Name: automated gait analysis in mice with chronic constriction injury
Uploaded: 2017-10-17T14:00:00
Description: Watch this Scientific Journal Video about Automated Gait Analysis in Mice with Chronic Constriction Injury at JoVE.com

Denne forskningen ble støttet av Chungnam National University, Korea Institute av orientalsk medisin (KIOM) og stipend til Korea helse teknologi R &#38; D prosjektet gjennom den Korea helse industri Development Institute (KHIDI), finansiert av Helsedepartementet &#38; Velferd, Sør-Korea (gi nummer: HI15C0007).

alle eksperimentene ble utført i samsvar med etiske retningslinjer International Association for studier av smerte og ble godkjent av institusjonelle Animal Care og bruk komiteen på Chungnam National University ( Daejeon, Sør-Korea).
 1. induksjon av CCI på Sciatic Nerve <ol><li>huset mannlige ICR mus veier 20-25 g under en 12t lys/mørke syklus i kontrollerte romtemperatur (opprettholdt på 20-25 ° C) og luftfuktighet (40-60%), med gratis tilgang til mat og vann. Tillate en ac...

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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+peripheral+mononeuropathy+in+rat+that+produces+disorders+of+pain+sensation+like+those+seen+in+man.">A peripheral mononeuropathy in rat that produces disorders of pain sensation like those seen in man.</a> Pain. 33 (1), 87-107 (1988).</li><li>Dixon, W. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient+analysis+of+experimental+observations.">Efficient analysis of experimental observations.</a> Annu Rev Pharmacol Toxicol. 20, 441-462 (1980).</li><li>Bonin, R. P., Bories, C., De Koninck, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simplified+up-down+method+(SUDO)+for+measuring+mechanical+nociception+in+rodents+using+von+Frey+filaments.">A simplified up-down method (SUDO) for measuring mechanical nociception in rodents using von Frey filaments.</a> Mol Pain. 10, 26 (2014).</li><li>Chaplan, S. R., Bach, F. W., Pogrel, J. W., Chung, J. M., Yaksh, T. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+assessment+of+tactile+allodynia+in+the+rat+paw.">Quantitative assessment of tactile allodynia in the rat paw.</a> J Neurosci Methods. 53 (1), 55-63 (1994).</li><li>Chen, H., Du, J., Zhang, Y., Barnes, K., Jia, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Establishing+a+Reliable+Gait+Evaluation+Method+for+Rodent+Studies.">Establishing a Reliable Gait Evaluation Method for Rodent Studies.</a> J Neurosci Methods. , (2017).</li><li>Kang, D. W., 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=Antinociceptive+Profile+of+Levo-tetrahydropalmatine+in+Acute+and+Chronic+Pain+Mice+Models:+Role+of+spinal+sigma-1+receptor.">Antinociceptive Profile of Levo-tetrahydropalmatine in Acute and Chronic Pain Mice Models: Role of spinal sigma-1 receptor.</a> Sci Rep. 6, 37850 (2016).</li><li>Huehnchen, P., Boehmerle, W., 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=Assessment+of+paclitaxel+induced+sensory+polyneuropathy+with+&amp;#34;Catwalk&amp;#34;+automated+gait+analysis+in+mice.">Assessment of paclitaxel induced sensory polyneuropathy with &amp;#34;Catwalk&amp;#34; automated gait analysis in mice.</a> PLoS One. 8 (10), e76772 (2013).</li><li>Vrinten, D. H., Hamers, F. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term='CatWalk'+automated+quantitative+gait+analysis+as+a+novel+method+to+assess+mechanical+allodynia+in+the+rat;+a+comparison+with+von+Frey+testing.">'CatWalk' automated quantitative gait analysis as a novel method to assess mechanical allodynia in the rat; a comparison with von Frey testing.</a> Pain. 102 (1-2), 203-209 (2003).</li><li>Martinov, T., Mack, M., Sykes, A., Chatterjea, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measuring+changes+in+tactile+sensitivity+in+the+hind+paw+of+mice+using+an+electronic+von+Frey+apparatus.">Measuring changes in tactile sensitivity in the hind paw of mice using an electronic von Frey apparatus.</a> J Vis Exp. (82), e51212 (2013).</li><li>Ferland, C. E., Laverty, S., Beaudry, F., Vachon, P. <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+and+pain+response+of+two+rodent+models+of+osteoarthritis.">Gait analysis and pain response of two rodent models of osteoarthritis.</a> Pharmacol Biochem Behav. 97 (3), 603-610 (2011).</li><li>Mogil, J. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hypolocomotion,+asymmetrically+directed+behaviors+(licking,+lifting,+flinching,+and+shaking)+and+dynamic+weight+bearing+(gait)+changes+are+not+measures+of+neuropathic+pain+in+mice.">Hypolocomotion, asymmetrically directed behaviors (licking, lifting, flinching, and shaking) and dynamic weight bearing (gait) changes are not measures of neuropathic pain in mice.</a> Mol Pain. 6, 34 (2010).</li><li>Ferreira-Gomes, J., Adaes, S., Castro-Lopes, 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=Assessment+of+movement-evoked+pain+in+osteoarthritis+by+the+knee-bend+and+CatWalk+tests:+a+clinically+relevant+study.">Assessment of movement-evoked pain in osteoarthritis by the knee-bend and CatWalk tests: a clinically relevant study.</a> J Pain. 9 (10), 945-954 (2008).</li></ol>

Måling av mekaniske allodynia ved hjelp av von Frey filamenter er foreløpig mest brukte metoden i smerte dyremodeller for å demonstrere taktile overfølsomhet. Som dyr modeller for nevropatisk smerte har fortsatt å bli utviklet, er metodikk vurdering for sensorisk funksjonen også forbedret8,9,10,15. I disse rapportene, har det blitt foreslått disse endringene kan gi en mer nøyaktig, rask...

Patologiske forandringer av nervesystemet indusert av traumer, metabolske dysfunksjon, betennelse, infeksjon, iskemi, eller autoimmune sykdommer noen ganger føre til nevropatisk smerte, som er definert som en smerte som oppstår som en direkte konsekvens av en lesjon eller sykdom påvirker somatosensory system1. Dessverre konvensjonelle analgetika vanligvis produserer ikke tilstrekkelig smerte lindring2nevropatisk smerte er vanligvis uutholdelig. En viktig funksjon i nevropatisk smerte inneholder spontan og stimulering-utløste (dvs., allodynia og hyperalgesia) smerter. Allodynia er et nociceptive svar som oppstår vanligvis ikke-smertefulle stimuli, som lett berøring eller varm stimulering. Hyperalgesia angir et forbedret smerte svar til skadelige mekanisk og/eller termisk stimuli3. Selv om disse to symptomer både kritisk svekke pasientens livskvalitet, er mekanisk allodynia fremkalt av mild taktil stimulering mest skjerpende symptomet fordi myk kontakt er vanskelig å unngå i hverdagen.
Undersøke den underliggende mekanismen og nye smertestillende for behandling av nevropatisk smerte, nøyaktig måling av smerte svaret er viktig. Mange nevropatisk smerte dyremodeller har utviklet nociceptive svar på hind pote området på grunn av dens høy tilgjengelighet4,5,6,7. Dermed har mest smerte svar vurderinger blitt utført på plantar eller dorsal overflaten av bakben labben ved å bruke mekaniske stimuli spesielle instrumenter, som von Frey filamenter. En av de mest brukte metoden er opp ned beskrevet av Dixon8 og senere endret versjoner9,10. Men veldig dyktig, erfarne forskere kreves for å utføre von Frey testen og resultatene kan være subjektiv.
Automatisert gangart analyse systemet kan undersøke nevrologiske og nevromuskulære lidelser ved å måle ulike parametere gange i fritt flytte gnagere. I en rekke nerve skade dyr modeller, kan graden av nociception og antinociceptive effekten av flere behandlinger vurderes uten å legge en smerte stimulering11,12,13, 14. denne analysesystem finner statisk og dynamisk gangart parametere, som: pote utskriftsområde (området fullstendig paw print som kontakter med gulvet), pote intensitet (gjennomsnittlig intensiteten av kontakte pote området), skjærende lengden (den avstanden mellom etterfølgende plasseringer av det likt poten), holdning fase (varigheten av bakken kontakt for en enkelt bakben pote), trinnet sekvens (rekkefølgen som fire paws er plassert på gulvet), svinge (varigheten swing) og swing hastighet (beregnet fra stride lengde og svinge varighet og uttrykkes som piksler per sekund). Dette dokumentet demonstrerer bruken en analysesystem og gir en kort sammenligning av data med von Frey test med kronisk innsnevring skade (CCI) nevropatisk mus.

<table>
	<tbody>
		<tr>
			<td>0.9% Saline</td>
			<td>JW Pharmaceutical</td>
			<td>N/A</td>
			<td>Vehicle for drugs</td>
		</tr>
		<tr>
			<td>1ml syringe</td>
			<td>BD Plastipak</td>
			<td>300013</td>
			<td>Injecting device</td>
		</tr>
		<tr>
			<td>2, 2, 2-tribromoethanol (97% purity)</td>
			<td>Sigma</td>
			<td>T48402</td>
			<td>Anesthetic</td>
		</tr>
		<tr>
			<td>2-methyl-2-buthanol (99% purity)</td>
			<td>Sigma</td>
			<td>152463</td>
			<td>Solvent for 2, 2, 2-tribromoethanol</td>
		</tr>
		<tr>
			<td>Catwalk Automated gait analysis system</td>
			<td>Noldus</td>
			<td>N/A</td>
			<td>Automatic analysis software of aniaml gait</td>
		</tr>
		<tr>
			<td>Chromic catgut (4-0 thickness)</td>
			<td>AILEE</td>
			<td>C442</td>
			<td>Ligature to make chronic constriction injury on the sciatic nerve</td>
		</tr>
		<tr>
			<td>Gabapentin</td>
			<td>Sigma</td>
			<td>Y0001280</td>
			<td>Analgeisc, Used as a positive control drug in this study</td>
		</tr>
		<tr>
			<td>Graefe Forceps</td>
			<td>F.S.T</td>
			<td>11051-10</td>
			<td>Surgical instrument</td>
		</tr>
		<tr>
			<td>Heating Pad</td>
			<td>DAESHIN ELECTRONICS</td>
			<td>M-303AT</td>
			<td>Regulation of body temperature</td>
		</tr>
		<tr>
			<td>ICR Mouse</td>
			<td>Samtaco</td>
			<td>N/A</td>
			<td>Experimental animal</td>
		</tr>
		<tr>
			<td>Mersilk (3-0 thickness)</td>
			<td>ETHICON</td>
			<td>W598H</td>
			<td>Suture material for surgical closure of skin</td>
		</tr>
		<tr>
			<td>Micro-Mosquito</td>
			<td>F.S.T</td>
			<td>13010-12</td>
			<td>Surgical instrument</td>
		</tr>
		<tr>
			<td>Micro-scissors</td>
			<td>F.S.T</td>
			<td>14090-09</td>
			<td>Surgical instrument</td>
		</tr>
		<tr>
			<td>Needle holder</td>
			<td>F.S.T</td>
			<td>12002-12</td>
			<td>Surgical instrument</td>
		</tr>
		<tr>
			<td>Povidone Iodine</td>
			<td>Firson</td>
			<td>N/A</td>
			<td>Disinfectant to prevent infection after surgery</td>
		</tr>
		<tr>
			<td>Scalpel blade</td>
			<td>F.S.T</td>
			<td>10010-00 (#10)</td>
			<td>Surgical instrument to make an incision</td>
		</tr>
		<tr>
			<td>Scalpel handle</td>
			<td>F.S.T</td>
			<td>10003-12 (#3)</td>
			<td>Surgical instrument to make an incision</td>
		</tr>
		<tr>
			<td>Von-Frey filaments</td>
			<td>North Coast</td>
			<td>NC12775-99</td>
			<td>Measurement device to test sensory function for mechanical stimulation</td>
		</tr>
	</tbody>
</table>

automated gait analysis in mice with chronic constriction injury

Von Frey testen er en klassisk metode som har vært mye brukt til å undersøke funksjonen sensoriske nevropatisk smerte dyr. Men har det noen ulemper som subjektive data og kravet om en dyktig og erfaren eksperimentator. Hittil har en rekke endringer forbedret metoden von Frey, men fortsatt har noen begrensninger. Nylige rapporter har antydet at gait analyse produserer mer nøyaktig og objektive data fra nevropatisk dyrene. Denne protokollen demonstrerer hvordan å utføre automatiserte gait analyse for å fastslå graden av nevropatisk smerte i mus. Etter flere dager av acclimation, fikk mus lov å gå fritt på glassgulvet å belyse fotavtrykk. Deretter kvantifisering av fotavtrykk og gangart ble utført gjennom videoklipp med automatisk analyse av ulike gangavstand parametere, for eksempel område av paw print, swing tid, vinkelen på labben, etc.
Hovedformålet med denne studien er å beskrive metodikk automatisert gait analyse og kort sammenligne den med data fra klassisk sensoriske testen bruker von Frey filament.

Vi har utført von Frey testen og automatisert gait analyse i CCI mus til 10 dager etter CCI kirurgi. Gjentatte målinger av toveis variansanalyse (ANOVA) bestemmer den samlede effekten statistisk analyse og Dunnett's post-hoc analyse ble gjennomført for å fastslå den p-verdien blant de eksperimentelle gruppene.
Resultatene som vises i figur 1 angir time course of klassisk von Frey test-...

Watch this Scientific Journal Video about Automated Gait Analysis in Mice with Chronic Constriction Injury at JoVE.com

Automated Gait Analysis in Mice with Chronic Constriction Injury

Nøyaktig vurdering av smerte respons i en nevropatisk dyremodell er avgjørende å undersøke i Patofysiologien ved smerte sykdommer og utvikle nye smertestillende. Vi presenterer en følsom og objektive metode for å bestemme sensorisk funksjonen av gnagere hind pote av et automatisert gangart analysesystem.

Automatisert Gait analyse i mus med kronisk innsnevring skade

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 ...

The overall goal of this procedure is to analyze mechanical allodynia of mice with a chronic constriction injury using an automated gait analysis system. This method can help answer key questions in the pain research field by measuring sensory function after peripheral nerve injury in rodents. The main advantage of this technique is that the analysis system is quick and easy to use and it also automatically scores various gait parameters.For this study, house male ICR mice weighing 20 to 25 grams under standard conditions for at least one week. Select mice based on their appearance and behavior. Do not use mice that exhibit abnormal appearances or behavior such as hair loss, scarring and less movement.Using the selected mice, starting five days before the surgery collect automated gait data on a daily basis to have a baseline measurement. One day before the surgery perform the Von Frey test with a one gram filament on a hind paw to get a baseline response. Balance the responses between the injury group and the sham group.On each test day bring mice to the behavioral test room and acclimate the mice in their own home cage for at least 30 minutes before the test. Every other day after the surgery, measure mechanical allodynia using a Von Frey test. Put the mouse in a clean transparent acrylic box on a metal mesh floor and let it acclimate to the box for 30 minutes.Next, gently apply a one gram Von Frey filament to the plantar surface of the ipsilateral hind paw until the filament bends. Repeat this pattern of stimulation 10 times observing for paw withdrawals. Wait at least 10 seconds between stimulations.Record the number of withdrawals and score their frequency for analysis. Every other day, also perform the gait analysis test which must be done in the dark. Again, acclimate the mice to the test room in their home cage for at least 30 minutes.To set up the gait analysis device go to the software control. Under the setup tab set the walkway length to 30 centimeters, set the maximum run duration to five seconds and set the maximum run variation to 50%Next, select a registered camera from the setup tab of the program menu and then select open acquisition from the acquire tab. Following the status message, click the snap background button to take a background picture of the empty walkway.Now place a mouse onto the walkway and allow it to acclimate to the device for 10 minutes. The apparatus should always be washed with alcohol between mice. After the mouse has acclimated to the device, click the start acquisition button.The recording starts automatically according to the movement of the mouse. When the mouse clearly walks across the walkway, the program will automatically classify the compliant runs with a green icon. If the software fails to detect the footprints, a red icon will light up.After collecting at least five compliant runs select the classify runs option from the acquire tab of the program menu. Select which run should be analyzed and click the auto classify button. The results are automatically saved.To view the results click on view run statistics from the analyze menu. The Von Frey test and automated gait analysis were used to assess mice that underwent the CCI surgery on 10 consecutive days post-operatively. The frequency of paw withdrawal increased two days after the surgery and was sustained for 10 days.This effect could be significantly reduced by daily post-operative injections of gabapentin. Using the gait analysis software the size of the left and right hind paw prints could be compared. Normally the areas are the same which is indicated by a 50%value.The CCI surgery had a significant effect on paw print and that effect could be significantly reduced using gabapentin. The gait analysis software is also able to pick up on the change of single stance durations when the mouse is weight-bearing on just one hind paw. This is calculated and shown as a percent change.This metric shows the same effect for the CCI surgery and gabapentin treatments. After watching this video you should have a good understanding of how to operate and analyze automated gait analysis systems to investigate the pathophysiology of neuropathic pain in animal models like chronic constriction injury.

Research

JoVE Journal

Neuroscience

Automated Sholl Analysis of Digitized Neuronal Morphology at Multiple Scales

Neuronal morphology plays a significant role in determining how neurons function and communicate1-3.  Specifically, it affects the ability of neurons to ...

Spectral Confocal Imaging of Fluorescently tagged Nicotinic Receptors in Knock-in Mice with Chronic Nicotine Administration

Spectral confocal Imaging av fluorescently tagget nikotinreseptorene i Knock-in Mus med kronisk Nikotin Administrasjon

Ligand-gated ion channels in the central nervous system (CNS) are implicated in numerous conditions with serious medical and social consequences.  For ...

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

Sagittal flyet Kinematisk Gait analyse i C57BL/6 mus utsatt for MOG35-55 indusert eksperimentelle Autoimmune immunsviktvirus

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

Chronic Implantation of Whole-cortical Electrocorticographic Array in the Common Marmoset

Kronisk implantering av hele-kortikale Electrocorticographic matrise i den vanlige Marmoset

Electrocorticography (ECoG) allows the monitoring of electrical field potentials from the cerebral cortex with high spatiotemporal resolution. Recent ...

Automated Gait Analysis to Assess Functional Recovery in Rodents with Peripheral Nerve or Spinal Cord Contusion Injury

Automatisert gangart analyse for å vurdere funksjonell utvinning hos gnagere med perifer nerve eller ryggmargskontusjonsskade

Peripheral and central nerve injuries are mostly studied in rodents, especially rats, given the fact that these animal models are both cost-effective and ...

Comprehensive Understanding of Inactivity-Induced Gait Alteration in Rodents

Omfattende forståelse av inaktivitetsindusert gangendring hos gnagere

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

Electromagnetic Controlled Closed-Head Model of Mild Traumatic Brain Injury in Mice

Elektromagnetisk kontrollert lukket hodemodell av mild traumatisk hjerneskade hos mus

Highly reproducible animal models of traumatic brain injury (TBI), with well-defined pathologies, are needed for testing therapeutic interventions and ...

En kontusjonsenhet for ryggmargsskade for nøyaktige musemodeller

Automated Impactor for Contusive Spinal Cord Injury Model in Mice

Forfatter i søkelyset: Innsikt i innovasjoner innen ryggmargsskadeforskning

Automatisert impactor for kontusiv ryggmargsskademodell hos mus

Spinal cord injury (SCI) due to traumatic injuries such as car accidents and falls is associated with permanent spinal cord dysfunction. Creation of ...