Este trabalho é apoiado pelo financiamento do programa-quadro da União Europeia Sétimo (FP7/2007-2013), sob contrato de subvenção n ° 246556 (projeto RBUCE-UP Europeia), HandiMedEx atribuído pelo Conselho de Investimento Público francês. Marcel Bonay foi apoiado pelo Chancellerie des Universidades de Paris (Pernas Poix), o Fonds de Recherche en Santé Dotation respiratoire, eo Centre d&#39;Assistência respiratoire domicílio d&#39;Île de France (CARDIF)

Este protocolo foi aprovado pelo comitê de ética da cadeira RBUCE-UP de Excelência (Universidade de Paris Sud, convenção de subvenção n º 246556) e da Université de Versailles Saint-Quentin-en-Yvelines. 1. Preparação de instrumentos cirúrgicos esterilizados <ol><li> Limpe os instrumentos cirúrgicos com detergente de laboratório. </li><li> Autoclave os instrumentos antes da cirurgia. </li><li> Numa sessão cirúrgica, esterilizar as ferramentas, colocando as pontas de um esterilizador talão quente durante 10 minutos a 180 ° C entre as duas operações. </li></ol> 2. Preparação de Drogas <ol><li> Prepare 2 x 1 ml seringas para drogas pré-anestésica drogas cocktail e pós-operação. </li><li> De acordo com o peso do rato, preparar uma seringa com medicamentos pré-anestésicos: carprofeno (5 mg / kg), buprenorfina (50 ug / kg), Baytril (5 mg / kg) e dexmedetomidina (0,5 mg / kg). Completar o volume para 1 ml com solução de Ringer. </li><li> Prepare-seuma outra seringa para a reversão das drogas pré-anestésicos: atipamezol (500 ug / kg). </li></ol> 3. Anestesia do Rato <ol><li> Administrar por via subcutânea ao animal, a solução de drogas pré-anestésicos descritos no passo 2.2. Em seguida, coloque o animal de volta na gaiola e espere até que o efeito sedativo aparece. </li><li> Coloque o rato em uma câmara fechada preenchida com 5% de isoflurano em 100% O 2, e esperar até que o ritmo respiratório diminui (cerca de 30 seg.) Em seguida, retire o rato da câmara e coloque-a sobre a mesa de intubação. </li></ol> 4. Intubação orotraqueal <ol><li> Deite o animal de costas, em seguida, segura a cabeça, colocando uma alça ligada a seus dentes da frente para a mesa. </li><li> Com uma luz de fibra óptica, iluminar o espaço torácico. Em seguida, coloque um laringoscópio (ou uma custom made ​​um, Jou et al. 20 para mais detalhes) na boca do animal. Visualize as cordas vocais. </li><li> Deslizoue e colocar um guia orotraqueal na traqueia (entre as cordas vocais). Deslize o tubo orotraqueal (16 G tamanho do cateter) na guia. </li><li> Remover o guia e verificar com um espelho de laringe colocado na extremidade do tubo orotraqueal para a presença de humidade, o que confirma a posição apropriada do tubo na traqueia e não no esófago. </li><li> Ligar o tubo de um ventilador de roedor (683 roedor ventilador, Harvard Apparatus) e ajustar a concentração de isoflurano a 2% (em 100% de O2). </li><li> Fixe o tubo orotraqueal com esparadrapo. </li></ol> 5. Cirurgia da Coluna Vertebral <ol><li> Colocar o animal em decúbito ventral em uma placa cirúrgica aquecida, com o nariz apontando para ângulo de 90 ° com o cirurgião. Manter a temperatura do corpo de cerca de 37,5 ° C durante a cirurgia. </li><li> Raspar o cabelo com tesouras entre as escápulas e remover o cabelo com uma gaze. </li><li> Limpar a pele com betadine e em seguida com 70% de álcool. Repita este passo 3x. </li><li> Uma pitada do dedo do pé é realizada antes do início da cirurgia para garantir profundidade anestésica adequada. &#160; Em seguida, faça uma incisão na pele rostro-caudal lateral com tesoura entre as escápulas. </li><li> Corte o músculo acromiotrapezius rostro-caudal, seguindo o tendão para evitar qualquer sangramento. Então dissociar o músculo rombóide para acessar os músculos spinalis (em torno da vértebra). </li><li> Recolha o músculo spinalis de C1 a C3 vértebra. A vértebra C2 é o único com uma apófise proeminente. </li><li> Limpe o músculo ao redor da parte dorsal da vértebra usando cotonetes estéreis. </li><li> Comece para remover cuidadosamente a apófise do C2 com um rongeur. Em seguida, continue meticulosamente até o dorsal da medula espinhal está exposto. Certifique-se que laminectomia é uma laminectomia hemi dorsal. Preste muita atenção para a dura-máter que envolve a medula espinhal e as artérias nas imediações da área. </li><li> Com # 55 fórceps, dissecar rostro-caudal da dura juntamente C2, ao lado continuam lateralmente de cada lado rostral e caudal. </li><li> Esponja o líquido cefalorraquidiano. </li><li> Faça uma secção lateral sob o número cervical da raiz dorsal 2 com a microtesoura. Verifique com um bisturi micro que a extensão da lesão é perto o suficiente para chegar à linha média da medula espinhal (ver Figura 2A para uma vista dorsal da lesão). Se não, então um outro corte pode ser feito para completar a lesão. Em caso de hemorragia, use cotonetes estéreis. Tenha cuidado para não ir para o lado contralateral, caso contrário, o animal não vai se recuperar da lesão e terá uma insuficiência respiratória. </li><li> Suturar os músculos como uma camada protetora e sutura de volta a pele. Limpe o ferimento com gaze esterilizada Betadine saturada. </li><li> Vire-off o vaporizador isoflurano e injetar as drogas Reversão (Atipamezole [500 mg / kg, im]), verifique a temperatura do corpo. </li><li> Quando o animal começa a respirar contra o ventilador,desconectar o tubo traqueal do ventilador, em seguida, remover o tubo orotraqueal. Colocar o animal numa gaiola aquecida para recuperação. </li></ol> 6. Cuidados pós-cirúrgica Após a cirurgia, os animais são monitorados continuamente para garantir o melhor ambiente possível para a recuperação. Antibióticos (Baytril, 5 mg / kg), anti-inflamatório (Carprofeno, 5 mg / kg) e buprenorfina (50 ug / kg) as drogas são dadas a cada 12 horas durante os primeiros dois dias pós-cirurgia para evitar infecções e reduzir a ocorrência da dor pós-operatória. Os ratos têm acesso ad libitum ao alimento macio e água (ou água rebuçados em gel para o 1 º dia de pós-operatório). Fluídos subcutâneos pode ser usado para evitar a desidratação nos primeiros dias de pós-operatório. O peso corporal e a ingestão de alimentos são monitorizados diariamente. Seu ambiente é enriquecido ao longo da experiência e do tempo pós-lesão (dual habitação, tubos em suas gaiolas).

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Dificuldades técnicas de fazer o ferimento Modelo C2 O modelo murino lesão C2 é uma ferramenta interessante para estudar respiratória neuroplasticidade pós-lesional. No entanto, os passos necessários para produzir um modelo reproduzível e confiável são numerosos e cada um pode ter um impacto sobre o resultado do estudo. Por exemplo, durante o processo de intubação, o maior cuidado deve ser tomado uma vez que o tubo orotraqueal pode produzir uma inflamação da tra...

Trauma medular é uma lesão comum observada na população humana com incidências dramáticas, tais como paralisia permanente. No entanto, a gravidade da lesão depende do grau e da extensão do trauma inicial. A insuficiência respiratória é a principal causa de mortalidade após lesão medular cervical superior (SCI) 1. Atualmente, o único tratamento terapêutico é colocar o paciente sob assistência ventilatória. Desde alguns pacientes podem ser desmamados a assistência ventilatória 2, devido à recuperação espontânea que ocorre com atraso pós-lesional, a necessidade de desenvolver novas terapias não-invasivas inovadores é urgente 3. Tendo um bom modelo pré-clínico normalizado para investigar o efeito de um SCI cervical em insuficiência respiratória e por isso, para estudar a aplicação de estratégias terapêuticas putativos, é essencial. Neste artigo técnico, nós descrevemos um modelo murino pré-clínico o específicocomprometimento respiratório f induzida por um SCI cervical parcial no nível C2. Este modelo é usado atualmente por vários laboratórios ao redor do mundo (para comentários: 4-13). No entanto, ligeiras diferenças de procedimento cirúrgico pode ser observada entre os diferentes pesquisadores para gerar este modelo murino lesão cervical particular. O efeito de um C2 SCI na saída respiratória foi descrita pela primeira vez em 1895 por Porter 14. Uma hemisecção cervical induz uma deaferentação dos motoneurónios frênicas da sua unidade central (localizado no rVRG no tronco cerebral, Figura 1A) no lado ipsilateral de lesão, levando a uma actividade do nervo frénico e silenciosa, o diafragma posterior paralisia. O lado contralateral permanece intacta e que permite ao animal sobreviver. Ao contrário diferente SCI localizado num segmento da coluna vertebral inferior (por exemplo, uma lesão contundente ao nível C4 15), a integridade do núcleo de motoneurónios frénico em ambos os lados é preservada. Após um CERVlesão C2 iCal, alguma atividade espontânea pode ser observado no lado ipsilateral (frênico e diafragma), devido a uma ativação de vias sinápticas silenciosas contralateral que cruzaram a linha média da coluna vertebral ao nível segmentar C3-C6 (caminhos cruzados frênico, CPP, Figura 1B) . A ativação do CPP, que é, por definição, um hemissecção C2 combinada com uma phrenicotomy contralateral que induzir uma recuperação parcial do nervo frênico ipsilateral, pode ocorrer de horas a semanas pós-lesão 16-18. O efeito benéfico real deste caminho CPP na recuperação respiratória é limitada 19 e mais investigação e tratamento devem ser desenvolvidos para melhorar a magnitude da recuperação espontânea 3. Este protocolo fornece um tipo poderoso de modelo murino pré-clínico para estudar a plasticidade pós-lesional respiratória em vários níveis (fisiologia respiratória de motoneurônios pré e frênico, interneurônios, molecular e cellular, a locomoção do membro dianteiro, por exemplo), bem como um modelo para testar as estratégias terapêuticas invasivas e não invasivas destinados a melhorar os sistemas respiratório e recuperação locomotora seguinte C2 lesão medular cervical parcial.

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			<td>225-250g</td>
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			<td height="21" style="height:21px;">Mayo-Stille Scissors</td>
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			<td height="21" style="height:21px;">Spring Scissors - 8 mm Blades</td>
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			<td>15025-10</td>
			<td>Straight Blunt/Blunt</td>
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			<td>Curved</td>
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			<td height="21" style="height:21px;">Dissecting Knife - Fine Tip</td>
			<td>Fine Science Tool</td>
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			<td>Straight</td>
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			<td>2x3 Blunt</td>
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			<td height="21" style="height:21px;">Absorbable surgical sutures</td>
			<td>Centravet</td>
			<td>BYO001</td>
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			<td height="21" style="height:21px;">Hot Bead Steriliser</td>
			<td>Fine Science Tool</td>
			<td>18000-45</td>
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			<td height="21" style="height:21px;">Catheter&#160;</td>
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			<td>16 gauge</td>
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			<td>Centravet</td>
			<td>MIR011</td>
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			<td height="21" style="height:21px;">Lactated Ringers</td>
			<td>Centravet</td>
			<td>RIN020</td>
			<td></td>
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			<td height="21" style="height:21px;">Syringe</td>
			<td>Centravet</td>
			<td></td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">Needle</td>
			<td>Centravet</td>
			<td></td>
			<td></td>
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			<td height="25" style="height:25px;">O2</td>
			<td>Air Liquid</td>
			<td>I1001M20R2A001</td>
			<td></td>
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			<td height="21" style="height:21px;">683 RodentT Ventilator 115/230V</td>
			<td>Havard Apparatus</td>
			<td>55-0000</td>
			<td></td>
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			<td height="21" style="height:21px;">Stand-Alone Vaporizer</td>
			<td>WPI</td>
			<td>EZ-155</td>
			<td></td>
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			<td height="21" style="height:21px;">Thin line heated bed</td>
			<td>WPI</td>
			<td>EZ-211</td>
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			<td>WPI</td>
			<td>EZ-258</td>
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			<td>Centravet</td>
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			<td>Centravet</td>
			<td>RIM011</td>
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			<td>BUP001</td>
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			<td>VET002</td>
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			<td height="21" style="height:21px;">Isoflurane</td>
			<td>Centravet</td>
			<td>VET066</td>
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a murine model of cervical spinal cord injury to study post-lesional respiratory neuroplasticity

A lesão da medula espinhal cervical induz a paralisia permanente, e muitas vezes leva a problemas respiratórios. Até o momento, não houve terapêutica eficientes foram desenvolvidos para melhorar / amenizar a insuficiência respiratória após alta lesão medular cervical (SCI). Aqui propomos um modelo pré-clínico murino de alta SCI na cervical 2 (C2) nível metamérica estudar diversificada neuroplasticidade respiratória pós-lesional. A técnica consiste numa lesão parcial cirúrgica ao nível C2, que irá induzir uma hemiparalisia do diafragma devido a uma deaferentação dos motoneurónios frênicas dos centros respiratórios localizados no tronco cerebral. O lado contralateral da lesão permanece intacta e permite a recuperação dos animais. Ao contrário de outros SIC que afetam a função locomotora (no tórax e nível lombar), a função respiratória não requer motivação animal e à quantificação do déficit / recuperação pode ser realizada facilmente (diafragma e gravação do nervo frênicos, ventilação de todo o corpo). Este modelo pré-clínico C2 SCI é um modelo pré-clínico poderoso, útil e confiável para estudar vários eventos neuroplasticidade respiratórios e não respiratórios em diferentes níveis (molecular) para a fisiologia e para testar diferentes estratégias terapêuticas putativos que possam melhorar a respiração em pacientes com LM.

Extensão da lesão O sucesso ea reprodutibilidade deste modelo experimental específico dependem da experiência de cada manipulador / cirurgião. A quantidade subseqüente de recuperação respiratória (atividade do nervo frênico e atividade do diafragma), após uma lesão no C2 está correlacionada com o restante ventrolateral poupado substância branca 21. Uma vez que a lesão é &quot;handmade&quot; e requer um pouco de prática do cirurgião, a extensão d...

Watch this Scientific Journal Video about A Murine Model of Cervical Spinal Cord Injury to Study Post-lesional Respiratory Neuroplasticity at JoVE.com

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

A insuficiência respiratória é a principal causa de morte na sequência de uma lesão medular cervical. Ter um modelo animal pré-clínico reprodutível, quantificáveis ​​e confiável de insuficiência respiratória induzida por uma lesão cervical parcial vai ajudar a entender a neuroplasticidade respiratório e não-respiratório posterior e permite testar estratégias de reparo putativos.

Um modelo murino de Spinal Cord Injury do colo do útero para o Estudo de pós-lesional respiratória Neuroplasticidade

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

a-murine-model-cervical-spinal-cord-injury-to-study-post-lesional

Research

JoVE Journal

Medicine

13.4K visualizações.  Université de Versailles Saint-Quentin-en-Yvelines.  A insuficiência respiratória é a principal causa de morte na sequência de uma lesão medular cervical. Ter um modelo animal pré-clínico reprodutível, quantificáveis ​​e confiável de insuficiência respiratória induzida por uma lesão cervical parcial vai ajudar a entender a neuroplasticidade respiratório e não-respiratório posterior e permite testar estratégias de reparo putativos. 

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

Intraspinal Cell Transplantation for Targeting Cervical Ventral Horn in Amyotrophic Lateral Sclerosis and Traumatic Spinal Cord Injury

Respiratory compromise due to phrenic motor neuron loss is a debilitating consequence of a large proportion of human traumatic spinal cord injury (SCI) cases 1 and is the ultimate cause of death in patients with the motor neuron disorder, amyotrophic laterals sclerosis (ALS) 2.
ALS is a devastating neurological disorder that is characterized by relatively rapid degeneration of upper and lower motor neurons. Patients ultimately succumb to the disease on average 2-5 years following diagnosis because of respiratory paralysis due to loss of phrenic motor neuron innnervation of the diaphragm 3. The vast majority of cases are sporadic, while 10% are of the familial form. Approximately twenty percent of familial cases are linked to various point mutations in the Cu/Zn superoxide dismutase 1 (SOD1) gene on chromosome 21 4. Transgenic mice 4,5 and rats 6 carrying mutant human SOD1 genes (G93A, G37R, G86R, G85R) have been generated, and, despite the existence of other animal models of motor neuron loss, are currently the most highly used models of the disease.
Spinal cord injury (SCI) is a heterogeneous set of conditions resulting from physical trauma to the spinal cord, with functional outcome varying according to the type, location and severity of the injury 7. Nevertheless, approximately half of human SCI cases affect cervical regions, resulting in debilitating respiratory dysfunction due to phrenic motor neuron loss and injury to descending bulbospinal respiratory axons 1. A number of animal models of SCI have been developed, with the most commonly used and clinically-relevant being the contusion 8.
Transplantation of various classes of neural precursor cells (NPCs) is a promising therapeutic strategy for treatment of traumatic CNS injuries and neurodegeneration, including ALS and SCI, because of the ability to replace lost or dysfunctional CNS cell types, provide neuroprotection, and deliver gene factors of interest 9.
Animal models of both ALS and SCI can model many clinically-relevant aspects of these diseases, including phrenic motor neuron loss and consequent respiratory compromise 10,11. In order to evaluate the efficacy of NPC-based strategies on respiratory function in these animal models of ALS and SCI, cellular interventions must be specifically directed to regions containing therapeutically relevant targets such as phrenic motor neurons. We provide a detailed protocol for multi-segmental, intraspinal transplantation of NPCs into the cervical spinal cord ventral gray matter of neurodegenerative models such as SOD1G93A mice and rats, as well as spinal cord injured rats and mice 11.

Neural precursor transplantation is a promising strategy for protecting and/or replacing lost/dysfunctional cervical phrenic motor neurons in spinal cord injury (SCI) and the motor neuron disorder, amyotrophic laterals sclerosis (ALS). We provide a protocol for cell delivery to cervical spinal cord ventral horn in rodent models of ALS and SCI.

Intra Transplante de Células de Segmentação Chifre Cervical Ventral em Esclerose Lateral Amiotrófica e lesão medular traumática

Respiratory compromise due to phrenic motor neuron loss is a debilitating consequence of a large proportion of human traumatic spinal cord injury (SCI) ...

Medicina

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.

Aguda e crônica teste sensorial tátil após a lesão medular em ratos

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

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.

Um Modelo contundente de lesão da medula espinhal cervical unilateral Usando o Impactor horizonte infinito

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

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.

Controlada por laceração do colo do útero em camundongos

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.

Um modelo de contusão grave de lesão medular em ratos

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

Evaluation of Respiratory Muscle Activation Using Respiratory Motor Control Assessment (RMCA) in Individuals with Chronic Spinal Cord Injury

During breathing, activation of respiratory muscles is coordinated by integrated input from the brain, brainstem, and spinal cord. When this coordination is disrupted by spinal cord injury (SCI), control of respiratory muscles innervated below the injury level is compromised1,2 leading to respiratory muscle dysfunction and pulmonary complications. These conditions are among the leading causes of death in patients with SCI3. Standard pulmonary function tests that assess respiratory motor function include spirometrical and maximum airway pressure outcomes: Forced Vital Capacity (FVC), Forced Expiratory Volume in one second (FEV1), Maximal Inspiratory Pressure (PImax) and Maximal Expiratory Pressure (PEmax)4,5. These values provide indirect measurements of respiratory muscle performance6. In clinical practice and research, a surface electromyography (sEMG) recorded from respiratory muscles can be used to assess respiratory motor function and help to diagnose neuromuscular pathology. However, variability in the sEMG amplitude inhibits efforts to develop objective and direct measures of respiratory motor function6. Based on a multi-muscle sEMG approach to characterize motor control of limb muscles7, known as the voluntary response index (VRI)8, we developed an analytical tool to characterize respiratory motor control directly from sEMG data recorded from multiple respiratory muscles during the voluntary respiratory tasks. We have termed this the Respiratory Motor Control Assessment (RMCA)9. This vector analysis method quantifies the amount and distribution of activity across muscles and presents it in the form of an index that relates the degree to which sEMG output within a test-subject resembles that from a group of healthy (non-injured) controls. The resulting index value has been shown to have high face validity, sensitivity and specificity9-11. We showed previously9 that the RMCA outcomes significantly correlate with levels of SCI and pulmonary function measures. We are presenting here the method to quantitatively compare post-spinal cord injury respiratory multi-muscle activation patterns to those of healthy individuals.

Evaluation of Respiratory Muscle Activation Using Respiratory Motor Control Assessment RMCA in Individuals with Chronic Spinal Cord Injury

The purpose of this publication is to present our original work on a multi-muscle surface electromyographic approach to quantitatively characterize respiratory muscle activation patterns in individuals with chronic spinal cord injury using vector-based analysis.

Avaliação da ativação muscular respiratória Usando respiratória Motor Control Assessment (ACMD) em indivíduos com lesão medular crônica

During breathing, activation of respiratory muscles is coordinated by integrated input from the brain, brainstem, and spinal cord. When this coordination ...

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.

Controlado Modelo Impact Cortical para lesão cerebral traumática

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.

Um sistema de rádio-telemetria para monitorar a função cardiovascular em ratos com Medula Espinhal transecção e Embryonic Stem Neural enxertos de células-

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

Synergetic Use of Neural Precursor Cells and Self-assembling Peptides in Experimental Cervical Spinal Cord Injury

Spinal cord injuries (SCI) cause serious neurological impairment and psychological, economic, and social consequences for patients and their families. Clinically, more than 50% of SCI affect the cervical spine1. As a consequence of the primary injury, a cascade of secondary mechanisms including inflammation, apoptosis, and demyelination occur finally leading to tissue scarring and development of intramedullary cavities2,3. Both represent physical and chemical barriers to cell transplantation, integration, and regeneration. Therefore, shaping the inhibitory environment and bridging cavities to create a supportive milieu for cell transplantation and regeneration is a promising therapeutic target4. Here, a contusion/compression model of cervical SCI using an aneurysm clip is described. This model is more clinically relevant than other experimental models, since complete transection or ruptures of the cord are rare. Also in comparison to the weight drop model, which in particular damage the dorsum columns, circumferential compression of the spinal cord appears advantageous. Clip closing force and duration can be adjusted to achieve different injury severity. A ring spring facilitates precise calibration and constancy of clip force. Under physiological conditions, synthetic self-assembling peptides (SAP) self-assemble into nanofibers and thus, are appealing for application in SCI5. They can be injected directly into the lesion minimizing damage to the cord. SAPs are biocompatible structures erecting scaffolds to bridge intramedullary cavities and thus, equip the damaged cord for regenerative treatments. K2(QL)6K2 (QL6) is a novel SAP introduced by Dong et al.6 In comparison to other peptides, QL6 self-assembles into &#946;-sheets at neutral pH6.14 days after SCI, after the acute stage, SAPs are injected into the center of the lesion and neural precursor cells (NPC) are injected into adjacent dorsal columns. In order to support cell survival, transplantation is combined with continuous subdural administration of growth factors by osmotic micro pumps for 7 days.

Treating cervical spinal cord injury with both self-assembling peptides (SAP) and neural precursor cells (NPC), together with growth factors, is a promising approach to promote regeneration and recovery. A contusion/compression aneurysm clip rat model of cervical SCI and combined treatment involving SAP injection and NPC transplantation is established.

Use Synergetic de células neuronais precursoras e Peptídeos de auto-montagem em Experimental Cervical Spinal Cord Injury

Spinal cord injuries (SCI) cause serious neurological impairment and psychological, economic, and social consequences for patients and their families. ...

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.

Calibrado Fórceps Modelo de Lesão Medular Compression

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