Questo lavoro è supportato da un finanziamento del Programma quadro dell&#39;Unione europea Settimo (FP7/2007-2013) in convenzione di sovvenzione n ° 246.556 (progetto RBUCE-UP europeo), HandiMedEx assegnato dai francesi consiglio investimenti pubblici. Marcel Bonay è stato sostenuto dalla Cancelleria des Universités de Paris (Legs Poix), il Fonds de Recherche en Santé Dotation Respiratoire, e il Centro d&#39;Assistenza Respiratoire à Domicile d&#39;Île de France (CARDIF)

Questo protocollo è stato approvato dal comitato etico della sedia RBUCE-UP di eccellenza (Università di Paris Sud, convenzione di sovvenzione n ° 246.556) e l&#39;Université de Versailles Saint-Quentin-en-Yvelines. 1. Preparazione di strumenti chirurgici sterilizzati <ol><li> Pulire gli strumenti chirurgici con detergente da laboratorio. </li><li> Sterilizzare gli strumenti prima della chirurgia. </li><li> In una sessione chirurgica, sterilizzare gli strumenti posizionando le punte in un cordone sterilizzatore caldo per 10 min a 180 ° C tra 2 interventi chirurgici. </li></ol> 2. Preparazione dei farmaci <ol><li> Preparare 2 x 1 ml siringhe per i farmaci pre-anestetico cocktail e post-operazione droghe. </li><li> Secondo il peso del ratto, preparare una siringa con farmaci pre-anestetici: carprofen (5 mg / kg), buprenorfina (50 mg / kg), Baytril (5 mg / kg) e Dexmedetomidine (0,5 mg / kg). Completa il volume a 1 ml con Ringer lattato. </li><li> Preparare inun&#39;altra siringa l&#39;inversione per i farmaci pre-anestetico: atipamezolo (500 mcg / kg). </li></ol> 3. Anestesia del Topo <ol><li> Somministrare per via sottocutanea per l&#39;animale la soluzione dei farmaci pre-anestetico di cui al punto 2.2. Poi, mettere l&#39;animale torna in gabbia e aspettare che l&#39;effetto sedativo. </li><li> Posizionare il topo in una camera chiusa riempita di 5% isoflurano in 100% O 2, e attendere che il ritmo respiratorio rallenta (circa 30 sec). Quindi, rimuovere il ratto dalla camera e posizionarlo sul tavolo intubazione. </li></ol> 4. Orotracheale intubazione <ol><li> Lie l&#39;animale sulla schiena, quindi fissare la testa mettendo una cinghia collegata a denti davanti al tavolo. </li><li> Con una luce in fibra ottica, illuminare lo spazio toracico. Poi, mettere un laringoscopio (o un costume fatto uno, Jou et al. 20 per i dettagli) nella bocca dell&#39;animale. Visualizza le corde vocali. </li><li> SlidE e posizionare una guida orotracheal nella trachea (tra le corde vocali). Far scorrere il tubo orotracheale (16 G dimensione del catetere) sulla guida. </li><li> Rimuovere la guida e controllare con uno specchio laringeo posto all&#39;estremità del tubo orotracheale per la presenza di umidità, confermando la posizione appropriata del tubo nella trachea e non nell&#39;esofago. </li><li> Collegare il tubo di un ventilatore roditore (683 roditore ventilatore, Harvard Apparatus) e regolare la concentrazione di isoflurano al 2% (100% O 2). </li><li> Fissare il tubo orotracheale con del nastro adesivo chirurgico. </li></ol> 5. Chirurgia Vertebrale <ol><li> Posto l&#39;animale in decubito ventrale su una placca chirurgica riscaldata, con il naso che punta a 90 ° per il chirurgo. Mantenere la temperatura corporea intorno 37,5 ° C per tutta la chirurgia. </li><li> Radersi i capelli con Clippers tra le scapole e rimuovere i capelli con una garza. </li><li> Pulire la pelle con betadine, poi con 70% di alcol. Ripetere questa operazione 3 volte. </li><li> Un pizzico piedi viene eseguita prima dell&#39;inizio della chirurgia per garantire la corretta profondità dell&#39;anestesia. &#160; Quindi, eseguire una laterale incisione cutanea rostro-caudale con le forbici tra le scapole. </li><li> Tagliare il acromiotrapezius muscolo rostro-caudale seguendo il tendine per evitare emorragie. Poi dissociare il muscolo romboide per accedere ai muscoli spinale (che circondano la vertebra). </li><li> Ritrarre il muscolo spinale da C1 a C3 vertebra. La vertebra C2 è quella con un apofisi prominente. </li><li> Pulire il muscolo intorno alla parte dorsale della vertebra utilizzando tamponi di cotone sterile. </li><li> Iniziare a rimuovere con attenzione l&#39;apofisi di C2 con un rongeur. Quindi, continuare meticolosamente fino alla dorsale del midollo spinale è esposto. Assicurarsi che laminectomia è una laminectomia emi dorsale. Prestare particolare attenzione alla dura madre che racchiude il midollo spinale e le arterie in prossimità di questa zona. </li><li> Con # 55 pinze, sezionare rostro-caudale la dura lungo C2, il prossimo proseguire lateralmente su ciascun lato rostrale e caudale. </li><li> Spugna il liquido cerebrospinale. </li><li> Effettuare una sezione laterale sotto la cervicale dorsale radice numero 2 con le microscissors. Verificare con un micro bisturi che l&#39;estensione della lesione è abbastanza vicino per raggiungere la linea mediana del midollo spinale (vedere la Figura 2A per una vista dorsale della lesione). Se no, allora un altro taglio potrebbe essere fatto per completare la lesione. In caso di sanguinamento, utilizzare tamponi di cotone sterili. Fare attenzione a non andare al lato controlaterale, in caso contrario, l&#39;animale non si riprenderà dall&#39;infortunio e avrà una insufficienza respiratoria. </li><li> Suturare i muscoli come strato protettivo e suturare la pelle. Pulire la ferita con Betadine saturo garza sterile. </li><li> Turn-off il vaporizzatore isoflurano e iniettare i farmaci di inversione (atipamezolo [500 mg / kg, im]), controllare la temperatura corporea. </li><li> Quando l&#39;animale inizia a respirare contro il ventilatore,scollegare il tubo tracheale dal ventilatore e quindi rimuovere il tubo orotracheale. Posto l&#39;animale in una gabbia riscaldata per il recupero. </li></ol> 6. Assistenza post-chirurgica Dopo l&#39;intervento chirurgico, gli animali sono costantemente monitorati per garantire il miglior ambiente possibile per il recupero. Antibiotici (Baytril, 5 mg / kg), anti-infiammatori (Carprofene, 5 mg / kg) e buprenorfina (50 mg / kg) farmaci sono somministrati ogni 12 ore per i primi 2 giorni post-intervento chirurgico per prevenire infezioni e ridurre il verificarsi del dolore post-operatorio. I ratti hanno accesso ad libitum a cibo morbido e acqua (o acqua gelatine per la prima giornata post-operatoria). Fluidi sottocutanei possono essere usati per prevenire la disidratazione sui primi giorni post-operatorie. Il peso corporeo e l&#39;assunzione di cibo vengono monitorati giornalmente. Il loro ambiente si arricchisce tutto l&#39;esperimento e il tempo di post-infortunio (dual abitazioni, tubi nelle loro gabbie).

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	<li>Frankel, H. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-term+survival+in+spinal+cord+injury:+a+fifty+year+investigation.">Long-term survival in spinal cord injury: a fifty year investigation.</a> Spinal Cord. 36, 266-274 (1998).</li><li>Ramer, M. S., Harper, G. P., Bradbury, E. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Progress+in+spinal+cord+research+-+a+refined+strategy+for+the+International+Spinal+Research+Trust.">Progress in spinal cord research - a refined strategy for the International Spinal Research Trust.</a> Spinal Cord. 38, 449-472 (2000).</li><li>Zimmer, M. B., Nantwi, K., Goshgarian, H. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+spinal+cord+injury+on+the+respiratory+system:+basic+research+and+current+clinical+treatment+options.">Effect of spinal cord injury on the respiratory system: basic research and current clinical treatment options.</a> J Spinal Cord Med. 30, 319-330 (2007).</li><li>Mantilla, C. B., Sieck, G. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuromuscular+adaptations+to+respiratory+muscle+inactivity.">Neuromuscular adaptations to respiratory muscle inactivity.</a> Respir Physiol Neurobiol. 169, 133-140 (2009).</li><li>Goshgarian, H. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+crossed+phrenic+phenomenon+and+recovery+of+function+following+spinal+cord+injury.">The crossed phrenic phenomenon and recovery of function following spinal cord injury.</a> Respir Physiol Neurobiol. 169, 85-93 (2009).</li><li>Nantwi, K. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recovery+of+respiratory+activity+after+C2+hemisection+(C2HS):+involvement+of+adenosinergic+mechanisms.">Recovery of respiratory activity after C2 hemisection (C2HS): involvement of adenosinergic mechanisms.</a> Respir Physiol Neurobiol. 169, 102-114 (2009).</li><li>Sandhu, M. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Respiratory+recovery+following+high+cervical+hemisection.">Respiratory recovery following high cervical hemisection.</a> Respir Physiol Neurobiol. 169, 94-101 (2009).</li><li>Lane, M. A., Lee, K. Z., Fuller, D. D., Reier, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spinal+circuitry+and+respiratory+recovery+following+spinal+cord+injury.">Spinal circuitry and respiratory recovery following spinal cord injury.</a> Respir Physiol Neurobiol. 169, 123-132 (2009).</li><li>Seeds, N. W., Akison, L., Minor, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+plasminogen+activator+in+spinal+cord+remodeling+after+spinal+cord+injury.">Role of plasminogen activator in spinal cord remodeling after spinal cord injury.</a> Respir Physiol Neurobiol. 169, 141-149 (2009).</li><li>Alilain, W. J., Horn, K. P., Hu, H., Dick, T. E., Silver, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+regeneration+of+respiratory+pathways+after+spinal+cord+injury.">Functional regeneration of respiratory pathways after spinal cord injury.</a> Nature. 475, 196-200 (2011).</li><li>Vinit, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cervical+spinal+cord+injuries+and+respiratory+insufficiency:+a+revolutionary+treatment.">Cervical spinal cord injuries and respiratory insufficiency: a revolutionary treatment.</a> Med Sci (Paris. 28, 33-36 (2012).</li><li>Kastner, A., Gauthier, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Are+rodents+an+appropriate+pre-clinical+model+for+treating+spinal+cord+injury?+Examples+from+the+respiratory+system).">Are rodents an appropriate pre-clinical model for treating spinal cord injury? Examples from the respiratory system).</a> Exp Neurol. 213, 249-256 (2008).</li><li>Vinit, S., Lovett-Barr, M. R., Mitchell, G. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intermittent+hypoxia+induces+functional+recovery+following+cervical+spinal+injury.">Intermittent hypoxia induces functional recovery following cervical spinal injury.</a> Physiol Neurobiol. 169, 210-217 (2009).</li><li>Porter, W. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Path+of+the+Respiratory+Impulse+from+the+Bulb+to+the+Phrenic+Nuclei.">The Path of the Respiratory Impulse from the Bulb to the Phrenic Nuclei.</a> J Physiol. 17, 455-485 .</li><li>Nicaise, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phrenic+motor+neuron+degeneration+compromises+phrenic+axonal+circuitry+and+diaphragm+activity+in+a+unilateral+cervical+contusion+model+of+spinal+cord+injury.">Phrenic motor neuron degeneration compromises phrenic axonal circuitry and diaphragm activity in a unilateral cervical contusion model of spinal cord injury.</a> Exp Neurol. 235, 539-552 (2012).</li><li>Goshgarian, H. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+crossed+phrenic+phenomenon:+a+model+for+plasticity+in+the+respiratory+pathways+following+spinal+cord+injury.">The crossed phrenic phenomenon: a model for plasticity in the respiratory pathways following spinal cord injury.</a> J Appl Physiol. 94, 795-810 (2003).</li><li>Vinit, S., Gauthier, P., Stamegna, J. C., Kastner, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High+cervical+lateral+spinal+cord+injury+results+in+long-term+ipsilateral+hemidiaphragm+paralysis.">High cervical lateral spinal cord injury results in long-term ipsilateral hemidiaphragm paralysis.</a> J Neurotrauma. 23, 1137-1146 (2006).</li><li>Fuller, D. D., Johnson, S. M., Johnson, R. A., Mitchell, G. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+cervical+spinal+sensory+denervation+reveals+ineffective+spinal+pathways+to+phrenic+motoneurons+in+the+rat.">Chronic cervical spinal sensory denervation reveals ineffective spinal pathways to phrenic motoneurons in the rat.</a> Neurosci Lett. 323, 25-28 (2002).</li><li>Dougherty, B. J., Lee, K. Z., Lane, M. A., Reier, P. J., Fuller, D. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contribution+of+the+spontaneous+crossed-phrenic+phenomenon+to+inspiratory+tidal+volume+in+spontaneously+breathing+rats.">Contribution of the spontaneous crossed-phrenic phenomenon to inspiratory tidal volume in spontaneously breathing rats.</a> J Appl Physiol. 112, 96-105 (2012).</li><li>Jou, I. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simplified+rat+intubation+using+a+new+oropharyngeal+intubation+wedge.">Simplified rat intubation using a new oropharyngeal intubation wedge.</a> J Appl Physiol. 89, 1766-1770 (2000).</li><li>Fuller, D. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Graded+unilateral+cervical+spinal+cord+injury+and+respiratory+motor+recovery.">Graded unilateral cervical spinal cord injury and respiratory motor recovery.</a> Respir Physiol Neurobiol. 165, 245-253 (2009).</li><li>Vinit, S., Windelborn, J. A., Mitchell, G. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lipopolysaccharide+attenuates+phrenic+long-term+facilitation+following+acute+intermittent+hypoxia.">Lipopolysaccharide attenuates phrenic long-term facilitation following acute intermittent hypoxia.</a> Respir Physiol Neurobiol. 176, 130-135 (2011).</li><li>Ahmad, F., Wang, M. Y., Levi, A. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hypothermia+for+Acute+Spinal+Cord+Injury-A+Review.">Hypothermia for Acute Spinal Cord Injury-A Review.</a> World Neurosurg. , (2013).</li><li>Lovett-Barr, M. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Repetitive+intermittent+hypoxia+induces+respiratory+and+somatic+motor+recovery+after+chronic+cervical+spinal+injury.">Repetitive intermittent hypoxia induces respiratory and somatic motor recovery after chronic cervical spinal injury.</a> J Neurosci. 32, 3591-3600 (2012).</li><li>Minor, K. H., Akison, L. K., Goshgarian, H. G., Seeds, N. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spinal+cord+injury-induced+plasticity+in+the+mouse--the+crossed+phrenic+phenomenon.">Spinal cord injury-induced plasticity in the mouse--the crossed phrenic phenomenon.</a> Exp Neurol. 200, 486-495 (2006).</li><li>Baussart, B., Stamegna, J. C., Polentes, J., Tadie, M., Gauthier, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+model+of+upper+cervical+spinal+contusion+inducing+a+persistent+unilateral+diaphragmatic+deficit+in+the+adult+rat.">A new model of upper cervical spinal contusion inducing a persistent unilateral diaphragmatic deficit in the adult rat.</a> Neurobiol Dis. 22, 562-574 (2006).</li><li>Golder, F. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Breathing+patterns+after+mid-cervical+spinal+contusion+in+rats.">Breathing patterns after mid-cervical spinal contusion in rats.</a> Exp Neurol. 231, 97-103 (2011).</li><li>Lane, M. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Respiratory+function+following+bilateral+mid-cervical+contusion+injury+in+the+adult+rat.">Respiratory function following bilateral mid-cervical contusion injury in the adult rat.</a> Exp Neurol. 235, 197-210 (2012).</li><li>Vinit, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Axotomized+bulbospinal+neurons+express+c-Jun+after+cervical+spinal+cord+injury.">Axotomized bulbospinal neurons express c-Jun after cervical spinal cord injury.</a> Neuroreport. 16, 1535-1539 (2005).</li><li>Guenther, C. H., Windelborn, J. A., Tubon, T. C., Yin, J. C., Mitchell, G. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increased+atypical+PKC+expression+and+activity+in+the+phrenic+motor+nucleus+following+cervical+spinal+injury.">Increased atypical PKC expression and activity in the phrenic motor nucleus following cervical spinal injury.</a> Exp Neurol. 234, 513-520 (2012).</li><li>Mantilla, C. B., Gransee, H. M., Zhan, W. Z., Sieck, G. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motoneuron+BDNF/TrkB+signaling+enhances+functional+recovery+after+cervical+spinal+cord+injury.">Motoneuron BDNF/TrkB signaling enhances functional recovery after cervical spinal cord injury.</a> Exp Neurol. 247, 101-109 (2013).</li><li>Vinit, S., Darlot, F., Aoulaiche, H., Boulenguez, P., Kastner, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Distinct+expression+of+c-Jun+and+HSP27+in+axotomized+and+spared+bulbospinal+neurons+after+cervical+spinal+cord+injury.">Distinct expression of c-Jun and HSP27 in axotomized and spared bulbospinal neurons after cervical spinal cord injury.</a> J Mol Neurosci. 45, 119-133 (2011).</li><li>Windelborn, J. A., Mitchell, G. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glial+activation+in+the+spinal+ventral+horn+caudal+to+cervical+injury.">Glial activation in the spinal ventral horn caudal to cervical injury.</a> Respir Physiol Neurobiol. 180, 61-68 (2012).</li><li>Vinit, S., Stamegna, J. C., Boulenguez, P., Gauthier, P., Kastner, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Restorative+respiratory+pathways+after+partial+cervical+spinal+cord+injury:+role+of+ipsilateral+phrenic+afferents.">Restorative respiratory pathways after partial cervical spinal cord injury: role of ipsilateral phrenic afferents.</a> Eur J Neurosci. 25, 3551-3560 (2007).</li><li>Dougherty, B. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recovery+of+inspiratory+intercostal+muscle+activity+following+high+cervical+hemisection.">Recovery of inspiratory intercostal muscle activity following high cervical hemisection.</a> Respir Physiol Neurobiol. 183, 186-192 (2012).</li></ol>

Gli autori dichiarano di non avere interessi finanziari in competizione.

Difficoltà tecniche di rendere il Injury Modello C2 Il modello murino lesione C2 è un interessante strumento per lo studio delle vie respiratorie neuroplasticità post-lesionale. Tuttavia, i passi necessari per produrre un modello riproducibile e affidabile sono numerosi e ciascuno di essi potrebbero avere un impatto sui risultati dello studio. Ad esempio, durante il processo di intubazione, estrema cura deve essere presa in quanto il tubo orotracheale può produrre una in...

Trauma del midollo spinale è un pregiudizio comune osservata nella popolazione umana con incidenze drammatici, come la paralisi permanente. Tuttavia, la gravità del danno dipende dal livello e l&#39;entità del trauma iniziale. L&#39;insufficienza respiratoria è la causa principale della mortalità a seguito di lesione superiore cervicale del midollo spinale (SCI) 1. Attualmente, l&#39;unico trattamento terapeutico è quello di posizionare il paziente in ventilazione assistita. Da alcuni pazienti possono essere svezzati fuori l&#39;assistenza ventilatoria 2, a causa del recupero spontaneo che si verifica con ritardo post-lesionale, la necessità di sviluppare nuove terapie non invasive innovative è urgente 3. Avere un buon modello preclinico standardizzato per studiare l&#39;effetto di un SCI cervicale su insufficienza respiratoria e perciò studiare l&#39;applicazione di putativi strategie terapeutiche, è essenziale. In questo articolo tecnico, si descrive una specifica pre-clinica modello murino oinsufficienza respiratoria f indotta da una parziale SCI cervicale al livello C2. Questo modello è attualmente utilizzato da diversi laboratori di tutto il mondo (per le recensioni: 4-13). Tuttavia, lievi differenze nella procedura chirurgica possono essere osservati tra i diversi ricercatori per generare questo particolare cervicale lesione modello murino. L&#39;effetto di un C2 SCI sull&#39;uscita respiratorio è stato descritto nel 1895 da Porter 14. Un hemisection cervicale induce una deafferentazione dei motoneuroni frenico dalla loro unità centrale (nella rVRG nel tronco cerebrale, Figura 1A) sul lato omolaterale di lesioni, portando ad una attività del nervo frenico silenziosa e la successiva paralisi diaframma. Il lato controlaterale rimane intatto e permette all&#39;animale di sopravvivere. Diversamente differente SCI situato in un segmento vertebrale inferiore (ad esempio una lesione contusivi a livello C4 15), l&#39;integrità del nucleo motoneurone frenico su entrambi i lati è conservato. Dopo un CERVlesione C2 iCal, alcune attività spontanea può essere osservata sul lato ipsilaterale (frenico e diaframma) a causa di una attivazione di controlaterale sinapsi silenti che attraversavano la linea mediana della colonna vertebrale a livello segmentale C3-C6 (percorsi frenico incrociate, CPP, Figura 1B) . L&#39;attivazione del CPP, che è, per definizione, un hemisection C2 combinato con un phrenicotomy controlaterale che inducono un ipsilaterale parziale recupero del nervo frenico, può verificarsi da ore a settimane post-lesione 16-18. Il vero effetto benefico di questa via CPP sul recupero respiratorio è limitata 19 e ulteriori indagini e il trattamento deve essere sviluppata per migliorare la grandezza di restauro spontanea 3. Questo protocollo fornisce un potente tipo di modello murino pre-clinico per lo studio delle vie respiratorie plasticità post-lesionale a vari livelli (fisiologia respiratoria da motoneuroni pre e frenico, interneuroni, molecolare e cellular, locomozione dell&#39;arto anteriore per esempio), nonché un modello per verificare invasive e non invasive strategie terapeutiche mirate per migliorare la respirazione e il recupero motorio seguente lesione del midollo spinale cervicale C2 parziale.

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

Una lesione del midollo spinale cervicale induce paralisi permanente, e spesso porta a distress respiratorio. Ad oggi non terapeutica efficienti sono stati sviluppati per migliorare / migliorare l&#39;insufficienza respiratoria dopo un trauma ad alta cervicale del midollo spinale (SCI). Qui proponiamo un modello pre-clinico murino di alta SCI al 2 (C2) livello metameric cervicale per studiare diverse neuroplasticità respiratoria post-lesionale. La tecnica consiste di una lesione chirurgica parziale al livello C2, che indurrà una hemiparalysis del diaframma a causa di una deafferentazione dei motoneuroni frenico dai centri respiratori situati nel tronco cerebrale. Il lato controlaterale della lesione rimane intatto e permette il recupero degli animali. A differenza di altri siti di importanza comunitaria che influenzano la funzione locomotoria (a livello toracico e lombare), la funzione respiratoria non necessita di motivazione degli animali e la quantificazione del deficit / recupero può essere facilmente effettuata (diaframma e la registrazione del nervo frenicos, tutto ventilazione corpo). Questa pre-clinica modello C2 SCI è un modello pre-clinico potente, utile e affidabile per studiare vari eventi neuroplasticità delle vie respiratorie e non respiratorie a diversi livelli (molecolare alla fisiologia) e testare diverse putativi strategie terapeutiche che potrebbero migliorare la respirazione in pazienti SCI.

Entità della lesione Il successo e la riproducibilità di questo particolare modello sperimentale dipendono l&#39;esperienza di ciascun manipolatore / chirurgo. La successiva quantità di recupero respiratorio (attività del nervo frenico e del diaframma) a seguito di una lesione C2 è correlata con la restante ventrolateral risparmiato materia bianca 21. Poiché la lesione è &quot;fatto a mano&quot; e richiede una certa pratica del chirurgo, l&#39;entità di og...

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

L&#39;insufficienza respiratoria è la principale causa di morte in seguito ad una lesione del midollo spinale cervicale. Avere un modello animale preclinico riproducibile, quantificabili, e affidabile di insufficienza respiratoria indotta da una lesione cervicale parziale aiuterà a capire la successiva neuroplasticità respiratorie e non respiratorie e consentire strategie di sperimentazione di riparazione putative.

Un modello murino di cervicale lesioni del midollo spinale per studiare post-lesionale respiratoria neuroplasticità

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

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Research

JoVE Journal

Medicine

13.4K Views.  Université de Versailles Saint-Quentin-en-Yvelines.  L'insufficienza respiratoria è la principale causa di morte in seguito ad una lesione del midollo spinale cervicale. Avere un modello animale preclinico riproducibile, quantificabili, e affidabile di insufficienza respiratoria indotta da una lesione cervicale parziale aiuterà a capire la successiva neuroplasticità respiratorie e non respiratorie e consentire strategie di sperimentazione di riparazione putative. 

Video: 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.

Intraspinale trapianto di cellule cervicali targeting per corno ventrale nella Sclerosi Laterale Amiotrofica e le lesioni traumatiche del midollo spinale

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

Ricerca

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.

Acuta e Cronica test sensoriale tattile dopo lesione del midollo spinale in ratti

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.

Un modello contusivo unilaterale di lesioni del midollo spinale cervicale Uso della Impactor Infinite Horizon

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.

Controllata cervicale lesioni lacerazione nei topi

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.

Un modello Contusione di gravi lesioni del midollo spinale nei ratti

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.

Valutazione delle vie respiratorie di attivazione muscolare Utilizzando respiratoria Motor Control Assessment (RMCA) in soggetti con lesioni spinali croniche

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.

Controllata corticale modello Impact per trauma cranico

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.

Un sistema radio-telemetria per monitorare la funzionalità cardiovascolare in ratti con midollo spinale e Transection embrionali staminali neurali Innesti cellulari

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.

Utilizzo sinergico delle cellule neurali e precursori peptidi autoassemblanti in Sperimentale 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.

Calibrato forcipe Modello di Spinal Cord Injury 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 ...