Ich möchte danken: allen Mitgliedern der Lepore, Maragakis und Rothstein Labors für hilfreiche Diskussionen, die gelähmten Veteranen von Amerika und der Craig H. Neilsen Foundation für finanzielle Unterstützung.

<ol>
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Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motor+neuron+degeneration+in+mice+that+express+a+human+Cu,Zn+superoxide+dismutase+mutation.">Motor neuron degeneration in mice that express a human Cu,Zn superoxide dismutase mutation.</a> Science. 264, 1772-1775 (1994).</li><li>Howland, D. S., Liu, J., She, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Focal+loss+of+the+glutamate+transporter+EAAT2+in+a+transgenic+rat+model+of+SOD1+mutant-mediated+amyotrophic+lateral+sclerosis+(ALS).">Focal loss of the glutamate transporter EAAT2 in a transgenic rat model of SOD1 mutant-mediated amyotrophic lateral sclerosis (ALS).</a> Proc Natl Acad Sci. 99, 1604-1604 (2002).</li><li>Nagai, M., Aoki, M., Miyoshi, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rats+expressing+human+cytosolic+copper-zinc+superoxide+dismutase+transgenes+with+amyotrophic+lateral+sclerosis:+associated+mutations+develop+motor+neuron+disease.">Rats expressing human cytosolic copper-zinc superoxide dismutase transgenes with amyotrophic lateral sclerosis: associated mutations develop motor neuron disease.</a> J. Neurosci. 21, 9246-9246 (2001).</li><li>McDonald, W., Becker, D. <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:+promising+interventions+and+realistic+goals.">Spinal cord injury: promising interventions and realistic goals.</a> Am. J. Phys. Med. Rehabi. I82, S38-S38 (2003).</li><li>Sandrow-Feinberg, H. R., Zhukareva, V., Santi, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=PEGylated+interferon-beta+modulates+the+acute+inflammatory+response+and+recovery+when+combined+with+forced+exercise+following+cervical+spinal+contusion+injury.">PEGylated interferon-beta modulates the acute inflammatory response and recovery when combined with forced exercise following cervical spinal contusion injury.</a> Experimental neurology. 223, 439-451 (2010).</li><li>Gage, F. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mammalian+neural+stem+cells.">Mammalian neural stem cells.</a> Science. 287, 1433-1433 (2000).</li><li>Lane, M. A., Lee, K. Z., 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=Spinal+circuitry+and+respiratory+recovery+following+spinal+cord+injury.">Spinal circuitry and respiratory recovery following spinal cord injury.</a> Respiratory physiology &#38; neurobiology. 169, 123-123 (2009).</li><li>Lepore, A. C., Rauck, B., Dejea, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Focal+transplantation-based+astrocyte+replacement+is+neuroprotective+in+a+model+of+motor+neuron+disease.">Focal transplantation-based astrocyte replacement is neuroprotective in a model of motor neuron disease.</a> Nature. 11, 1294-1294 (2008).</li><li>Rao, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multipotent+and+Restricted+Precursors+in+the+Central+Nervous+System.">Multipotent and Restricted Precursors in the Central Nervous System.</a> Anat Rec. 257, 137-137 (1999).</li><li>Rao, M. S., Mayer-Proschel, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glial-+restricted+precursors+are+derived+from+multipotent+neuroepithelial+stem+cells.">Glial- restricted precursors are derived from multipotent neuroepithelial stem cells.</a> Dev Biol. 188, 48-48 (1997).</li><li>Suzuki, M., Tork, C., Shelley, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sexual+dimorphism+in+disease+onset+and+progression+of+a+rat+model+of+ALS.+Sexual+dimorphism+in+disease+onset+and+progression+of+a+rat+model+of+ALS.">Sexual dimorphism in disease onset and progression of a rat model of ALS. Sexual dimorphism in disease onset and progression of a rat model of ALS.</a> Amyotroph Lateral Scler. 8, 20-20 (2007).</li><li>Lepore, A. C., Haenggeli, C., Gasmi, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intraparenchymal+spinal+cord+delivery+of+adeno-associated+virus+IGF-1+is+protective+in+the+SOD1G93A+model+of+ALS.">Intraparenchymal spinal cord delivery of adeno-associated virus IGF-1 is protective in the SOD1G93A model of ALS.</a> Brain research. 1185, 256-256 (2007).</li><li>Veldink, J. H., Bar, P. R., Joosten, E. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sexual+differences+in+onset+of+disease+and+response+to+exercise+in+a+transgenic+model+of+ALS.">Sexual differences in onset of disease and response to exercise in a transgenic model of ALS.</a> Neuromuscul Disord. 13, 737-737 (2003).</li><li>Shumsky, J. S., Lepore, 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=Transplantation+of+Neuronal+and+Glial+Restricted+Precursors+into+Contused+Spinal+Cord+Improves+Bladder+and+Motor+Functions,+Decreases+Thermal+Hypersensitivity,+and+Modifies+Intraspinal+Circuitry.">Transplantation of Neuronal and Glial Restricted Precursors into Contused Spinal Cord Improves Bladder and Motor Functions, Decreases Thermal Hypersensitivity, and Modifies Intraspinal Circuitry.</a> J. Neurosci. 25, 9624-9624 (2005).</li></ol>

Keine Interessenskonflikte erklärt.

Für Studien mit SOD1 G93A Mäusen und Ratten, Alter und Geschlecht entsprechen den Tieren innerhalb einer Gruppe und verteilen Tiere innerhalb der gleichen Wurf zu verschiedenen Gruppen. Es ist besser, alle Tiere aus dem gleichen Geschlecht für ALS und SCI-Modellen verwenden, da Krankheitsprozesse zwischen Männchen und Weibchen unterscheiden können, aber es kann auch nützlich sein, um genügend Tiere aus beiden Geschlechtern, um mögliche geschlechtsspezifische Wirkungen zu erkennen sind, wie Dieses Phä...

<table border="1"><tbody><tr><td> Name des Reagenz </td><td> Firma </td><td> Katalog-Nummer </td></tr><tr><td> HBSS </td><td> Gibco </td><td> 14170 </td></tr><tr><td> 0,05% Trypsin </td><td> Gibco </td><td> 25300 </td></tr><tr><td> Sojabohnen-Trypsin-Inhibitor (optional) </td><td> Sigma </td><td> T-6522 </td></tr><tr><td> Acepromazine Maleat (0,7 mg / kg) </td><td> Fermenta Animal Health </td><td></td></tr><tr><td> Ketamin (95 mg / kg) </td><td> Fort Dodge Animal Health </td><td></td></tr><tr><td> Xylazin (10 mg / kg) </td><td> Bayer </td><td></td></tr><tr><td> Nr. 11 Feather Skalpell </td><td> Electron Microscopy Sciences </td><td> 72044-11 </td></tr><tr><td> Wattestäbchen (6 inch) </td><td> Fischer </td><td> 23-400-101 </td></tr><tr><td> Rat-Hakenpinzette </td><td> Feine Science Tools </td><td> Rat: 11023-15; Maus: 11042-08 </td></tr><tr><td> Mittelständische Frühjahr Schere </td><td> Feine Science Tools </td><td> 15012-12 </td></tr><tr><td> Mini-Frühlingsrollen Schere </td><td> Feine Science Tools </td><td> 15000-10 </td></tr><tr><td> Rongeur </td><td> Feine Science Tools </td><td> Rat: 16121-14; Maus: 16221-14 </td></tr><tr><td> Microknife </td><td> Feine Science Tools </td><td> 10056-12 </td></tr><tr><td> Nadelhalter </td><td> Feine Science Tools </td><td> 12502-14 </td></tr><tr><td> Suture: 4-0 </td><td> Vicryl </td><td> S-183 </td></tr><tr><td> Staples: 9 mm </td><td> Autoclip </td><td> 427631 </td></tr><tr><td> Hefter: 9 mm (Reflex # 203-1000) </td><td> World Precision Instruments </td><td> 5000344 </td></tr><tr><td> Warm-Wasserpumpe (T / Pump) </td><td> Gaymar </td><td> P / N 07999-000 </td></tr><tr><td> Cyclosporin A: 250,0 mg/5.0 mL Ampullen </td><td> Novartis / Sandimmun </td><td> NDC 0078-0109-01 </td></tr><tr><td> FK-506 </td><td> LC Laboratories </td><td> F-4900 </td></tr><tr><td> Rapamycin </td><td> LC Laboratories </td><td> R-5000 </td></tr><tr><td> Injector </td><td> World Precision Instruments </td><td> UMP2 </td></tr><tr><td> Micro 4 Mikroliterspritze Pump Controller </td><td> World Precision Instruments </td><td> UMC4 </td></tr><tr><td> Mikromanipulator </td><td> World Precision Instruments </td><td> Kite-R </td></tr><tr><td> 10,0 ul Hamilton-Spritze </td><td> Hamilton </td><td> 80030 </td></tr><tr><td> Hamilton Nadeln: 33-gauge, 45 ° abgeschrägt, 1 Zoll </td><td> Hamilton </td><td> 7803-05 </td></tr><tr><td> Glass 20,0 ul Mikrokapillare Pipetten (optional) </td><td> Kimble </td><td> 71900-20 </td></tr></tbody></table>

intraspinal cell transplantation for targeting cervical ventral horn in amyotrophic lateral sclerosis and traumatic spinal cord injury

Respiratory Kompromiss aufgrund phrenicus Motoneuronen Verlust ist eine schwächende Folge ein großer Teil der Menschen traumatische Rückenmarksverletzungen (SCI) Fälle 1 und ist der ultimative Todesursache bei Patienten mit motorischen Neurons Erkrankung, amyotrophe seitlichen Sklerose (ALS) 2. ALS ist eine verheerende neurologische Erkrankung, die durch relativ schnelle Degeneration der oberen und unteren Motoneuronen gekennzeichnet ist. Patienten letztlich erliegen der Krankheit im Durchschnitt 2-5 Jahre nach der Diagnose aufgrund der Atemlähmung durch den Verlust von phrenicus Motoneuronen innnervation der Membran 3. Die überwiegende Mehrheit der Fälle sind sporadisch, während 10% der familiären Form sind. Etwa zwanzig Prozent der familiären Fälle sind auf verschiedene Punktmutationen in der Cu / Zn-Superoxiddismutase 1 (SOD1)-Gen auf Chromosom 21 4 verbunden. Transgene Mäuse und Ratten 4,5 6 Mitführen mutierten SOD1 menschlichen Gene (G93A, G37R, G86R, G85R) generiert wurden, und trotz der Existenz der anderen Tiermodellen der Motoneuronen Verlust, sind derzeit die am meisten verwendeten Modelle der Krankheit . Rückenmarksverletzung (SCI) ist eine heterogene Gruppe von Bedingungen, die aus physischen Trauma des Rückenmarks, mit dem funktionellen Ergebnis je nach der Art, Lage und Schwere der Verletzung 7. Dennoch beeinflussen etwa die Hälfte der menschlichen SCI Fällen Gebärmutterhalskrebs Regionen, die sich in lähmenden respiratorischen Dysfunktion aufgrund phrenicus Motoneuronen Verlust und Verletzung absteigend bulbospinal Atemwege Axone 1. Eine Reihe von Tiermodellen für SCI entwickelt worden, mit dem am häufigsten verwendeten und klinisch relevanter als die Prellung 8. Die Transplantation von verschiedenen Klassen von neuronalen Vorläuferzellen (NPCs) ist eine vielversprechende therapeutische Strategie für die Behandlung von traumatischen ZNS-Verletzungen und Neurodegeneration, einschließlich ALS und Rückenmarksverletzungen, wegen der Fähigkeit, verloren gegangene oder dysfunktionalen CNS Zelltypen, bieten Neuroprotektion zu ersetzen, und liefern Gen Faktoren von Interesse 9. Tiermodelle von ALS und SCI modellieren können viele klinisch-relevanten Aspekte dieser Krankheiten, einschließlich phrenicus Motoneuronen Verlust und damit der Atemwege Kompromiss 10,11. Um die Wirksamkeit von NPC-Strategien auf die Atemfunktion in diesen Tiermodellen der ALS und Rückenmarksverletzungen zu bewerten, müssen zelluläre Eingriffe speziell in Regionen mit therapeutisch relevanten Ziele wie phrenicus Motoneuronen gerichtet werden. Wir bieten Ihnen ein detailliertes Protokoll für Multi-Segment, intraspinale Transplantation von NPCs in den zervikalen Rückenmarks ventralen grauen Substanz von neurodegenerativen Modellen wie SOD1 G93A Mäusen und Ratten, sowie Rückenmark verletzt Ratten und Mäusen 11.

Watch this Scientific Journal Video about Intraspinal Cell Transplantation for Targeting Cervical Ventral Horn in Amyotrophic Lateral Sclerosis and Traumatic Spinal Cord Injury at JoVE.com

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

Neuronale Vorläuferzellen der Transplantation ist eine vielversprechende Strategie für den Schutz und / oder Wiederbeschaffung verlorener / dysfunktional Gebärmutterhals phrenicus Motoneuronen im Rückenmark Verletzungen (SCI) und der Motor Neuron Erkrankung, amyotrophe seitlichen Sklerose (ALS). Wir bieten ein Protokoll für die Zelle Lieferung an zervikalen Rückenmarks Vorderhorn in Nagermodellen von ALS und Rückenmarksverletzungen.

Intraspinale Cell Transplantation für Targeting Cervical Vorderhorn in Amyotrophe Lateralsklerose und traumatische Rückenmarksverletzungen

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

intraspinal-cell-transplantation-for-targeting-cervical-ventral-horn

Research

JoVE Journal

Medicine

20.5K Aufrufe.  Thomas Jefferson University Medical College. Neuronale Vorläuferzellen der Transplantation ist eine vielversprechende Strategie für den Schutz und / oder Wiederbeschaffung verlorener / dysfunktional Gebärmutterhals phrenicus Motoneuronen im Rückenmark Verletzungen (SCI) und der Motor Neuron Erkrankung, amyotrophe seitlichen Sklerose (ALS). Wir bieten ein Protokoll für die Zelle Lieferung an zervikalen Rückenmarks Vorderhorn in Nagermodellen von ALS und Rückenmarksverletzungen.

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

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.

Ein Protokoll für eine umfassende Beurteilung der bulbären Dysfunction in Amyotrophe Lateralsklerose (ALS)

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

Forschung

Medizin

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.

Akute und chronische taktile Sensorische Testung nach Verletzungen des Rückenmarks bei der Ratte

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.

Ein Modell von gequetscht Einseitige Cervical Spinal Cord Injury Mit dem 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 ...

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

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder resulting in progressive degeneration of motoneurons. Peak of onset is around 60 years for the sporadic disease and around 50 years for the familial disease. Due to its progressive course, 50% of the patients die within 30 months of symptom onset. In order to evaluate novel treatment options for this disease, genetic mouse models of ALS have been generated based on human familial mutations in the SOD gene, such as the SOD1 (G93A) mutation. Most important aspects that have to be evaluated in the model are overall survival, clinical course and motor function. Here, we demonstrate the clinical evaluation, show the conduction of two behavioural motor tests and provide quantitative scoring systems for all parameters. Because an in depth analysis of the ALS mouse model usually requires an immunohistochemical examination of the spinal cord, we demonstrate its preparation in detail applying the dorsal laminectomy method. Exemplary histological findings are demonstrated. The comprehensive application of the depicted examination methods in studies on the mouse model of ALS will enable the researcher to reliably test future therapeutic options which can provide a basis for later human clinical trials.

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

A mouse model for amyotrophic lateral sclerosis (ALS) is examined clinically and behaviorally. As a prerequisite for an accompanying immunohistological analysis the preparation of the spinal cord is depicted in detail.

Klinische Tests und Rückenmark Removal in einem Mausmodell für Amyotrophe Lateralsklerose (ALS)

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

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.

Controlled Cervical Laceration Verletzungen in Mäuse

Use of genetically modified mice enhances our  understanding of molecular mechanisms underlying several neurological disorders  such as a spinal cord ...

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

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

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

Einem Mausmodell der zervikalen Rückenmarksverletzung zu Post-läsionaler Atemneuroplastizität Studieren

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

Controlled Cortical Impact Model for Traumatic Brain Injury

Every year over a million Americans suffer a traumatic brain injury (TBI). Combined with the incidence of TBIs worldwide, the physical, emotional, social, and economical effects are staggering. Therefore, further research into the effects of TBI and effective treatments is necessary. The controlled cortical impact (CCI) model induces traumatic brain injuries ranging from mild to severe. This method uses a rigid impactor to deliver mechanical energy to an intact dura exposed following a craniectomy. Impact is made under precise parameters at a set velocity to achieve a pre-determined deformation depth. Although other TBI models, such as weight drop and fluid percussion, exist, CCI is more accurate, easier to control, and most importantly, produces traumatic brain injuries similar to those seen in humans. However, no TBI model is currently able to reproduce pathological changes identical to those seen in human patients. The CCI model allows investigation into the short-term and long-term effects of TBI, such as neuronal death, memory deficits, and cerebral edema, as well as potential therapeutic treatments for TBI.

Traumatic brain injuries (TBIs) remain a serious health problem. Using the controlled cortical impact surgery model, research on the effects of TBI and possible treatment methods may be performed.

Controlled Cortical Impact-Modell für traumatische Hirnverletzung

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.

Ein Radio-Telemetrie-System zur Herz-Kreislauf-Funktion bei Ratten mit Rückenmarksdurchtrennung und embryonalen Neural Stem Cell Grafts überwachen

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.

Synergetische Nutzung von neuralen Vorläuferzellen und Self-Montage Peptide 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. ...

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

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

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

Neural Stem Cell Transplantation in Experimental Gequetscht Modell von Rückenmarksverletzungen

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