LT ha recibido apoyo económico del Forschungsförderungsprogramm de la Medicina de la Universidad de Gotinga. PL y MB fueron apoyados por el Centro de Investigación DFG para la Fisiología Molecular del Cerebro (CMPB), Göttingen. Los autores agradecen al Dr. Lars Tatenhorst para obtener ayuda con la videografía y Liebau Birgit ayuda para edición de audio y vídeo.

Los animales fueron adquiridos de Jackson Laboratory (# 002726) 1. Se anotó clínicamente y se sometió a una prueba de función motora (rotarod prueba) y de la fuerza muscular (colgando prueba de hilo). Todas estas pruebas y el asesinato posterior de los animales a fin de preparar la médula espinal se han realizado de acuerdo muy cerca de las directrices locales para la correcta realización de los experimentos con animales. 1. Puntuación clínica Aparte de la evaluación para los ratones de peso corporal se examinan en busca de signos de déficit motor con el siguiente sistema de puntuación de 4, punto 2: 4 puntos: normal (sin signos de disfunción motora) 3 puntos: los temblores de las extremidades traseras son evidentes cuando se suspendió por la cola 2 puntos: alteraciones de la marcha están presentes 1 punto: arrastrando de por lo menos una extremidad posterior 0 puntos: simétricaparálisis, incapacidad para derecho propio o pérdida de 20% del peso corporal máximo, en este caso los animales son sacrificados inmediatamente y el experimento se termina 2. Las pruebas de la función motora y la fuerza muscular Suspensión de la cuerda Esta prueba se utiliza para evaluar la fuerza muscular 3, 4. Todos los animales de realizar esta prueba por lo menos uno o dos días después de la prueba rotarod. Cada ratón se sitúa sobre una tapa de alambre a la medida, con intervalos de 0,8 cm y con cautela al revés, 60 cm por encima de una cubierta de paja inferior. Después de entrenar durante tres veces consecutivas de al menos 180 s la latencia se mide a caer. Cada ratón se da hasta tres intentos de aferrarse a la tapa invertida por un máximo de 180 s, y el período más largo se registra. Rotarod prueba 3, 4. Un buen desempeño requiere de un alto grado de coordinación sensoriomotora. La máquina debe ser colocado en un ambiente tranquilo y no molesto a evitar los estímulos de distracción para el animal a prueba. Se compone de un equipo controlado accionado por motor del husillo giratorio y cinco carriles durante cinco ratones. Caídas de los ratones se detectan automáticamente por la presión sobre una placa de plástico en la parte inferior. Después de la formación de tres veces consecutivas de al menos 180 s a una velocidad constante de 15 rpm, el tiempo para que un animal puede permanecer en la varilla giratoria se mide. Cada animal se somete a tres pruebas y el más largo de latencia, sin caer se registra. El tiempo de 180 s es elegido como corte de tiempo porque la mayoría de las diferencias significativas en la coordinación motora se detectan en este período de tiempo. 3. Preparación de la médula espinal <ol class="lalpha"><li> Los animales se mataron por insuflación de CO 2 de acuerdo con las directrices locales y son inmediatamente perfundidos transcardialmente con solución de PBS seguido por una solución de paraformaldehído al 4%. </li><li> A fin de preparar la médula espinal del ratón sacrificado, el animal se coloca sobre una mesa de operaciones y los cuatro miembros están fijos en el lado con el fin de exponer el lado posterior del ratón. </li><li> Un corto de lavado con una solución de etanol al 70% limpia el sitio de la disección y se aplana la capa de pelo. </li><li> A continuación, la piel se realiza una incisión con un bisturí afilado en la línea media. Con el fin de facilitar el corte de la piel se estira a ambos lados. Si los músculos de las piernas se prepararán, su piel también ha de realizarse una incisión. </li><li> Después de la incisión de la piel se ha completado, se llevó a un lado con un par de pinzas para exponer la fascia subyacente superficial del cuerpo. </li><li> La musculatura del cuello y el ligamento nucal tiene que ser eliminado y son carefully preparado. Tenga cuidado de no incidir en profundidad y la lesión de la médula espinal. Los músculos de los hombros también se puede quitar con el fin de exponer mejor la columna vertebral. </li><li> Entonces los músculos paravertebrales se eliminan de toda la columna vertebral. </li><li> Con el fin de abrir los laminectomías columna vertebral varios tienen que ser realizadas. Uno debe comenzar desde la parte superior de cráneo en el sitio de la articulación atlanto-occipital. </li><li> Es más fácil para eliminar la fijación de la parte superior de dos miembros y estirar el cuello para estar en mejores condiciones para realizar la laminectomía de la primera vértebra. Estos se alejó, sin tocar la médula espinal cervical expuesta. </li><li> Más vértebras se separan por primera transecting los arcos vertebrales en ambos lados con unas tijeras en ángulo y luego tirando de los procesos dorsales. El resto de las partes laterales de las vértebras debe ser eliminado para facilitar el posterior eliminación completa de la médula espinal. </li><li> Un punto de referencia anatómica de la espina lumbarcable l es la intumescencia que también está presente en la médula espinal cervical. </li><li> Una vez terminada la laminectomía de la médula espinal completa, asegúrese de que usted también seccionar todas las raíces ventrales y liberar la médula espinal de la duramadre de las meninges. </li><li> A continuación, la médula espinal cervical se corta cranealmente y empezar a eliminar la médula espinal. </li><li> Finalmente la médula espinal también se corta en la parte distal cauda equina, para ser completamente liberado. </li><li> En última instancia, la médula espinal se coloca en una solución postfixating (por ejemplo paraformaldehído al 4%) durante la noche y se puede procesar más. Por lo general, criosección la médula espinal para prepararlo para el análisis inmunohistoquímico. </li></ol> 4. Los resultados representativos La técnica de preparación de la médula espinal representa el foco de este artículo de vídeo. Es un requisito previo esencial para la sección de tejido más tarde y en última instancia, para el análisis inmunohistoquímico de la espinal secciones de la médula. Como un ejemplo de un resultado final, un estudio diagnóstico inmunohistoquímico de la región asta anterior de la médula espinal lumbar ratón de un tipo salvaje (wt) y de un SOD G93A transgén (TG) ratón se demuestra. Las neuronas motoras pueden ser identificados con un anticuerpo primario anti-CHAT anticuerpos y su posterior marcado fluorescente con una secundaria Cy3 anticuerpo. Además, un contador nuclear-mancha con DAPI (4,6-diamino-2-fenilindol) se ha realizado (Figura 1). <img src="/files/ftp_upload/3936/3936fig1.jpg" alt="Figura 1" /> Figura 1. Fotomicrografías fluorescentes visualizar la inmunodetección de las motoneuronas con anti-CHAT anticuerpos (rojo) y núcleos celulares contra-tinción con DAPI (azul) en el ratón de la médula espinal lumbar de cuerno anterior de un tipo salvaje (wt) (a la izquierda) y de un SOD G93A transgénico (TG) (a la derecha) del ratón a la edad de 130 días. Barra de escala: 40 m. A medida que el análisis inmunohistoquímico delos ratones SOD G93A no es el ámbito principal de este artículo, consulte la publicación original en el que estos ratones transgénicos han sido caracterizados y otras más recientes que estudian los enfoques terapéuticos para la referencia adicional de 1, 5, 6. Si los efectos terapéuticos se diferencian en el nivel inmunohistoquímico claramente definido algoritmos cuantitativos de evaluación debe ser aplicado con el apoyo de un software estereológicos (por ejemplo, ver 7).

<ol>
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N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+phenotyping+strategies+for+mutant+mice.">Behavioral phenotyping strategies for mutant mice.</a> Neuron. 57 (6), 809-818 (2008).</li><li>Miana-Mena, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimal+methods+to+characterize+the+G93A+mouse+model+of+ALS.">Optimal methods to characterize the G93A mouse model of ALS.</a> Amyotroph. Lateral Scler. 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. 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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. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Guidelines+for+preclinical+animal+research+in+ALS&#47;MND:+A+consensus+meeting.">Guidelines for preclinical animal research in ALS&#47;MND: A consensus meeting.</a> Amyotroph Lateral Scler. 11 (1-2), 38-45 (2010).</li><li>Boillee, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Onset+and+progression+in+inherited+ALS+determined+by+motor+neurons+and+microglia.">Onset and progression in inherited ALS determined by motor neurons and microglia.</a> Science. 5778 (3), 1389-1392 (2006).</li></ol>

No hay conflictos de interés declarado.

La SOD1 (G93A) modelo de ratón genético es un modelo animal útil para estudiar el curso de la enfermedad de la pérdida de las motoneuronas progresiva comparable a la humana esclerosis lateral amiotrófica 8. Una variedad de diferentes modelos de tratamientos se han evaluado en este modelo y representan una base para después probar en humanos 8-10 estudios clínicos. Con el fin de ser capaz de detectar diferencias significativas en un estudio de tratamiento experimental en estos ratones, es de ...

<table border="1"><tbody><tr><td></td><td> Fabricante </td><td> Información para hacer pedidos </td></tr><tr><td> Las pruebas de comportamiento </td><td></td><td></td></tr><tr><td> Rota-Rod de los ratones </td><td> Ugo Basile, Comerio, Italia </td><td> # 47600 </td></tr><tr><td> Colgar el dispositivo de alambre </td><td> Hechas a medida </td><td></td></tr><tr><td colspan="3"></td></tr><tr><td> Preparación de la médula espinal </td><td></td><td></td></tr><tr><td> Operación de la tabla Funcionamiento de la lámpara Guantes de protección </td><td></td><td></td></tr><tr><td> &quot;Iris&quot; Tijeras, en ángulo a otro </td><td> Herramientas de Bellas Ciencia, Heidelberg, Alemania </td><td> 14063-09 </td></tr><tr><td> Cohan-Vannas Tijeras rectas de primavera, </td><td> Herramientas de Bellas Ciencia, Heidelberg, Alemania </td><td> 15000-10 </td&gt; </tr><tr><td> Micro pinzas </td><td> Hammacher, Solingen, Alemania </td><td> HWC 111-10 </td></tr><tr><td> Bisturí &quot;präzisa más&quot; </td><td> Dahlhausen, Colonia, Alemania </td><td> 11.000.00.510, la figura 10 </td></tr></tbody></table>

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

La esclerosis lateral amiotrófica (ELA) es un trastorno neurodegenerativo fatal que resulta en la degeneración progresiva de neuronas motoras. Pico de aparición es de alrededor de 60 años de la enfermedad esporádica y alrededor de 50 años para la enfermedad familiar. Debido a su curso progresivo, el 50% de los pacientes mueren dentro de los 30 meses del inicio de los síntomas. Con el fin de evaluar nuevas opciones de tratamiento para esta enfermedad, los modelos genéticos de ratón de la ALS se han generado sobre la base de las mutaciones humanas familiares en el gen SOD, tales como la SOD1 (G93A) mutación. Aspectos más importantes que tienen que ser evaluadas en el modelo son la supervivencia global, la evolución clínica y la función motora. En este caso, se demuestra la evaluación clínica, muestran la realización de dos pruebas de motor de comportamiento y proporcionar sistemas cuantitativos de calificación para todos los parámetros. Debido a un análisis en profundidad del modelo de ratón de ALS por lo general requiere un examen inmunohistoquímico de la médula espinal, que demuestran su preparación en detalle la aplicación de la vermétodo de laminectomía rsal. Los ejemplos de los hallazgos histológicos se ha demostrado. La aplicación completa de los métodos de examen que aparecen en los estudios sobre el modelo de ratón de la ELA permitirá a los investigadores para probar de forma fiable las futuras opciones terapéuticas que pueden servir de base para los ensayos clínicos más humanos.

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 modelo de ratón para la esclerosis lateral amiotrófica (ELA) es un examen clínico y de comportamiento. Como un requisito previo para un análisis inmunohistoquímico acompaña la preparación de la médula espinal se representa en detalle.

Los ensayos clínicos y de eliminación de la médula espinal en un modelo murino de esclerosis lateral amiotrófica (ELA)

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

clinical-testing-spinal-cord-removal-mouse-model-for-amyotrophic

Research

JoVE Journal

Medicine

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

27.9K vistas.  University Medicine Göttingen. Un modelo de ratón para la esclerosis lateral amiotrófica (ELA) es un examen clínico y de comportamiento. Como un requisito previo para un análisis inmunohistoquímico acompaña la preparación de la médula espinal se representa en detalle.

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 Protocolo de Evaluación Integral de la disfunción bulbar de la Esclerosis Lateral Amiotrófica (ALS)

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

Investigación

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.

Intraespinal trasplante de células de Orientación asta ventral del cuello uterino en la esclerosis lateral amiotrófica y la lesión de la médula espinal

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.

Aguda y crónica Prueba sensorial táctil después de la lesión de la médula espinal en ratas

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 modelo Contusión de la lesión de la médula espinal severa en ratas

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.

Utilidad de la disociada intrínseca Mano Atrofia muscular en el diagnóstico de la esclerosis lateral amiotrófica

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.

Controlado Modelo de Impacto cortical de la lesión 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.

Un sistema de radio-telemetría para monitorizar la función cardiovascular en ratas con la médula espinal y transección Embryonic Stem Neural injertos de células

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.

Calibrado Pinzas Modelo de Lesión Medular Compresión

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 de puntuación rápida fenotípica Neurológica para la Evaluación de progresión de la enfermedad en el SOD1-G93A modelo de ratón de la ELA

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 ratón neonatal lesión de médula espinal de compresión Modelo

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