LT ha ricevuto sovvenzioni da parte del Forschungsförderungsprogramm della Medicina dell&#39;Università di Göttingen. PL e MB sono stati sostenuti dal Centro di Ricerca per la DFG Fisiologia molecolare del cervello (CMPB), Göttingen. Gli autori ringraziano il Dott. Lars Tatenhorst per l&#39;assistenza e la videografia con Birgit Liebau aiuto con audio e video editing.

Gli animali sono stati acquistati da Jackson Laboratory (# 002.726) 1. Essi sono clinicamente segnato e sottoposto ad un test di funzionamento del motore (rotarod test) e della forza muscolare (appesa prova del filo). Tutti questi test e il successivo abbattimento degli animali al fine di preparare il midollo spinale sono stati eseguiti in accordo molto vicino alle linee guida locali per corretto svolgimento degli esperimenti sugli animali. 1. Punteggio clinico Oltre alla verifica del peso corporeo per i topi vengono esaminati i segni di deficit motorio con il seguente sistema di quattro punti di punteggio 2: 4 punti: normale (nessun segno di disfunzione motoria) 3 punti: tremori agli arti posteriori sono evidenti quando sospesa per la coda 2 punti: sono presenti anomalie dell&#39;andatura 1 punto: trascinamento di almeno un arto posteriore 0 punti: simmetricaparalisi, incapacità di destra stessa o perdita del 20% del peso corporeo massimo, in questo caso gli animali vengono sacrificati ed immediatamente l&#39;esperimento viene interrotto 2. Test della funzione motoria e della forza muscolare Appeso filo Questo test viene utilizzato per valutare la forza muscolare 3, 4. Tutti gli animali eseguire questo test almeno uno o due giorni dopo la prova rotarod. Ogni mouse è posizionato su una misura coperchio filo con intervalli di 0,8 cm e con cautela capovolto, 60 cm al di sopra un filo di paglia coperto fondo. Dopo l&#39;allenamento per tre volte consecutive di almeno 180 s la latenza a cadere è misurata. Ogni mouse è dato fino a tre tentativi di trattenere il coperchio capovolto per un massimo di 180 s e il più lungo periodo è registrata. Rotarod test <p class = &quot;jove_content&quot;&gt; L&#39;apparato rotarod (Ugo Basile, Comerio, Italia) è stato usato per misurare la coordinazione motoria, l&#39;equilibrio e la capacità di apprendimento motorio 3, 4. Una buona prestazione richiede un alto grado di coordinazione sensomotoria. La macchina deve essere collocato in un ambiente tranquillo e non disturba al fine di evitare stimoli di distrazione per la testati su animali. È costituita da un computer controllato motorizzata di rotazione del mandrino e cinque corsie per cinque topi. Cadute dei topi vengono rilevati automaticamente dalla pressione su una piastra di plastica sul fondo. Dopo l&#39;addestramento per tre volte consecutive di almeno 180 s ad una velocità costante di 15 rpm del tempo per il quale un animale può rimanere sulla canna rotante viene misurata. Ogni animale sottoposto a tre prove e la più lunga latenza senza cadere viene registrato. Il tempo di 180 s è scelto come cut-off time perché la maggior parte delle differenze significative nella coordinazione motoria vengono rilevati in questo lasso di tempo. 3. Preparazione Spinal Cord <ol class="lalpha"><li> Gli animali vengono uccisi da CO 2 insufflazione in conformità con le linee guida locali e sono immediatamente perfusi transcardiaca con soluzione PBS seguita da una soluzione di paraformaldeide al 4%. </li><li> Per preparare il midollo spinale del mouse sacrificato, l&#39;animale viene posto su un tavolo operatorio e le quattro arti sono fissati sul lato in modo da esporre il lato posteriore del mouse. </li><li> A breve lavaggio con una soluzione di etanolo al 70% pulisce il sito della dissezione e appiattisce il mantello. </li><li> Poi della cute con un bisturi affilato nella linea mediana. Al fine di facilitare il taglio della pelle viene tesa su entrambi i lati. Se i muscoli delle gambe deve essere pronto, la loro pelle deve inoltre essere inciso. </li><li> Dopo l&#39;incisione cutanea è completata, viene tirato da parte con una pinzetta per esporre la fascia sottostante superficiale del corpo. </li><li> La muscolatura del collo e il legamento nucale devono essere rimossi e sono carefully preparato. Fare attenzione a non incidere profondamente e di lesione del midollo spinale. I muscoli delle spalle possono anche essere rimosse per esporre meglio la colonna vertebrale. </li><li> Poi i muscoli paravertebrali vengono rimossi dalla intera colonna vertebrale. </li><li> Per aprire i laminectomie colonna vertebrale diversi devono essere eseguiti. Si dovrebbe iniziare dalla parte superiore craniale nel sito del atlanto-occipitale. </li><li> E &#39;più facile togliere la fissazione dei due arti superiori e tirare troppo il collo per essere meglio in grado di eseguire la laminectomia delle vertebre prima. Questi sono tirato via senza toccare il esposta midollo spinale cervicale. </li><li> Più vertebre vengono rimossi prima sezionare i archi vertebrali su entrambi i lati con forbici angolate e quindi tirando i processi dorsali. Rimanenti parti laterali delle vertebre dovrebbe essere rimosso per facilitare la successiva rimozione completa del midollo spinale. </li><li> Un punto di riferimento anatomico della spina lombarel è il cavo di intumescenza che è presente anche nel midollo spinale cervicale. </li><li> Dopo aver finito la laminectomia del midollo spinale intera, fare in modo che anche voi transetto tutte le radici ventrali e rilasciare il midollo spinale dalla dura madre delle meningi. </li><li> Poi il midollo spinale cervicale è tagliato cranialmente e si inizia a rimuovere il midollo spinale. </li><li> Infine, il midollo spinale è anche tagliato alla distale cauda equina, per essere completamente rilasciato. </li><li> In definitiva, il midollo spinale viene posto in una soluzione postfixating (ad esempio paraformaldeide 4%) per una notte e può essere ulteriormente lavorato. Di solito cryosection il midollo spinale per prepararlo per l&#39;analisi immunoistologico. </li></ol> 4. Risultati rappresentativi La tecnica di preparazione del midollo spinale rappresenta il focus di questo articolo video. Si tratta di un prerequisito essenziale per il sezionamento dei tessuti in seguito e, infine, per l&#39;analisi immunoistologico di spinal cavo sezioni. Come esempio di un risultato finale, uno workup immunoistochimica della regione corno anteriore del midollo spinale lombare topo di tipo selvatico (wt) e di un transgene SOD G93A (tg) topo è dimostrata. Neuroni motore può essere identificato con un primario anticorpo anti-ChAT e successiva marcatura fluorescente con un anticorpo secondario Cy3. Inoltre, una contro-colorazione nucleare con DAPI (4,6-Diamidino-2-fenilindolo) è stato eseguito (Figura 1). <img src="/files/ftp_upload/3936/3936fig1.jpg" alt="Figura 1" /> Figura 1. Microfotografie fluorescenti visualizzando il immunorivelazione dei motoneuroni con anticorpi anti-ChAT (rosso) e nuclei cellulari anti-macchia DAPI (blu) a livello lombare corno midollo spinale anteriore del mouse di un tipo selvatico (WT) (a sinistra) e di uno SOD G93A transgenico (tg) (a destra) del mouse all&#39;età di 130 giorni. Scale bar: 40 micron. Poiché l&#39;analisi immunoistochimica dii topi SOD G93A non è lo scopo primario di questo articolo si prega di consultare la pubblicazione originale in cui questi topi transgenici sono stati caratterizzati e quelli più recenti che studiano approcci terapeutici per la consultazione 1, 5, 6. Se gli effetti terapeutici è modulata al livello immunoistologico chiaramente definito algoritmi di valutazione quantitativa deve essere applicato supportato da un software stereological (si veda per esempio 7).

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Other Motor Neuron Disord. 6 (1), 55-62 (2005).</li><li>Zhong, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Activated+protein+C+therapy+slows+ALS&#45;like+disease+in+mice+by+transcriptionally+inhibiting+SOD1+in+motor+neurons+and+microglia+cells.">Activated protein C therapy slows ALS&#45;like disease in mice by transcriptionally inhibiting SOD1 in motor neurons and microglia cells.</a> J. Clin. Invest. 119 (11), 3437-3449 (2009).</li><li>Pitzer, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Granulocyte&#45;colony+stimulating+factor+improves+outcome+in+a+mouse+model+of+amyotrophic+lateral+sclerosis.">Granulocyte&#45;colony stimulating factor improves outcome in a mouse model of amyotrophic lateral sclerosis.</a> Brain. 131 (Pt. 12), 3335-3347 (2008).</li><li>Gowing, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ablation+of+proliferating+microglia+does+not+affect+motor+neuron+degeneration+in+amyotrophic+lateral+sclerosis+caused+by+mutant+superoxide+dismutase.">Ablation of proliferating microglia does not affect motor neuron degeneration in amyotrophic lateral sclerosis caused by mutant superoxide dismutase.</a> J. Neurosci. 28 (41), 10234-10244 (2008).</li><li>Scott, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=interpretation+of+studies+in+the+standard+murine+model+of+ALS.">interpretation of studies in the standard murine model of ALS.</a> Amyotroph Lateral Scler. 9 (1), 4-15 (2008).</li><li>Turner, B. J., Talbot, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transgenics,+toxicity+and+therapeutics+in+rodent+models+of+mutant+SOD1&#45;mediated+familial+ALS.">Transgenics, toxicity and therapeutics in rodent models of mutant SOD1&#45;mediated familial ALS.</a> Prog Neurobiol. 85 (1), 94-134 (2008).</li><li>Corse, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preclinical+testing+of+neuroprotective+neurotrophic+factors+in+a+model+of+chronic+motor+neuron+degeneration.">Preclinical testing of neuroprotective neurotrophic factors in a model of chronic motor neuron degeneration.</a> Neurobiol Dis. 6 (5), 335-346 (1999).</li><li>Knippenberg, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Significance+of+behavioural+tests+in+a+transgenic+mouse+model+of+amyotrophic+lateral+sclerosis+&#40;ALS&#41;.">Significance of behavioural tests in a transgenic mouse model of amyotrophic lateral sclerosis &#40;ALS&#41;.</a> Behav Brain Res. 213 (1), 82-87 (2010).</li><li>Burgess, R. W., Cox, G. A., Seburn, K. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuromuscular+disease+models+and+analysis.">Neuromuscular disease models and analysis.</a> Methods Mol. Biol. 602, 347-393 (2010).</li><li>Hayworth, C. R., Gonzalez-Lima, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pre-symptomatic+detection+of+chronic+motor+deficits+and+genotype+prediction+in+congenic+B6.SOD1(G93A)+ALS+mouse+model.">Pre-symptomatic detection of chronic motor deficits and genotype prediction in congenic B6.SOD1(G93A) ALS mouse model.</a> Neuroscience. 164 (3), 975-985 (2009).</li><li>Ludolph, A. 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Il (G93A) modello murino SOD1 genetico è un modello animale valido per studiare il corso della malattia del motoneurone perdita progressiva paragonabile umana sclerosi amiotrofica laterale 8. Una varietà di paradigmi di trattamenti diversi sono stati valutati in questo modello e rappresentano una base per la successiva prova in studi clinici umani 8-10. Per poter rilevare differenze significative in uno studio sperimentale di trattamento in questi topi, è di importanza eminente includere almeno ...

<table border="1"><tbody><tr><td></td><td> Fabbricante </td><td> Informazioni per l&#39;ordinazione </td></tr><tr><td> Test comportamentale </td><td></td><td></td></tr><tr><td> Rota-Rod per il Mice </td><td> Ugo Basile, Comerio, Italia </td><td> # 47600 </td></tr><tr><td> Hanging dispositivo a filo </td><td> Su misura </td><td></td></tr><tr><td colspan="3"></td></tr><tr><td> Preparazione Spinal Cord </td><td></td><td></td></tr><tr><td> Operation Table Spia di funzionamento Guanti di protezione </td><td></td><td></td></tr><tr><td> &quot;Iris&quot; Forbici, angolato a lato </td><td> Strumenti Scienza Belle, Heidelberg, Germania </td><td> 14063-09 </td></tr><tr><td> Cohan-Vännäs Forbici primavera, diritte </td><td> Strumenti Scienza Belle, Heidelberg, Germania </td><td> 15000-10 </td&gt; </tr><tr><td> Micro pinza </td><td> Hammacher, Solingen, Germania </td><td> HWC 111-10 </td></tr><tr><td> Scalpel &quot;präzisa plus&quot; </td><td> Dahlhausen, Köln, Germania </td><td> 11.000.00.510, figura 10 </td></tr></tbody></table>

clinical testing and spinal cord removal in a mouse model for amyotrophic lateral sclerosis (als)

La sclerosi laterale amiotrofica (SLA) è una malattia neurodegenerativa letale con conseguente progressiva degenerazione dei motoneuroni. Il picco di insorgenza è di circa 60 anni per la malattia sporadica e circa 50 anni per la malattia familiare. Grazie al suo andamento progressivo, il 50% dei pazienti muore entro 30 mesi dall&#39;inizio dei sintomi. Per valutare nuove opzioni di trattamento per questa malattia, modelli murini di SLA genetici sono stati generati sulla base umani mutazioni nel gene familiari SOD, come la (G93A) mutazione SOD1. Aspetti più importanti che devono essere valutati nel modello sono la sopravvivenza globale, decorso clinico e la funzione motoria. Qui, dimostriamo la valutazione clinica, mostrano la conduzione di due test del motore comportamentali e fornire sistemi di punteggio per tutti i parametri quantitativi. Poiché un&#39;analisi approfondita del modello murino ALS di solito richiede un esame immunoistochimica del midollo spinale, abbiamo dimostrato la sua preparazione in dettaglio l&#39;applicazione del farelaminectomia metodo rsal. Esemplari risultati istologici sono dimostrati. La completa applicazione dei metodi di esame descritti negli studi sul modello murino di SLA consentirà al ricercatore di verificare in modo affidabile per future opzioni terapeutiche che possono fornire una base per le sperimentazioni cliniche umane successive.

Watch this Scientific Journal Video about Clinical Testing and Spinal Cord Removal in a Mouse Model for Amyotrophic Lateral Sclerosis ALS at JoVE.com

Clinical Testing and Spinal Cord Removal in a Mouse Model for Amyotrophic Lateral Sclerosis ALS

Un modello murino per la sclerosi laterale amiotrofica (SLA) è una valutazione clinica e comportamentale. Come prerequisito per un&#39;analisi accompagnamento immunoistologico la preparazione del midollo spinale è raffigurato in dettaglio.

Test clinici e la rimozione del midollo spinale in un modello murino di sclerosi laterale amiotrofica (SLA)

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder resulting in progressive degeneration of motoneurons. Peak of onset is around 60 ...

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Research

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Medicine

Clinical Testing and Spinal Cord Removal in a Mouse Model for Amyotrophic Lateral Sclerosis (ALS)

27.9K Views.  University Medicine Göttingen. Un modello murino per la sclerosi laterale amiotrofica (SLA) è una valutazione clinica e comportamentale. Come prerequisito per un'analisi accompagnamento immunoistologico la preparazione del midollo spinale è raffigurato in dettaglio.

Video: Clinical Testing and Spinal Cord Removal in a Mouse Model for Amyotrophic Lateral Sclerosis ALS

A Protocol for Comprehensive Assessment of Bulbar Dysfunction in Amyotrophic Lateral Sclerosis (ALS)

Improved methods for assessing bulbar impairment are necessary for expediting diagnosis of bulbar dysfunction in ALS, for predicting disease progression across speech subsystems, and for addressing the critical need for sensitive outcome measures for ongoing experimental treatment trials. To address this need, we are obtaining longitudinal profiles of bulbar impairment in 100 individuals based on a comprehensive instrumentation-based assessment that yield objective measures. Using instrumental approaches to quantify speech-related behaviors is very important in a field that has primarily relied on subjective, auditory-perceptual forms of speech assessment1. Our assessment protocol measures performance across all of the speech subsystems, which include respiratory, phonatory (laryngeal), resonatory (velopharyngeal), and articulatory. The articulatory subsystem is divided into the facial components (jaw and lip), and the tongue. Prior research has suggested that each speech subsystem responds differently to neurological diseases such as ALS. The current protocol is designed to test the performance of each speech subsystem as independently from other subsystems as possible. The speech subsystems are evaluated in the context of more global changes to speech performance. These speech system level variables include speaking rate and intelligibility of speech.
The protocol requires specialized instrumentation, and commercial and custom software. The respiratory, phonatory, and resonatory subsystems are evaluated using pressure-flow (aerodynamic) and acoustic methods. The articulatory subsystem is assessed using 3D motion tracking techniques. The objective measures that are used to quantify bulbar impairment have been well established in the speech literature and show sensitivity to changes in bulbar function with disease progression. The result of the assessment is a comprehensive, across-subsystem performance profile for each participant. The profile, when compared to the same measures obtained from healthy controls, is used for diagnostic purposes. Currently, we are testing the sensitivity and specificity of these measures for diagnosis of ALS and for predicting the rate of disease progression. In the long term, the more refined endophenotype of bulbar ALS derived from this work is expected to strengthen future efforts to identify the genetic loci of ALS and improve diagnostic and treatment specificity of the disease as a whole. The objective assessment that is demonstrated in this video may be used to assess a broad range of speech motor impairments, including those related to stroke, traumatic brain injury, multiple sclerosis, and Parkinson disease.

A Protocol for Comprehensive Assessment of Bulbar Dysfunction in Amyotrophic Lateral Sclerosis ALS

Objective assessments of the physiological mechanisms that support speech are needed to monitor disease onset and progression in persons with ALS and to quantify treatment effects in clinical trials. In this video, we present a comprehensive, instrumentation-based protocol for quantifying speech motor performance in clinical populations.

Un protocollo per la valutazione completa della disfunzione bulbare nella Sclerosi Laterale Amiotrofica (SLA)

Improved methods for assessing bulbar impairment are necessary for expediting diagnosis of bulbar dysfunction in ALS, for predicting disease progression ...

Ricerca

Medicina

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) ...

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 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 ...

Utility of Dissociated Intrinsic Hand Muscle Atrophy in the Diagnosis of Amyotrophic Lateral Sclerosis

The split hand phenomenon refers to predominant wasting of thenar muscles and is an early and specific feature of amyotrophic lateral sclerosis (ALS). A novel split hand index (SI) was developed to quantify the split hand phenomenon, and its diagnostic utility was assessed in ALS patients. The split hand index was derived by dividing the product of the compound muscle action potential (CMAP) amplitude recorded over the abductor pollicis brevis and first dorsal interosseous muscles by the CMAP amplitude recorded over the abductor digiti minimi muscle. In order to assess the diagnostic utility of the split hand index, ALS patients were prospectively assessed and their results were compared to neuromuscular disorder patients. The split hand index was significantly reduced in ALS when compared to neuromuscular disorder patients (P&#60;0.0001). Limb-onset ALS patients exhibited the greatest reduction in the split&#160;hand index, and a value of 5.2 or less reliably differentiated ALS from other neuromuscular disorders. Consequently, the split&#160;hand index appears to be a novel diagnostic biomarker for ALS, perhaps facilitating an earlier diagnosis.

Dissociated atrophy of intrinsic hand muscles, termed the split hand, appears to be a specific feature of amyotrophic lateral sclerosis (ALS).&#160;Consequently, a novel neurodiagnostic test, termed the split hand&#160;index, was developed to quantify the clinical phenomenon of the split hand, which differentiated ALS from mimic disorders.

Utilità di dissociata intrinseca mano Atrofia muscolare nella diagnosi di Sclerosi Laterale Amiotrofica

The split hand phenomenon refers to predominant wasting of thenar muscles and is an early and specific feature of amyotrophic lateral sclerosis (ALS). A ...

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 ...

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 ...

A Quick Phenotypic Neurological Scoring System for Evaluating Disease Progression in the SOD1-G93A Mouse Model of ALS

The SOD1-G93A transgenic mouse is the most widely used animal model of amyotrophic lateral sclerosis (ALS). At ALS TDI we developed a phenotypic screening protocol, demonstrated in video herein, which reliably assesses the neuromuscular function of SOD1-G93A mice in a quick manner. This protocol encompasses a simple neurological scoring system (NeuroScore) designed to assess hindlimb function. NeuroScore is focused on hindlimb function because hindlimb deficits are the earliest reported neurological sign of disease in SOD1-G93A mice. The protocol developed by ALS TDI provides an unbiased assessment of onset of paresis (slight or partial paralysis), progression and severity of paralysis and it is sensitive enough to identify drug-induced changes in disease progression. In this report, the combination of a detailed manuscript with video minimizes scoring ambiguities and inter-experimenter variability thus allowing for the protocol to be adopted by other laboratories and enabling comparisons between studies taking place at different settings. We believe that this video protocol can serve as an excellent training tool for present and future ALS researchers.

This video protocol describes a sensitive, reliable, and quick method for evaluating the neuromuscular deficits in a transgenic mouse model of amyotrophic lateral sclerosis.

Un Sistema di punteggio rapida fenotipica neurologica per la valutazione di progressione della malattia nel SOD1-G93A modello murino della SLA

The SOD1-G93A transgenic mouse is the most widely used animal model of amyotrophic lateral sclerosis (ALS). At ALS TDI we developed a phenotypic screening ...

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.

Un neonatale mouse Spinal Cord Injury compressione Modello

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