 및 알코올 잎사귀와 장치 (충격기, 광원, 정위 프레임, 유리 구슬 살균 및 가열 패드) 청소합니다. 수술 커튼을 열고 조심스럽게 오염을 방지 수술 필드를 드레이프 멸균 장갑을 사용한다. 무균 개별 악기를 열고 조심스럽게 수술 분야의 사람들을 배치합니다. 일의 손잡이와 핸들 커버전자 장치는 가능성 멸균 금속 호일 절차를 수행하는 동안 필요합니다. 스위치에 유리 구슬 살균기 절차를 수행하는 동안 사용하기 위해 준비합니다. 2. 동물의 준비 실제 절차 전에 실험실 영역 몇 시간에 쥐를 가져옵니다. 앞으로 기대 절차 (일반적으로 0.006 밀리그램 / ML 피하의 buprinorphine 1.5 ml)에 적어도 하나의 시간 진통제를 관리 할 수​​ 있습니다. 수술 전 항생제 (일반적으로 Baytril 4 ㎎ / kg 피하) 관리 할 수​​ 있습니다. 90 ㎎ / 케타민의 kg을 사용하여 쥐를 마취하고 아무 발가락 핀치 응답이 없을 때까지 기다립니다. 흉추에서 가장 눈에 띄는 극돌기를 만져. 이 수준은 일반적으로 T10 극돌기에 해당합니다. T10과 관련하여 구성 수준의 위치를​​ 표시합니다. 우리가 실험실에서 우리는 T10 상해를 수행합니다. 다음 이야기는 T10 SCI의 기술을 설명합니다. 길이 방향 3cm x 6 센티미터 사각형을 면도 T10 수준을 중심으로. 피부를 세 번 위스콘신 청소번째 베타 딘 용액. 각 눈에 안과 윤활제를 적용합니다. 주의 깊게 오염을 방지 수술 필드에 편안한 위치에 쥐를 전송합니다. 코어 온도를 모니터링하고 주변 정상 (~ 37.5 ° C) 가능한에 관해서는 동물의 온도를 유지하기 위해 적절하게 난방을 조정하는 직장​​ 온도 프로브를 삽입합니다. 수술 부위 위의 창 수술 장식 주름 쥐를 다룹니다. 3. 수술 T10 마크를 중심으로 # 10 블레이드를 사용하여 대략 4cm 절개로 시작합니다. 참을성 멀리 T9-T11 극돌기 프로세스와 엽층의 근막과 근육 레이어를 해부를 진행합니다. 뼈 근육과 근막을 멀리 철회 견인기를 놓습니다. T9의 극돌기를 안정화하는 동안 크게 고급 가위를 사용하여 T9와 T10 사이의 극간 인대를 나눕니다. 마찬가지로 T10 극돌기를 안정시키고 T10-T11 사이의 극간 인대를 나눕니다. 보완하기 위해 루프 배율을 사용인대 flavum (그림 1)를 통해 테 인대의 부분은 모든 방법. 인대 flavum가 연결되면 thecal 낭은 명백하다. 주의 T10에서 양측 조각 - 식사 후궁을 수행하기 위해 좋은 팁 rongeurs를 사용합니다. 정성이 rongeur 끝에서 thecal 주머니 및 부주의 부상에 하향 압력을 방지하기 위해 수행됩니다. 라미와 T10의 극돌기가 완전히 제거됩니다. 안정화 플랫폼의 위치로 동물을 이동합니다. T9 척추의 측면 측면 뒤에 T11 척추의 측면 측면을 체결하여 척추를 고정시키기 위해 안정 클램프를 사용합니다. 이 호흡 운동을위한 공간을 제한하고 원치 않는 스트레스에게 동물을 추가로 안정화 클램프 플랫폼에 쥐를 압축하지 않도록주의하십시오. 확보 한 후 컴퓨터 제어 충격기에 척추 설정이 확인됩니다. 우리는 일반적으로 SP에 3.0 mm의 임팩터 팁을 사용하여2mm의 깊이와 0.3 초​​ 드웰 시간에 4cm / S의 EED. 임팩터 팁을 확장하고 그냥 코드 표면에 닿을 때까지 내립니다. 끝을 철회하고 척수 표면을 향해 장치 2mm를 내립니다. 심한 타박상 척수 손상을 일으킬 4cm / s의 속도로 피스톤을 해제합니다. 코드에 불필요한 압력을 생성하지 않도록주의 대신에면 끝 도포를 유지하기 위해 충분한 압력을 사용하여 지혈을 달성. 그림 - 8 스티치가 조심로 봉합 근육과 근막 층은 흡수성 봉합사를 사용하여 너무 꽉 당겨하지. 2 개의 작은 스테이플의 최소 피부를 닫습니다, 절개 부분은 첫 번째 2 또는 3 스테이플 후 열려있는 경우 최대 4 ~ 5 스테이플을 사용할 수있다. 3. 시술 후 관리 24 시간 후 수술에 대해 33 ~ 35 ° C의 따뜻한 환경에서 장소 쥐. 그들이 움직이기 시작되면이 인큐베이터 (그들은 의식이있는 동안) 및 가열 케이지 공간을 수반한다. 일단 쥐가 완전히 아입니다아케는 식염수의 피하 buprinorphine의 1.5 ML (0.006 밀리그램 / ML) 및 Baytril 0.1 mL를 5ml를 관리. 칠일 동안 하루에 한 번 첫 24-48 시간 및 Baytril 위해 피하 하루에 두 번 buprinorphine를 계속합니다. 방광은 수동으로 방광 기능의 반환 (3 일 연속 이른 아침 식 소변 &lt;2 ㎖)까지 하루에 세 번 표현해야한다. 동물도 감염이 시간 (소변에 혈액, 흰 색, 악취) 중에 선택해야 신체 활동 또는 상처 치유 문제를 감소. 감염의 존재는 지역 수의사와 상담 항생제의 증가 용량 (또는 다시 시작)를 반대해야한다. 매일 자신의 회복을 평가하기 위해 수술 다음날부터 쥐 무게.

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재정 공시 없음 자금 유출 없음.

여러 가지 새로운 치료는 최근 SCI 연구 분야의 초기 약속을 보여 주었다. 이러한 치료 3주의 평가는 최대 번역 가능성 전략을 선택하는 SCI의 임상 적 모델에 필수적입니다. 등급의 계획은 최근 전임상 연구의 강도를 평가하기 위해 개발되었다. 9이 제도는 심각한 SCI의 타박상 모델을 활용의 중요성을 강조했다. 여기에 우리가 절개 모델들을 닮은 일관된 병변 볼륨 및 기능 점수가 ?...

적절한 손상 모델의 선택은 척수 손상에 대한 새로운 치료법 (SCI)의 전임상 평가에 매우 중요합니다. 의사와 neurotrauma 타박상 모델 분야의 과학자의 최근 조사에 1,2,13, hemisection 또는 전체 절개 모델에 반대 보편적으로 임상 적으로 받아 들여졌다. 8이 견해는 인간의 척수 손상의 대부분은 자연 contusive 것을 관찰을 기반으로합니다. 10 타박상의 생물학은 hemisection 또는 절개 모델에서 다른 것으로 보인다. 11 Iseda, 등이 알이. hemisection 및 타박상 모델에서 별도 neuroregeneration에 intraspinal chondroitinase ABC 주입의 효과를 비교 하였다. 4 축삭 재생이 hemisection에 신경 다리에서 관찰하지만, 타박상 SCI 그룹되지 않았습니다. hemisection 또는 전체 절개 모델은 또한 CL의 단지 아주 작은 하위 집합에 존재하는 것으로 알려진 조건을 만들inical 상황. 예를 들어, 몇몇 연구자들은 hemisection 또는 재생을 촉진하는 완전 절개 후 병변 캐비티에 주입을위한 발판 기반의 개입을 고용했다. 6 손상된 척수 내에서 공동의 생성은 비실용적이고 아마 비 윤리적이기 때문에이 방법은 임상 적으로 관련이됩니다. 기능 회복의 변화는 타박상 모델의 주요 과제 남아있다. 5,12이 변화는 척수 볼륨에 걸쳐 균일 한 힘의 전달에 영향을 미치는 특히 ventrally있는 모터 경로 전에 컴퓨터 제어 충격기와 척추 안정화의 사용으로 최소화 할 수있다 . 그것은 생존 축삭에서 소성 및 담보 비중이 척수 손상 후 회복의 주된 메커니즘 주목해야합니다. 타박상 기술 1 따라서도 약간의 변동이 크게 다른 결과를 얻을 수 있습니다. 이를 위해 우리는 개발일관성 타박상 볼륨과 절개 모델과 비교 기능 회복을 산출 심각한 척수 손상 모델. 이 모델은 치료 효과에 대한 개념의 증거로 신경 보호 및 neuroregeneration 전략을 모두 조사에 활용 될 수있다.

<table border="1">
<tbody>
 <tr>
 <td>Instrument/Drugs</td>
 <td>Company</td>
 <td>Cat #</td>
 <td>Comments</td>
 </tr>
 <tr>
 <td>Computer controlled impactor</td>
 <td>Leica or the Infinite Horizons (formerly OSU) impactor</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td></td>
 <td></td>
 <td></td>
 <td>Surgical instruments </td>
 </tr>
 <tr>
 <td>Scissors</td>
 <td>Fine Science Tools Inc
 </td>
 <td>14094-11 or 14060-09</td>
 <td></td>
 </tr>
 <tr>
 <td>Forceps</td>
 <td>Fine Science Tools Inc </td>
 <td>11006-12 and 11027-12 or 			11506-12</td>
 <td></td>
 </tr>
 <tr>
 <td>Hemostats</td>
 <td>Fine Science Tools Inc </td>
 <td>13009-12</td>
 <td></td>
 </tr>
 <tr>
 <td>Retractors</td>
 <td>Fine Science Tools Inc </td>
 <td>17011-10</td>
 <td></td>
 </tr>
 <tr>
 <td>Rongeurs</td>
 <td>Fine Science Tools Inc </td>
 <td>16020-14</td>
 <td></td>
 </tr>
 <tr>
 <td>Needle driver</td>
 <td>Fine Science Tools Inc </td>
 <td>12001-13</td>
 <td></td>
 </tr>
 <tr>
 <td>Stereotactic frame</td>
 <td> Leica or RWD Life Science Co. or
 TSE systems</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Buprinorphine</td>
 <td></td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Baytril</td>
 <td>Bayer</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Ketamine</td>
 <td></td>
 <td></td>
 <td></td>
 </tr>
 </tbody>
</table>

a contusion model of severe spinal cord injury in rats

새로운 치료의 번역 가능성은 심각한 척수 손상 (SCI) 타박상 모델에서 조사해야한다. 자세한 방법론은 심각한 SCI의 일관된 모델을 얻기 위해 설명되어 있습니다. 정위 구조 및 컴퓨터 제어 충격기의 사용은 재현성 부상을 생성 할 수 있습니다. 저체온증과 요로 감염은 수술 후 기간에 상당한 도전을 제기. 일일 체중 기록 및 방광 식 동물의주의 깊은 모니터링이 수술 후 합병증의 조기 발견 할 수 있습니다. 이 타박상 모델의 기능적 결과를 절개 모델과 동일합니다. 타박상 모델은 신경과 neuroregenerative 방법 모두의 효능을 평가하기 위하여 이용 될 수있다.

병변 볼륨 우리는 기술은 위에 설명에 따라 크고 일관된 병변 볼륨을 얻을 수있다. 2.04 mm 3 (1.9-2.18 CI 95 %) (N = 5 동물)를 얻었다의 평균 병변 볼륨을 염색 Luxol 빠른 파란색. 그림 2를 사용하여 병변 진원지를 통해 Luxol 빠른 블루를 사용하는 대표적인 염색 병변 볼륨을 의미합니다. 기능 점수 바소, 비티, 브레스 (BBB) ​...

Watch this Scientific Journal Video about A Contusion Model of Severe Spinal Cord Injury in Rats at JoVE.com

A Contusion Model of Severe Spinal Cord Injury in Rats

심각한 척수 손상의 타박상 모델이 설명되어 있습니다. 자세한 수술 전, 수술 및 수술 후 단계는 일관된 모델을 얻기 위해 설명되어 있습니다.

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-contusion-model-of-severe-spinal-cord-injury-in-rats

Research

JoVE Journal

Medicine

30.2K 조회수.  Medical University of South Carolina. 심각한 척수 손상의 타박상 모델이 설명되어 있습니다. 자세한 수술 전, 수술 및 수술 후 단계는 일관된 모델을 얻기 위해 설명되어 있습니다.

비디오: A Contusion Model of Severe Spinal Cord Injury in 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 ...

연구

의학

A Contusive Model of Unilateral Cervical Spinal Cord Injury Using the Infinite Horizon Impactor

While the majority of human spinal cord injuries occur in the cervical spinal cord, the vast majority of laboratory research employs animal models of spinal cord injury (SCI) in which the thoracic spinal cord is injured. Additionally, because most human cord injuries occur as the result of blunt, non-penetrating trauma (e.g. motor vehicle accident, sporting injury) where the spinal cord is violently struck by displaced bone or soft tissues, the majority of SCI researchers are of the opinion that the most clinically relevant injury models are those in which the spinal cord is rapidly contused.1 Therefore, an important step in the preclinical evaluation of novel treatments on their way to human translation is an assessment of their efficacy in a model of contusion SCI within the cervical spinal cord. Here, we describe the technical aspects and resultant anatomical and behavioral outcomes of an unilateral contusive model of cervical SCI that employs the Infinite Horizon spinal cord injury impactor.
Sprague Dawley rats underwent a left-sided unilateral laminectomy at C5. To optimize the reproducibility of the biomechanical, functional, and histological outcomes of the injury model, we contused the spinal cords using an impact force of 150 kdyn, an impact trajectory of 22.5&deg; (animals rotated at 22.5&deg;), and an impact location off of midline of 1.4 mm. Functional recovery was assessed using the cylinder rearing test, horizontal ladder test, grooming test and modified Montoya's staircase test for up to 6 weeks, after which the spinal cords were evaluated histologically for white and grey matter sparing.
The injury model presented here imparts consistent and reproducible biomechanical forces to the spinal cord, an important feature of any experimental SCI model. This results in discrete histological damage to the lateral half of the spinal cord which is largely contained to the ipsilateral side of injury. The injury is well tolerated by the animals, but does result in functional deficits of the forelimb that are significant and sustained in the weeks following injury. The cervical unilateral injury model presented here may be a resource to researchers who wish to evaluate potentially promising therapies prior to human translation.

A reliable and repeatable way to produce a cervical unilateral spinal cord injury using the Infinite Horizon impactor is described. The method takes advantage of a custom designed frame and clamp to stabilize the spine. The standardized procedure and biomechanical injury parameters result in sufficient and sustained injuries.

무한 호라이즌 임팩터을 사용하여 일방적 경부 척수의 부상의 Contusive 모델

While the majority of human spinal cord injuries occur in the cervical spinal cord, the vast majority of laboratory research employs animal models of ...

A Murine Model of Cervical Spinal Cord Injury to Study Post-lesional Respiratory Neuroplasticity

A cervical spinal cord injury induces permanent paralysis, and often leads to respiratory distress. To date, no efficient therapeutics have been developed to improve/ameliorate the respiratory failure following high cervical spinal cord injury (SCI). Here we propose a murine pre-clinical model of high SCI at the cervical 2 (C2) metameric level to study diverse post-lesional respiratory neuroplasticity. The technique consists of a surgical partial injury at the C2 level, which will induce a hemiparalysis of the diaphragm due to a deafferentation of the phrenic motoneurons from the respiratory centers located in the brainstem. The contralateral side of the injury remains intact and allows the animal recovery. Unlike other SCIs which affect the locomotor function (at the thoracic and lumbar level), the respiratory function does not require animal motivation and the quantification of the deficit/recovery can be easily performed (diaphragm and phrenic nerve recordings, whole body ventilation). This pre-clinical C2 SCI model is a powerful, useful, and reliable pre-clinical model to study various respiratory and non-respiratory neuroplasticity events at different levels (molecular to physiology) and to test diverse putative therapeutic strategies which might improve the respiration in SCI patients.

Respiratory failure is the leading cause of death following a cervical spinal cord injury. Having a reproducible, quantifiable, and reliable pre-clinical animal model of respiratory failure induced by a partial cervical injury will help to understand the subsequent respiratory and non-respiratory neuroplasticity and allow testing putative repair strategies.

후 병변 호흡 신경 가소성을 연구하기 위해 자궁 경부 척수 손상의 생쥐 모델

A cervical spinal cord injury induces permanent paralysis, and often leads to respiratory distress. To date, no efficient therapeutics have been developed ...

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, ...

A Radio-telemetric System to Monitor Cardiovascular Function in Rats with Spinal Cord Transection and Embryonic Neural Stem Cell Grafts

High thoracic or cervical spinal cord injury (SCI) can lead to cardiovascular dysfunction. To monitor cardiovascular parameters, we implanted a catheter connected to a radio transmitter into the femoral artery of rats that underwent a T4 spinal cord transection with or without grafting of embryonic brainstem-derived neural stem cells expressing green fluorescent protein. Compared to other methods such as cannula insertion or tail-cuff, telemetry is advantageous to continuously monitor blood pressure and heart rate in freely moving animals. It is also capable of long term multiple data acquisitions. In spinal cord injured rats, basal cardiovascular data under unrestrained condition and autonomic dysreflexia in response to colorectal distension were successfully recorded. In addition, cardiovascular parameters before and after SCI can be compared in the same rat if a transmitter is implanted before a spinal cord transection. One limitation of the described telemetry procedure is that implantation in the femoral artery may influence the blood supply to the ipsilateral hindlimb.

We present a protocol for using a radio-telemetric system to record cardiovascular parameters in T4 spinal cord transected rats eight weeks after embryonic brainstem neural stem cell grafting into the lesion site. Telemetry is an advanced technique to accurately evaluate cardiovascular function in conscious freely moving spinal cord injured rats.

무선 원격 측정 시스템은 척수 절개 및 배아 신경 세포 이식 줄기와 흰쥐의 심장 혈관 기능을 모니터링 할 수

High thoracic or cervical spinal cord injury (SCI) can lead to cardiovascular dysfunction. To monitor cardiovascular parameters, we implanted a catheter ...

Neural Stem Cell Transplantation in Experimental Contusive Model of Spinal Cord Injury

Spinal cord injury is a devastating clinical condition, characterized by a complex of neurological dysfunctions. Animal models of spinal cord injury can be used both to investigate the biological responses to injury and to test potential therapies. Contusion or compression injury delivered to the surgically exposed spinal cord are the most widely used models of the pathology. In this report the experimental contusion is performed by using the Infinite Horizon (IH) Impactor device, which allows the creation of a reproducible injury animal model through definition of specific injury parameters. Stem cell transplantation is commonly considered a potentially useful strategy for curing this debilitating condition. Numerous studies have evaluated the effects of transplanting a variety of stem cells. Here we demonstrate an adapted method for spinal cord injury followed by tail vein injection of cells in CD1 mice. In short, we provide procedures for: i) cell labeling with a vital tracer, ii) pre-operative care of mice, iii) execution of a contusive spinal cord injury, and iv) intravenous administration of post mortem neural precursors. This contusion model can be utilized to evaluate the efficacy and safety of stem cell transplantation in a regenerative medicine approach.

Spinal cord injury is a traumatic condition that causes severe morbidity and high mortality. In this work we describe in detail a contusion model of spinal cord injury in mice followed by a transplantation of neural stem cells.

척수 손상의 실험 Contusive 모델에서 신경 줄기 세포 이식

Spinal cord injury is a devastating clinical condition, characterized by a complex of neurological dysfunctions. Animal models of spinal cord injury can ...

Calibrated Forceps Model of Spinal Cord Compression Injury

Compression injuries of the murine spinal cord are valuable animal models for the study of spinal cord injury (SCI) and spinal regenerative therapy. The calibrated forceps model of compression injury is a convenient, low cost, and very reproducible animal model for SCI. We used a pair of modified forceps in accordance with the method published by Plemel et al. (2008) to laterally compress the spinal cord to a distance of 0.35 mm. In this video, we will demonstrate a dorsal laminectomy to expose the spinal cord, followed by compression of the spinal cord with the modified forceps. In the video, we will also address issues related to the care of paraplegic laboratory animals. This injury model produces mice that exhibit impairment in sensation, as well as impaired hindlimb locomotor function. Furthermore, this method of injury produces consistent aberrations in the pathology of the SCI, as determined by immunohistochemical methods. After watching this video, viewers should be able to determine the necessary supplies and methods for producing SCI of various severities in the mouse for studies on SCI and/or treatments designed to mitigate impairment after injury.

Spinal cord injury models should be highly reproducible. We demonstrate that the calibrated forceps compression model of spinal cord injury is an easy to use surgical method for generating reproducible injuries to the murine spinal cord.

척수 압축 부상의 보정 집게 모델

Compression injuries of the murine spinal cord are valuable animal models for the study of spinal cord injury (SCI) and spinal regenerative therapy. The ...

Contrast Enhanced Ultrasound Imaging for Assessment of Spinal Cord Blood Flow in Experimental Spinal Cord Injury

Reduced spinal cord blood flow (SCBF) (i.e., ischemia) plays a key role in traumatic spinal cord injury (SCI) pathophysiology and is accordingly an important target for neuroprotective therapies. Although several techniques have been described to assess SCBF, they all have significant limitations. To overcome the latter, we propose the use of real-time contrast enhanced ultrasound imaging (CEU). Here we describe the application of this technique in a rat contusion model of SCI. A jugular catheter is first implanted for the repeated injection of contrast agent, a sodium chloride solution of sulphur hexafluoride encapsulated microbubbles. The spine is then stabilized with a custom-made 3D-frame and the spinal cord dura mater is exposed by a laminectomy at ThIX-ThXII. The ultrasound probe is then positioned at the posterior aspect of the dura mater (coated with ultrasound gel). To assess baseline SCBF, a single intravenous injection (400 &#181;l) of contrast agent is applied to record its passage through the intact spinal cord microvasculature. A weight-drop device is subsequently used to generate a reproducible experimental contusion model of SCI. Contrast agent is re-injected 15 min following the injury to assess post-SCI SCBF changes. CEU allows for real time and in-vivo assessment of SCBF changes following SCI. In the uninjured animal, ultrasound imaging showed uneven blood flow along the intact spinal cord. Furthermore, 15 min post-SCI, there was critical ischemia at the level of the epicenter while SCBF remained preserved in the more remote intact areas. In the regions adjacent to the epicenter (both rostral and caudal), SCBF was significantly reduced. This corresponds to the previously described &#8220;ischemic penumbra zone&#8221;. This tool is of major interest for assessing the effects of therapies aimed at limiting ischemia and the resulting tissue necrosis subsequent to SCI.

Contrast Enhanced Ultrasound imaging is a reliable in-vivo tool for quantifying spinal cord blood flow in an experimental rat spinal cord injury model. This paper contains a comprehensive protocol for application of this technique in association with a contusion model of thoracic spinal cord injury.

실험 척수 부상에서 척수 혈액 흐름의 평가를위한 향상된 초음파 이미징 대비

Reduced spinal cord blood flow (SCBF) (i.e., ischemia) plays a key role in traumatic spinal cord injury (SCI) pathophysiology and is accordingly an ...

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) ...

A Neonatal Mouse Spinal Cord Compression Injury Model

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal cord. A major current area of research is focused on the mechanisms of adaptive plasticity that underlie spontaneous or induced functional recovery following SCI. Spontaneous functional recovery is reported to be greater early in life, raising interesting questions about how adaptive plasticity changes as the spinal cord develops. To facilitate investigation of this dynamic, we have developed a SCI model in the neonatal mouse. The model has relevance for pediatric SCI, which is too little studied. Because neural plasticity in the adult involves some of the same mechanisms as neural plasticity in early life1, this model may potentially have some relevance also for adult SCI. Here we describe the entire procedure for generating a reproducible spinal cord compression (SCC) injury in the neonatal mouse as early as postnatal (P) day 1. SCC is achieved by performing a laminectomy at a given spinal level (here described at thoracic levels 9-11) and then using a modified Yasargil aneurysm mini-clip to rapidly compress and decompress the spinal cord. As previously described, the injured neonatal mice can be tested for behavioral deficits or sacrificed for ex vivo physiological analysis of synaptic connectivity using electrophysiological and high-throughput optical recording techniques1. Earlier and ongoing studies using behavioral and physiological assessment have demonstrated a dramatic, acute impairment of hindlimb motility followed by a complete functional recovery within 2 weeks, and the first evidence of changes in functional circuitry at the level of identified descending synaptic connections1.

This article describes a method for generating a reproducible spinal cord compression injury (SCI) in the neonatal mouse. The model provides an advantageous platform for studying mechanisms of adaptive plasticity that underlie spontaneous functional recovery.

신생아 마우스 척수 압축 부상 모델

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal ...