Name: sagittal plane kinematic gait analysis in c57bl/6 mice subjected to mog35-55 induced experimental autoimmune encephalomyelitis
Uploaded: 2017-11-04T13:00:00
Description: Watch this Scientific Journal Video about Sagittal Plane Kinematic Gait Analysis in C57BL/6 Mice Subjected to MOG35-55 Induced Experimental Autoimmune Encephalomyelitis at JoVE.com

Wir würden gerne Sid Chedrawe für seine technische Unterstützung bei Dreharbeiten zu bestätigen. Diese Arbeit wurde unterstützt durch die Finanzierung von der MS Society of Canada (EGID 2983).

dieses Protokoll des Canadian Council on Animal Care Richtlinien entspricht und von der Dalhousie University Ausschuss an Labortieren genehmigt wurde.
 1. konstruieren reflektierenden Markierungen: <ol><li>mit einem handgeführten Locher, die gewünschte Anzahl von kleinen Kreisen von reflektierenden Papier Stanzen. Jedes Tier benötigt 5 Marker für eine einzelne Aufnahme; zwei große und drei kleine Markierungen.</li>
	<li>Mit einer feinen Schere, einen geraden Schnitt erstreckt sich von ...

<ol>
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(91), e51785 (2014).</li><li>Hetze, S., Romer, C., Teufelhart, C., Meisel, A., Engel, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gait+analysis+as+a+method+for+assessing+neurological+outcome+in+a+mouse+model+of+stroke.">Gait analysis as a method for assessing neurological outcome in a mouse model of stroke.</a> J Neurosci Methods. 206 (1), 7-14 (2012).</li><li>Preisig, D. F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-speed+video+gait+analysis+reveals+early+and+characteristic+locomotor+phenotypes+in+mouse+models+of+neurodegenerative+movement+disorders.">High-speed video gait analysis reveals early and characteristic locomotor phenotypes in mouse models of neurodegenerative movement disorders.</a> Behav Brain Res. 311, 340-353 (2016).</li><li>Leblond, H., L'Esperance, M., Orsal, D., Rossignol, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Treadmill+locomotion+in+the+intact+and+spinal+mouse.">Treadmill locomotion in the intact and spinal mouse.</a> J Neurosci. 23 (36), 11411-11419 (2003).</li><li>Ueno, M., Yamashita, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kinematic+analyses+reveal+impaired+locomotion+following+injury+of+the+motor+cortex+in+mice.">Kinematic analyses reveal impaired locomotion following injury of the motor cortex in mice.</a> Exp Neurol. 230 (2), 280-290 (2011).</li><li>Zorner, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Profiling+locomotor+recovery:+comprehensive+quantification+of+impairments+after+CNS+damage+in+rodents.">Profiling locomotor recovery: comprehensive quantification of impairments after CNS damage in rodents.</a> Nat Methods. 7 (9), 701-708 (2010).</li><li>Pearson, K. G., Acharya, H., Fouad, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+electrode+configuration+for+recording+electromyographic+activity+in+behaving+mice.">A new electrode configuration for recording electromyographic activity in behaving mice.</a> J Neurosci Methods. 148 (1), 36-42 (2005).</li><li>Balkaya, M., Krober, J. M., Rex, A., Endres, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessing+post-stroke+behavior+in+mouse+models+of+focal+ischemia.">Assessing post-stroke behavior in mouse models of focal ischemia.</a> J Cereb Blood Flow Metab. 33 (3), 330-338 (2013).</li><li>Akay, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-term+measurement+of+muscle+denervation+and+locomotor+behavior+in+individual+wild-type+and+ALS+model+mice.">Long-term measurement of muscle denervation and locomotor behavior in individual wild-type and ALS model mice.</a> J Neurophysiol. 111 (3), 694-703 (2014).</li><li>Taylor, T. N., Greene, J. G., Miller, G. W. <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+of+mouse+models+of+Parkinson's+disease.">Behavioral phenotyping of mouse models of Parkinson's disease.</a> Behav Brain Res. 211 (1), 1-10 (2010).</li><li>Chen, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+Histopathological+and+Behavioral+Outcomes+Eight+Weeks+after+Rat+Spinal+Cord+Injury+by+Contusion,+Dislocation,+and+Distraction+Mechanisms.">Differential Histopathological and Behavioral Outcomes Eight Weeks after Rat Spinal Cord Injury by Contusion, Dislocation, and Distraction Mechanisms.</a> J Neurotrauma. 33 (18), 1667-1684 (2016).</li><li>de Bruin, N. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multiple+rodent+models+and+behavioral+measures+reveal+unexpected+responses+to+FTY720+and+DMF+in+experimental+autoimmune+encephalomyelitis.">Multiple rodent models and behavioral measures reveal unexpected responses to FTY720 and DMF in experimental autoimmune encephalomyelitis.</a> Behav Brain Res. 300, 160-174 (2016).</li><li>Steinman, L., Zamvil, S. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=How+to+successfully+apply+animal+studies+in+experimental+allergic+encephalomyelitis+to+research+on+multiple+sclerosis.">How to successfully apply animal studies in experimental allergic encephalomyelitis to research on multiple sclerosis.</a> Ann Neurol. 60 (1), 12-21 (2006).</li><li>Emerson, M. R., Gallagher, R. J., Marquis, J. G., LeVine, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enhancing+the+ability+of+experimental+autoimmune+encephalomyelitis+to+serve+as+a+more+rigorous+model+of+multiple+sclerosis+through+refinement+of+the+experimental+design.">Enhancing the ability of experimental autoimmune encephalomyelitis to serve as a more rigorous model of multiple sclerosis through refinement of the experimental design.</a> Comp Med. 59 (2), 112-128 (2009).</li><li>Bittner, S., Afzali, A. M., Wiendl, H., Meuth, S. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Myelin+oligodendrocyte+glycoprotein+(MOG35-55)+induced+experimental+autoimmune+encephalomyelitis+(EAE)+in+C57BL/6+mice.">Myelin oligodendrocyte glycoprotein (MOG35-55) induced experimental autoimmune encephalomyelitis (EAE) in C57BL/6 mice.</a> J Vis Exp. (86), (2014).</li><li>Beeton, C., Garcia, A., Chandy, K. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+and+clinical+scoring+of+chronic-relapsing+experimental+autoimmune+encephalomyelitis.">Induction and clinical scoring of chronic-relapsing experimental autoimmune encephalomyelitis.</a> J Vis Exp. (5), e224 (2007).</li><li>Barthelmes, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+Experimental+Autoimmune+Encephalomyelitis+in+Mice+and+Evaluation+of+the+Disease-dependent+Distribution+of+Immune+Cells+in+Various+Tissues.">Induction of Experimental Autoimmune Encephalomyelitis in Mice and Evaluation of the Disease-dependent Distribution of Immune Cells in Various Tissues.</a> J Vis Exp. (111), (2016).</li><li>Shaw, M. K., Zhao, X. Q., Tse, H. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Overcoming+unresponsiveness+in+experimental+autoimmune+encephalomyelitis+(EAE)+resistant+mouse+strains+by+adoptive+transfer+and+antigenic+challenge.">Overcoming unresponsiveness in experimental autoimmune encephalomyelitis (EAE) resistant mouse strains by adoptive transfer and antigenic challenge.</a> J Vis Exp. (62), e3778 (2012).</li><li>Stromnes, I. M., Goverman, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Passive+induction+of+experimental+allergic+encephalomyelitis.">Passive induction of experimental allergic encephalomyelitis.</a> Nat Protoc. 1 (4), 1952-1960 (2006).</li><li>Stromnes, I. M., Goverman, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Active+induction+of+experimental+allergic+encephalomyelitis.">Active induction of experimental allergic encephalomyelitis.</a> Nat Protoc. 1 (4), 1810-1819 (2006).</li><li>Fiander, M. D., Stifani, N., Nichols, M., Akay, T., Robertson, G. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kinematic+gait+parameters+are+highly+sensitive+measures+of+motor+deficits+and+spinal+cord+injury+in+mice+subjected+to+experimental+autoimmune+encephalomyelitis.">Kinematic gait parameters are highly sensitive measures of motor deficits and spinal cord injury in mice subjected to experimental autoimmune encephalomyelitis.</a> Behav Brain Res. 317, 95-108 (2017).</li><li>Jones, M. V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+and+pathological+outcomes+in+MOG+35-55+experimental+autoimmune+encephalomyelitis.">Behavioral and pathological outcomes in MOG 35-55 experimental autoimmune encephalomyelitis.</a> J Neuroimmunol. 199 (1-2), 83-93 (2008).</li><li>van den Berg, R., Laman, J. D., van Meurs, M., Hintzen, R. Q., Hoogenraad, C. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rotarod+motor+performance+and+advanced+spinal+cord+lesion+image+analysis+refine+assessment+of+neurodegeneration+in+experimental+autoimmune+encephalomyelitis.">Rotarod motor performance and advanced spinal cord lesion image analysis refine assessment of neurodegeneration in experimental autoimmune encephalomyelitis.</a> J Neurosci Methods. 262, 66-76 (2016).</li><li>Sasaki, M., Lankford, K. L., Brown, R. J., Ruddle, N. H., Kocsis, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Focal+experimental+autoimmune+encephalomyelitis+in+the+Lewis+rat+induced+by+immunization+with+myelin+oligodendrocyte+glycoprotein+and+intraspinal+injection+of+vascular+endothelial+growth+factor.">Focal experimental autoimmune encephalomyelitis in the Lewis rat induced by immunization with myelin oligodendrocyte glycoprotein and intraspinal injection of vascular endothelial growth factor.</a> Glia. 58 (13), 1523-1531 (2010).</li><li>Merkler, D., Ernsting, T., Kerschensteiner, M., Bruck, W., Stadelmann, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+focal+EAE+model+of+cortical+demyelination:+multiple+sclerosis-like+lesions+with+rapid+resolution+of+inflammation+and+extensive+remyelination.">A new focal EAE model of cortical demyelination: multiple sclerosis-like lesions with rapid resolution of inflammation and extensive remyelination.</a> Brain. 129 (Pt 8), 1972-1983 (2006).</li><li>Franco-Pons, N., Torrente, M., Colomina, M. 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Bei Mäusen mit EAE die zwei am häufigsten verwendeten Methoden zur Messung der motorischen Defizite sind klinische Bewertung und Latenz von einer Rotarod27,28fallen. Diese Techniken haben einige Einschränkungen. Obwohl praktisch und weit verbreitet, beschränkt klinische Bewertung durch nachgeben nur ordinale Daten, was bedeutet, dass das Ausmaß der Unterschiede zwischen klinischen Resultate sind nicht bekannt. Klinische Bewertung leidet auch nicht in der Lag...

Gang ist eine Reihe von sich wiederholenden Bewegungen der Gliedmaßen zur Fortbewegung zu erreichen. Gang besteht aus Schritt Zyklen, die in zwei Phasen unterteilt werden: die Standphase, was ist, wenn der Fuß auf dem Boden nach hinten bewegt, um treiben den Körper nach vorne; und die Schwungphase, wo der Fuß aus den Boden und bewegen nach vorne ist. Störungen des Gehens sind die typischen Merkmale der vielen neurodegenerativen Erkrankungen, wie z. B. Verletzungen des Rückenmarks (SCI), Multiple Sklerose (MS), Amyotrophe Lateralsklerose (ALS), Parkinson-Krankheit (PD) und Schlaganfall; präklinischen Nager-Modelle dieser Störungen rekapitulieren oft ihre jeweiligen Gangart Beeinträchtigungen1. Die grundlegende Kontrollmechanismen der Fortbewegung bei Mäusen wurden intensiv studiert2,3. Darüber hinaus gibt es Maus-Modellen von vielen menschlichen neurologische Störungen4. Ganganalyse bei Mäusen ist daher ein attraktiver Ansatz, mehrere Aspekte der motorischen Defizite zu messen, die anatomische Korrelate bekannt haben. Die Studie des Gehens in Mausmodellen kann Einblicke in die neuropathologische Grundlagen der motorischen Defizite bei neurodegenerativen Erkrankungen und ermöglicht die Identifizierung von möglichen Behandlungen.
Einige Techniken, die zur Messung der Gangart bei Nagetieren gehören Sichtprüfung (z.B., Basso Maus Maßstab5 und Freiland Test6) und Analyse der Gang von der ventralen Flugzeug-7. Methoden zur Sagittalebene Kinematik der Megalosauridae Bewegungen Messen haben vor kurzem Popularität gewonnen, weil sie mehr über die Ausführung der Bewegung Informationen und sind daher empfindlicher auf subtile Veränderungen in Gang8, 9 , 10 , 11. kinematische Techniken entwickelt, um die Megalosauridae Bewegung in der Sagittalebene studieren beim gehen auf einem Laufband-9,-12 wurden im Rahmen des SCI, ALS, kortikale traumatische Verletzungen, Schlaganfall, ausgiebig untersucht und Chorea Huntington8,9,10,11,13,14,15,16. Im Gegensatz dazu haben diese Techniken eingeschränkte Nutzung in der Studie der motorischen Defizite für Maus-Modellen von Multipler Sklerose17gesehen.
Experimentelle autoimmune Enzephalomyelitis (EAE) ist die am häufigsten verwendeten Mausmodell der MS18. Die zwei wichtigsten Methoden des Verursachens EAE erfolgt über aktive oder passive Impfung. In aktive EAE sind Mäuse mit Myelin Antigene, verursachen autoreaktiven T-Zell-vermittelten Neuroinflammation und Demyelinisierung im Rückenmark und Kleinhirn immunisiert. Passive EAE, wird auf der anderen Seite durch Übertragung von autoreaktiven T-Zellen von einer Maus mit aktive EAE auf einen naiven Maus19induziert. Wie an anderer Stelle beschrieben, den Krankheitsverlauf und Neuropathologie sind beeinflusst durch das zentrale Nervensystem (ZNS) Antigen und Maus belasten20,21,22,23,24 ,25. Bei der EAE Experimente sind Kontrollmäusen komplette Freund des Adjuvans (CFA) ohne das Myelin-Antigen gespritzt. EAE zeichnet sich durch aufsteigende Lähmungen, die beginnt mit Schweif Schwäche und kann potentiell die Vorderbeine mit Ataxie und Lähmung20führt. Wir haben vor kurzem Gang Änderungen bei C57Bl/6 Mäusen unterworfen Myelin Oligodendrozyt Glykoprotein 35-55 (MOG35-55) gekennzeichnet-induzierte EAE. Diese Studien haben gezeigt, dass Ganganalyse als klassische Verhaltensanalyse überlegen sein, denn Abweichungen vom normalen Knöchel Bewegung mit dem Grad der weißen Substanz-Verlust in der Lendenwirbelsäule Rückenmark von EAE Mäusen26stark korreliert sind. Im Gegensatz dazu war die Stärke der Korrelation zwischen weißen Substanz-Verlust und zwei andere traditionelle Verhaltensmaßnahmen (klinische scoring und Rotarod) viel schwächeren26.
Wir beschreiben hier die Verwendung von kinematischen Ganganalyse, Bewegung Defizite in der Sagittalebene von EAE Mäusen gehen auf einem Laufband zu erkennen. Fünf reflektierende Markierungen wurden auf eine Megalosauridae Bewegung der Hüfte, Knie und Sprunggelenke in High-Speed-Videoaufnahmen identifizieren gesetzt. Bewegungsanalyse Software wurde verwendet, um kinematische Daten über gemeinsame Ausflüge zu extrahieren. Das Dienstprogramm dieser Techniken zu quantifizieren, Bewegung Defizite für das MOG-35-55 -Modell der EAE werden diskutiert. Diese Techniken sind auch für das Studium der Gangart Defizite in anderen Maus-Modellen von neurodegenerativen Erkrankungen.

<table border="0" cellpadding="0" cellspacing="0" width="893">
	<tbody>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Camera</td>
			<td style="width:167px;">Nikon</td>
			<td style="width:344px;">Nikon D750</td>
			<td style="width:167px;">Used to film the video</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Reflective tape</td>
			<td>B&#38;L Engineering</td>
			<td>MKR-Tape-2</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Fine scissors</td>
			<td>Fine Science Tools</td>
			<td>15023-10</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Forceps</td>
			<td>Fine Science Tools</td>
			<td>11252-20</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Glue gun</td>
			<td>Craftsmart</td>
			<td>E231647</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">scalpel handle #4</td>
			<td>Roboz</td>
			<td>R5-9884</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Scalpel Blade No.10</td>
			<td>Feather</td>
			<td>2020-12</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">C57BL/6 mice</td>
			<td>Charles River Laboratories</td>
			<td style="width:344px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Anesthetic machine</td>
			<td>EZ Anesthesia</td>
			<td>EZ-AF9000 Auto Flow System</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Recirculating water heating blanket</td>
			<td>Androit</td>
			<td>HTP-1500</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">topical eye lubricant</td>
			<td>Refresh</td>
			<td>DIN00210889</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Shaver</td>
			<td>Oster</td>
			<td>78997-010</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">High speed camera</td>
			<td>Fastec</td>
			<td>Fastec IL3-100</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">High power light</td>
			<td>Smith Victor Corporation</td>
			<td>Model 700 SG (600 Watt quartz light, 120 Volts)</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Light Stand</td>
			<td>Promaster</td>
			<td>LS1</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Treadmill</td>
			<td colspan="2">Custom built at the Zoological Institute, University of Cologne</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Microsoft Excel 2016</td>
			<td>Microsoft</td>
			<td>Version 2016</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">KinemaJ</td>
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sagittal plane kinematic gait analysis in c57bl/6 mice subjected to mog35-55 induced experimental autoimmune encephalomyelitis

Kinematische Ganganalyse in der Sagittalebene wurde häufig verwendet, um motorische Defizite bei Multipler Sklerose (MS) zu charakterisieren. Wir beschreiben die Anwendung dieser Techniken, Gang Defizite in einem Mausmodell der MS, bekannt als experimentelle autoimmune Enzephalomyelitis (EAE) zu identifizieren. Lähmung und Motor Defizite bei Mäusen EAE ausgesetzt sind in der Regel anhand einer klinischen Bewertungen Skala. Dieser Größenordnung führt jedoch nur ordinale Daten wenig Informationen über die genaue Art der motorischen Defizite. EAE Krankheitschwierigkeit wurde auch durch Rotarod Leistung bewertet stellt ein Maß für die allgemeine motorische Koordination. Im Gegensatz dazu erzeugt kinematische Ganganalyse der hinteren Extremität in der Sagittalebene höchst präzise Informationen darüber, wie die Bewegung beeinträchtigt ist. Um dieses Verfahren ausführen zu können, sind reflektierende Markierungen auf Hind Gliedmaßen, Beweglichkeit der Gelenke zu erkennen, während eine Maus auf einem Laufband Fuß gelegt. Bewegungsanalyse Software wird verwendet, um die Bewegung der Marker während des Gehens zu messen. Kinematische Gangartparameter werden dann aus den daraus resultierenden Daten abgeleitet. Wir zeigen, wie diese Gangartparameter verwendet werden können, beeinträchtigte Bewegungen der Hüfte, Knie und Knöchel Gelenke in EAE zu quantifizieren. Diese Techniken können verwendet werden, zum besseren Verständnis der Krankheitsmechanismen und identifizieren mögliche Behandlungen für MS und anderen neurodegenerativen Erkrankungen, die Mobilität zu beeinträchtigen.

Abbildung 1 zeigt eine schematische Darstellung des Verfahrens für kinematische Ganganalyse verwendet. Erste, reflektierende Markierungen gemacht und auf eine Maus an 5 anatomischen Punkten platziert. Gang wird dann aufgezeichnet, während die Maus auf einem Laufband Fuß ist. Bewegungsanalyse Software wird verwendet, um kinematische Daten für spätere Analysen zu extrahieren.
Abbildung 2A -C repräsentieren die Sch...

Watch this Scientific Journal Video about Sagittal Plane Kinematic Gait Analysis in C57BL/6 Mice Subjected to MOG35-55 Induced Experimental Autoimmune Encephalomyelitis at JoVE.com

Sagittal Plane Kinematic Gait Analysis in C57BL/6 Mice Subjected to MOG35-55 Induced Experimental Autoimmune Encephalomyelitis

Sagittalebene kinematische Ganganalyse bei C57BL/6 Mäusen unterworfen MOG35-55 induzierte experimentelle Autoimmune Enzephalomyelitis

Kinematische Ganganalyse in der Sagittalebene liefert präzise Informationen über wie Bewegung ausgeführt wird. Wir beschreiben die Anwendung dieser Techniken, Gang Defizite für Mäuse ausgesetzt Autoimmun-vermittelter Demyelinisierung zu identifizieren. Diese Methoden können auch verwendet werden, Gang Defizite für andere Maus-Modellen mit eingeschränkter Fortbewegung zu charakterisieren.

Kinematic gait analysis in the sagittal plane has frequently been used to characterize motor deficits in multiple sclerosis (MS). We describe the ...

The overall goal of this procedure is to measure functional motor impairments in the experimental autoimmune encephalomyelitis or EAE, mouse model multiple sclerosis using kinematic gait analysis. The application of kinematic gait analysis to mouse-walking behavior has been previously established and described by others. Neurological deficits in an EAE result from neural inflammation and sporadic white matter loss throughout the spinal cord and the cerebellum.Traditionally, motor stability in EAE mice has been assessed using clinical scoring systems, in which mice are assigned a clinical score based on the observer's impression of the severity of motor deficits. Data from clinical scores are ordinal and do not correlate well with the spinal cord histopathology. Kinematic gait analysis has recently been shown to be a better behavioral correlate of white matter loss than clinical scores, in addition to providing an objective description of walking deficits in the EAE mice.This technique involves placing reflective markers on the high limbs of mice and allowing them to walk on a treadmill while being recorded with a high-speed camera. Kinematic parameters are then extracted from the video using motion analysis software. The first step is to make the markers that will be placed on the animal's hind leg.The reflection of light off of these markers allows the coordinates of anatomical points on the leg to be extracted from the videos. To begin, punch the desired number of circles from the sheet of reflective paper. Each mouse requires five markers per recording, two large and three small.Using fine scissors, make a cut extending from the perimeter to the center of the circle. Peel off the paper backing to reveal the adhesive surface using fine forceps. Grip the marker with forceps and using your finger, roll the marker in on itself to form a cone.To make a small marker, make a longer cut and curl the cone more tightly. To make a large marker, make a shorter cut and curl the cone more loosely. Using a hot glue gun, fill the inside of the cone with glue and adhere it to a piece of cardboard.It is important to fully fill the marker with glue to prevent the marker from collapsing when handling during recording. Once the glue is dry, cut the marker off using a scalpel. Ensure that you cut away from your body.The next step is to prepare the animal for recording. To do this, the reflective markers must be adhered to the hind limb of the mouse at the proper anatomical locations. This is conducted under light anesthesia.Place the mouse in an induction chamber and anesthetize with 2.5%isoflurane. Once unconscious, transfer the mouse from the induction chamber to a nose cone or a recirculating water heating pad. Apply a topical lubricant to both eyes.Shave the desired hind limb extending from the ankle to the spine and the bottom of the ribs. In this demonstration, we are recoding movements of the right hind limb, but either or both limbs may be used. Locate the iliac crest by bringing both knees together with your thumb and forefinger and palpating just below the ribs.Mark this location using a permanent marker. Locate the hip by extending the leg and moving it back and forth. Place a marker over the hip joint, which is the point of articulation between the head of the femur and the pelvis.Using a flexible ruler, measure and record the length of the tibia or shank of the mouse. Measure and record the length of the femur or thigh as well. To isolate the fourth toe for marker placement, tape down the rest of the foot.Grasp a small marker with forceps and dip the flat end in glue. Place the small marker on the tip of the fourth toe. Place another small marker on the metatarsophalangeal joint.Place the last small marker on the ankle. Place a large marker on the hip joint directly over the mark on the skin. Place the second large marker on the mark over the iliac crest.Remove the tape from the foot. Place the mouse in a recovery cage and transport immediately to the gait recording room. The next step is recording the mouse walking on a treadmill.This picture is of our treadmill recording setup, showing the light, high-speed camera, and treadmill. Prior to recording the mouse's gait, take a picture of a calibration block with known dimensions on the treadmill. This will allow the pixels in the video to be converted to real measurements.It is critical that the camera angle and position at which the calibration picture is taken remains the same during the recording of walking behavior. Place the mouse in the treadmill. Increase the treadmill's speed gradually to orient the mouse in the correct direction.Slowly accelerate to 20 centimeters per second, which is the ideal running speed for obtaining recordings of steady gait. For accurate analysis, it's best to record eight to 12 step cycles of steady walking behavior. This is an example of consistent walking.The following is the same video shown at half speed. Because mice walk so quickly, it can be useful to view the videos at slower speeds to count step cycles and to better appreciate the gait pattern. This is one example of an EAE mouse walking on the treadmill.This mouse is unable to support its body weight as its pelvis is very low to the ground. Additionally, it is having difficulty lifting its foot off the ground during the swing phase. This is an example of another EAE mouse that has less severe motor deficits.The EAE mouse is walking on its toes with reduced movement at the ankle joint. This gives the mouse an uneven gait. The following are examples of behaviors that will compound analysis.Recordings, including these behaviors, should not be used. Lagging occurs when the mouse stops walking and travels to the back of the treadmill, but then resumes walking. This may occur in any mouse, but will occur more frequently at lower speeds.Rearing occurs when the mouse shifts its weight on its back hind limbs and raises its head and upper body. This behavior is common in anxious mice. The following are examples of poor lighting that will lead to issues with analysis of the markers'position in time and space.If the lighting is too dim, markers may not reflect enough light to be recognized by the computer software. In this video, the toe markers are not easily visible due to insufficient lighting. If the lighting is too bright, objects other than the markers may reflect too much light and be recognized as a marker.In this video, the toe markers appear to be merging with the reflective parts of the treadmill. Data from the reflective markers on mice walking can be used to create stick models of the leg from which kinematic parameters can be extracted. This is a recording of a mouse walking with a stick model of the leg superimposed.Note that in this video, there is a sixth marker on the knee, which is not necessary because the location of the knee can be triangulated from the positions of the hip and ankle joints, and the measured lengths of the femur and tibia. Because markers over the knee joint are often inaccurate due to skin slippage, triangulation is the preferred method. A mouse's step cycle can be divided into two main phases.The stance phase and the swing phase. This stick diagram can be used to qualitatively illustrate the movement of the hind limb in time and space. An example of this could be to assess the movement of the hind limb at different times points throughout the study.This example shows that the hind leg is compressed during the stance phase, indicating that the mouse has difficulty supporting its body weight. This is reflected by increased flexion of the knee and ankle joints. At the later time point, this has partially recovered.This video illustrates the relationship between the walking behavior and joint angles over time. The hip, knee, and ankle angle waveforms can be extracted from each recording. The joint angles can then be averaged over eight to 12 consecutive step cycles, resulting in an average step cycle that can be used for further analysis.This graph represents a knee joint waveform averaging from 10 consecutive step cycles. The data has been normalized, so that the length of the stance and swing phases are 100 frames respectively. The clear background represents the swing phase and the green background represents the stance phase.For this mouse, the walking behavior from week to week was very consistent and the step-cycle waveforms from each week overlapped considerably. However, there can also be substantial variability in gait in healthy mice as seen in this graph. The degree of variability shown here is acceptable and within the range of what might be expected from a mouse.This graph represents the knee step cycle from the EAE mouse recorded on three consecutive weeks. There is a slight change in the shape of the step cycle at week two and a substantial deviation by week three, which the mouse's knee is much more flexed and does not extend during walking. There are numerous parameters that can be measured using this technique.We'll briefly describe three. The average angle is obtained by averaging all angles throughout the normalized step cycle. In this instance, the average angle decreases throughout the study, suggesting that the mice are not extending their knees as much as normal.The range of motion is obtained by subtracting the smallest angle from the largest angle in the normalized step cycle. This parameter can give you insight into joint flexibility, rigidity, or weakness. In this example, the knee range of motion decreases throughout the study indicating that the mice are not able to move their knee normally, possibly due to muscle weakness.Root-Mean-Square difference is a method used to measure deviation of step-cycle waveforms from the baseline recording. This parameter tells you how much deviation there is from the initial recording. Kinematic gait analysis is a valuable technique that can be used to sensitively detect and describe changes in gait.The application of kinematic gait analysis to EAE studies may be a valuable tool in understanding the functional consequences of spinal cord pathology in this model. It may facilitate discovery of novel treatments for multiple sclerosis. Additionally, kinematic gait analysis is not limited to the context of EAE.This technique has previously been used in mouse models with spinal cord injury, amyotrophic lateral sclerosis, Huntington's disease and stroke, and can be applied to other mouse models with neurological disorders, including Parkinson's disease.

Research

JoVE Journal

Neuroscience

Experimental Methods for Testing the Effects of Neurotrophic Peptide, ADNF-9, Against Alcohol-induced Apoptosis during Pregnancy in C57BL/6 Mice

Experimentelle Methoden zur Prüfung der Auswirkungen von Neurotrophic Peptide, ADNF-9, gegen den Alkohol-induzierte Apoptose während der Schwangerschaft in C57BL / 6 Mäuse

Experimental designs for investigating the effects of prenatal alcohol exposure during early embryonic stages in fetal brain growth are challenging. This ...

Forschung

Neurowissenschaften

Automated Gait Analysis in Mice with Chronic Constriction Injury

Automatisierte Ganganalyse bei Mäusen mit chronischen Verengung Verletzungen

The von Frey test is a classical method that has been widely used to examine the sensory function of neuropathic pain animals. However, it has some ...

A Simple Approach to Induce Experimental Autoimmune Neuritis in C57BL/6 Mice for Functional and Neuropathological Assessments

Ein einfacher Ansatz, experimentelle Autoimmune Neuritis bei C57BL/6 Mäusen für funktionale und neuropathologische Bewertungen zu induzieren

Experimental autoimmune neuritis (EAN) is a well-appreciated experimental model of autoimmune peripheral demyelinating diseases. EAN disease is induced by ...

Cochlear Implant Surgery and Electrically-evoked Auditory Brainstem Response Recordings in C57BL/6 Mice

Cochlea-Implantat-Chirurgie und elektrisch evozierten Auditory Brainstem Response Aufnahmen bei C57BL/6 Mäusen

Cochlear implants (CIs) are neuroprosthetic devices that can provide a sense of hearing to deaf people. However, a CI cannot restore all aspects of ...

Comprehensive Understanding of Inactivity-Induced Gait Alteration in Rodents

Umfassendes Verständnis der inaktivitätsinduzierten Gangveränderung bei Nagetieren

It is well known that disuse affects neural systems and that joint motions become altered; however, which outcomes properly exhibit these characteristics ...

Imaging CD19+ B Cells in an Experimental Autoimmune Encephalomyelitis Mouse Model using Positron Emission Tomography

Imaging CD19+ B Cells in an Experimental Autoimmune Encephalomyelitis Mouse Model using Positron Emission Tomography

Bildgebung von CD19+ B-Zellen in einem experimentellen autoimmunen Enzephalomyelitis-Mausmodell mit Positronen-Emissions-Tomographie

Multiple sclerosis (MS) is the most common demyelinating central nervous system (CNS) disease affecting young adults, often resulting in neurological ...

Induktion und Evaluation der experimentellen autoimmunen Enzephalomyelitis bei der Maus

Modeling Multiple Sclerosis in the Two Sexes: MOG35-55-Induced Experimental Autoimmune Encephalomyelitis

Autor im Rampenlicht: Entwicklung eines vielseitigen experimentellen Modells für Autoimmun-Enzephalomyelitis, das sowohl für männliche als auch für weibliche Mäuse relevant ist 

Modellierung der Multiplen Sklerose bei beiden Geschlechtern: MOG35-55-induzierte experimentelle Autoimmunenzephalomyelitis

Multiple Sclerosis (MS) is a chronic autoimmune inflammatory disease affecting the central nervous system (CNS). It is characterized by different ...

Soziales Stressparadigma bei Jugendlichen bei C57BL/6-Mäusen für die Verhaltensforschung

Social Defeat Stress Model for Adolescent C57BL&#47;6 Male and Female Mice

Autor im Rampenlicht: Verständnis der Auswirkungen sozialer Widrigkeiten bei Jugendlichen auf die Neuroentwicklung bei Mäusen

Soziales Defeat Stress-Modell für jugendliche C57BL/6 männliche und weibliche Mäuse

Social adversity in adolescence is prevalent and can negatively impact mental health trajectories. Modeling social stress in adolescent male and female ...