Name: urokinase-type plasminogen activator-induced mouse back pain model
Uploaded: 2023-09-01T12:00:00
Description: Watch this Scientific Journal Video about Urokinase-type Plasminogen Activator-induced Mouse Back Pain Model at JoVE.com

Tilskudsfinansiering blev leveret af NIH HEAL UG3 NS123958. Boligfaciliteterne blev inspiceret og akkrediteret af AAALAC. Dyrene blev anbragt i Animal Resources Center (ARC) opstaldningsfaciliteter, der blev vedligeholdt af laboratoriepersonalet og Division of Laboratory and Animal Resources (DLAR) personale. Procedurerne for adfærdstest er standardmetoder på området som godkendt af American Pain Society og International Association for Study of Pain. Metoden til eutanasi er i overensstemmelse med anbefalingerne fra panelet om eutanasi fra American Veterinary Medical Association.

Alle beskrevne dyreforsøg er i overensstemmelse med NIH-vejledningen til pleje og brug af forsøgsdyr. Undersøgelser blev godkendt af det lokale institutionelle pleje- og brugsudvalg (IACUC # 23-201364-HSC) ved University of New Mexico Health Sciences Center. Alle undersøgelser er i overensstemmelse med politikker i regi af en OLAW Assurance of Compliance (A3002-01) om anvendelse af dyr i forskning, som beskrevet i del III. II. Forsikringer og certificeringer. Dyrene er anbragt i Animal Resources Center (ARC) opstaldn...

<ol>
	<li>O'Sullivan, P. 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=Back+to+basics:+10+facts+every+person+should+know+about+back+pain.">Back to basics: 10 facts every person should know about back pain.</a> British Journal of Sports Medicine. 54 (12), 698-699 (2020).</li><li>Nauta, H. J., McIlwrath, S. L., Westlund, K. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Punctate+midline+myelotomy+reduces+pain+responses+in+a+rat+model+of+lumbar+spine+pain:+evidence+that+the+postsynaptic+dorsal+column+pathway+conveys+pain+from+the+axial+spine.">Punctate midline myelotomy reduces pain responses in a rat model of lumbar spine pain: evidence that the postsynaptic dorsal column pathway conveys pain from the axial spine.</a> Cureus. 10 (3), 2371 (2018).</li><li>Shuang, 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=Establishment+of+a+rat+model+of+lumbar+facet+joint+osteoarthritis+using+intraarticular+injection+of+urinary+plasminogen+activator.">Establishment of a rat model of lumbar facet joint osteoarthritis using intraarticular injection of urinary plasminogen activator.</a> Scientific Reports. 5 (1), 9828 (2015).</li><li>Reed, N. R., 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=Somatosensory+behavioral+alterations+in+a+NGF-induced+persistent+low+back+pain+model.">Somatosensory behavioral alterations in a NGF-induced persistent low back pain model.</a> Behavioural Brain Research. 418, 113617 (2022).</li><li>Suh, H. R., Cho, H. -. Y., Han, H. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+a+novel+model+of+intervertebral+disc+degeneration+by+the+intradiscal+application+of+monosodium+iodoacetate+(MIA)+in+rat.">Development of a novel model of intervertebral disc degeneration by the intradiscal application of monosodium iodoacetate (MIA) in rat.</a> The Spine Journal. 22 (1), 183-192 (2022).</li><li>Kim, H., Hong, J. Y., Lee, J., Jeon, W. -. J., Ha, I. -. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=IL-1&#946;+promotes+disc+degeneration+and+inflammation+through+direct+injection+of+intervertebral+disc+in+a+rat+lumbar+disc+herniation+model.">IL-1&#946; promotes disc degeneration and inflammation through direct injection of intervertebral disc in a rat lumbar disc herniation model.</a> The Spine Journal. 21 (6), 1031-1041 (2021).</li><li>Millecamps, M., Tajerian, M., Sage, E. H., Stone, L. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+signs+of+chronic+back+pain+in+the+SPARC-null+mouse.">Behavioral signs of chronic back pain in the SPARC-null mouse.</a> Spine. 36 (2), 95-102 (2011).</li><li>La Porta, C., Tappe-Theodor, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+impact+of+psychological+and+psychophysical+stress+on+low+back+pain+in+mice.">Differential impact of psychological and psychophysical stress on low back pain in mice.</a> Pain. 161 (7), 1442-1458 (2020).</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> Journal of Neuroscience Methods. 53 (1), 55-63 (1994).</li><li>Takao, K., Miyakawa, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Light/dark+transition+test+for+mice.">Light/dark transition test for mice.</a> Journal of Visualized Experiments JoVE. (1), e104 (2006).</li><li>Deuis, J. R., Dvorakova, L. S., Vetter, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methods+used+to+evaluate+pain+behaviors+in+rodents.">Methods used to evaluate pain behaviors in rodents.</a> Frontiers in Molecular Neuroscience. 10, 284 (2017).</li><li>David, D. 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=Neurogenesis-dependent+and+-independent+effects+of+fluoxetine+in+an+animal+model+of+anxiety/depression.">Neurogenesis-dependent and -independent effects of fluoxetine in an animal model of anxiety/depression.</a> Neuron. 62 (4), 479-493 (2009).</li><li>Yalcin, I., 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=A+time-dependent+history+of+mood+disorders+in+a+murine+model+of+neuropathic+pain.">A time-dependent history of mood disorders in a murine model of neuropathic pain.</a> Biological Psychiatry. 70 (10), 946-953 (2011).</li><li>Madathil, S. 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=Astrocyte-specific+overexpression+of+insulin-like+growth+factor-1+protects+hippocampal+neurons+and+reduces+behavioral+deficits+following+traumatic+brain+injury+in+mice.">Astrocyte-specific overexpression of insulin-like growth factor-1 protects hippocampal neurons and reduces behavioral deficits following traumatic brain injury in mice.</a> PloS One. 8 (6), e67204 (2013).</li><li>Sugimoto, H., Kawakami, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Low-cost+protocol+of+footprint+analysis+and+hanging+box+test+for+mice+applied+the+chronic+restraint+stress.">Low-cost protocol of footprint analysis and hanging box test for mice applied the chronic restraint stress.</a> Journal of Visualized Experiments: JoVE. (143), e59027 (2019).</li><li>Hassan, 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=Identifying+chronic+low+back+pain+phenotypic+domains+and+characteristics+accounting+for+individual+variation:+a+systematic+review.">Identifying chronic low back pain phenotypic domains and characteristics accounting for individual variation: a systematic review.</a> Pain. , (2023).</li></ol>

Denne model af kroniske rygsmerter er enkel at inducere, og overfølsomhed etableret inden for 1 uge kan vare i op til (og muligvis ud over) 8 uger. Dette giver mulighed for nøjagtig undersøgelse af kronisk smertetilstand i modsætning til andre akutte modeller, der kun varer i en uge eller to. Mens vi viser modellen i mus, kan den uPA-inducerede CBP-model også etableres i rotter2. En fordel ved modellen er, at det langvarige tidsforløb fremkalder udviklingen af angst- og depressionslignende a...

Lændesmerter er en af de mest almindelige årsager til handicap med 1 ud af 5 mennesker, der lider over hele verden1. På trods af disse bestræbelser anvendes få pålidelige dyremodeller af rygsmerter populært i dyreforsøg på smerteområdet, især hos mus. Tidligere modeller har næsten udelukkende gjort brug af rotter til induktion af kroniske rygsmerter (CBP), såsom dem, der induceres ved injektion af urinplasminogenaktivator (uPA) i lændehvirvelsøjlen 2,3, injektion af nervevækstfaktor (NGF) i trunkmuskulatur4 eller mononatriumiodoacetat (MIA)5 eller interleukin-1beta6 injektion i den intravertebrale disk. Selvfølgelig foretrækkes rotter til disse modeller, hovedsageligt på grund af deres større størrelse og lette adgang til injektion af inflammatoriske midler.
For at være klar findes der musemodeller af rygsmerter, såsom SPARC-null-musemodellen for intervertebral diskdegeneration, der blev brugt i mange år7, men disse er dyrere og tidskrævende at etablere end injektionsbaserede modeller. En nylig museundersøgelse etablerede en model for lændesmerter ved at kombinere NGF-injektion i lændemuskler med lodret kronisk fastholdelsesstress8. I den følgende protokol har vi tilpasset den uPA-inducerede CBP-model fra rotter til mus2. Overfølsomhed etableres inden for 1 uge og vedvarer op til 6-8 uger. Derudover fastslår vi, at mus udvikler angst- og depressionslignende adfærd. I betragtning af forekomsten af rygsmerter og den mere almindelige brug af mus i molekylær smerteforskning, er denne holdbare model let etableret til brug i udviklingen af nye behandlingsstrategier til lindring af rygsmerter.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Animals and Consumables</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>70% ethanol</td>
			<td>Local Source</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>BALB/c mice</td>
			<td>Envigo</td>
			<td></td>
			<td>20-25 g</td>
		</tr>
		<tr>
			<td>Cotton balls</td>
			<td>Fisher Scientific</td>
			<td>19-090-702</td>
			<td></td>
		</tr>
		<tr>
			<td>Cotton-tipped applicators</td>
			<td>Fisher Scientific</td>
			<td>19-062-616</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane inhalant anesthetic</td>
			<td>MedVet</td>
			<td>RXISO-250</td>
			<td></td>
		</tr>
		<tr>
			<td>Labeling tape</td>
			<td>Fisher Scientific</td>
			<td>NGFP7002</td>
			<td></td>
		</tr>
		<tr>
			<td>Nitrile exam gloves</td>
			<td>Fisher Scientific</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Oxygen tank</td>
			<td>Local Source</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical drape, Steri-Drape Utility Sheet, Absorbent Prevention</td>
			<td>VWR</td>
			<td>76246-788</td>
			<td>cut into 15 x 15 cm pieces</td>
		</tr>
		<tr>
			<td>Tygon tubing with 3 mm inner diameter</td>
			<td>Grainger</td>
			<td>22XH87</td>
			<td></td>
		</tr>
		<tr>
			<td>Equipment</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>#11 carbon steel scalpel blades</td>
			<td>VWR</td>
			<td>21909-612</td>
			<td></td>
		</tr>
		<tr>
			<td>Anesthesia induction chamber&#160;</td>
			<td>Summit Medical Equipment Company</td>
			<td>AS-01-0530-LG</td>
			<td></td>
		</tr>
		<tr>
			<td>Autoclave</td>
			<td>Local Unit</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Biology Dumont #5 forceps</td>
			<td>Fine Science Tools</td>
			<td>11252-30</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass bead sterilizer Germinator 500</td>
			<td>VWR</td>
			<td>102095-946</td>
			<td></td>
		</tr>
		<tr>
			<td>IITC Life Sciences Series 8 Model PE34 Hot/Cold Plate Analgesia Meter</td>
			<td>IITC</td>
			<td>PE34</td>
			<td></td>
		</tr>
		<tr>
			<td>Integra Miltex cotton &#38; dressing pliers&#160;</td>
			<td>Safco Dental Supply</td>
			<td>66-317</td>
			<td></td>
		</tr>
		<tr>
			<td>OPTIKA CL31 double arm LED illuminator&#160;</td>
			<td>New York Microscope Company</td>
			<td>OPCL-31</td>
			<td></td>
		</tr>
		<tr>
			<td>Plantar Test System with InfraRed Emitter, i. e. Hargreaves Apparatus</td>
			<td>Ugo Basile</td>
			<td>37370-001 and 37370-002</td>
			<td></td>
		</tr>
		<tr>
			<td>Scalpel Handle No. 3</td>
			<td>VWR</td>
			<td>25607-947</td>
			<td></td>
		</tr>
		<tr>
			<td>Small animal heating pad&#160;</td>
			<td>Valley Vet Supply</td>
			<td>47375</td>
			<td></td>
		</tr>
		<tr>
			<td>Student Vannas spring scissors, straight blade&#160;</td>
			<td>Fine Science Tools</td>
			<td>91500-09</td>
			<td></td>
		</tr>
		<tr>
			<td>Table top animal research portable anesthesia workstation &#8220;PAM&#8221;</td>
			<td>Patterson Scientific</td>
			<td>AS-01-0007</td>
			<td></td>
		</tr>
		<tr>
			<td>Von Frey Filaments</td>
			<td>Ugo Basile</td>
			<td>37450-275</td>
			<td></td>
		</tr>
	</tbody>
</table>

urokinase-type plasminogen activator-induced mouse back pain model

En model af vedvarende lændesmerter kan induceres hos mus med den enkle metode, der er beskrevet heri. Trinvise metoder til enkel, hurtig induktion af en vedvarende rygsmertemodel hos mus tilvejebringes her ved anvendelse af en injektion af plasminogenaktivator af urokinase-typen (urokinase), en serinprotease, der findes hos mennesker og andre dyr. Metoden til induktion af vedvarende lændesmerter hos mus involverer en simpel injektion af urokinase langs ledbåndsindsættelsesområdet i lændehvirvelsøjlen. Det urokinase inflammatoriske middel aktiverer plasminogen til plasmin. Typisk kan modellen induceres inden for 10 minutter, og overfølsomheden vedvarer i mindst 8 uger.
Overfølsomhed, gangforstyrrelser og andre standard angst- og depressionslignende foranstaltninger kan testes i den vedvarende model. Rygsmerter er den mest udbredte type smerte. For at forbedre bevidstheden om rygsmerter udnævnte International Association for the Study of Pain (IASP) 2021 til "Global Year about Back Pain" og 2022 til "Global Year for Translating Pain Knowledge to Practice." En begrænsning af den terapeutiske udvikling af smerteterapi er manglen på egnede modeller til test af vedvarende og kroniske smerter. Funktionerne i denne model er egnede til at teste potentielle behandlinger rettet mod reduktion af rygsmerter og dets hjælpeegenskaber, hvilket bidrager til IASP's navngivning af 2022 som det globale år for oversættelse af smerteviden til praksis.

Nociceptiv-relateret adfærdstest og dataanalyse Fremkaldte foranstaltninger Overfølsomhed på fodpuden udvikler sig inden for en dag efter urokinaseinjektion. Inden for 1 uge sænkes tilbagetrækningstærsklen betydeligt og fortsætter indtil eutanasi; Dette er vist gennem postkirurgisk uge 4 (figur 4A). Pote tilbagetrækning latens analyseres ved hjælp af von Frey op-ned metode9 og Hargreaves test. I det afbildede eksempel v...

Watch this Scientific Journal Video about Urokinase-type Plasminogen Activator-induced Mouse Back Pain Model at JoVE.com

Urokinase-type Plasminogen Activator-induced Mouse Back Pain Model

Metoder til enkel, hurtig induktion af en rygsmertemodel hos mus tilvejebringes her ved anvendelse af en intraligamentinjektion af urinplasminogenaktivator.

Urokinase-type plasminogen aktivator-induceret mus rygsmerter model

A model of persisting lower back pain can be induced in mice with the simple methodology described herein. Step-by-step methods for simple, rapid ...

Denne protokol skitserer en ikke-invasiv metode til at fremkalde kroniske rygsmerter hos mus i op til 10 uger, som kan udføres hurtigt og pålideligt med minimal træning. Det sværeste aspekt af denne teknik er at genkende, hvordan indsprøjtning af ledbånd føles versus injektion af muskler. Følelse af modstand ved kontakt indikerer ledbåndsinjektion.Ingen modstand betyder, at nålen kan være i musklen eller bughulen. Til at begynde med skal du tage en steril flad overflade for at placere musen. Placer derefter en varmepudegenvindingsstation ved siden af operationsopsætningen for straks at overføre mus efter injektion.Forbered derefter urokinase og en Hamilton-sprøjte. Tegn opløsningen på forhånd for at minimere den tid, musen bruger under anæstesi. Efter bedøvelse af musen skal du bruge to fingre til at lokalisere lændehvirvelsøjlesegmenterne ved L2 og L3 under brystkassen og kontrollere injektionsstedet.Placer spidsen af Hamilton-sprøjten ved siden af rygsøjlen. Placer sprøjten i en vinkel på ca. 45 grader ind i ledbåndet umiddelbart ved siden af knoglen. Indsæt nålespidsen forsigtigt, men fast, i ledbåndet, og injicer langsomt, indtil alle fem mikroliter er injiceret.Fjern nålen forsigtigt og langsomt. Placer derefter musen i varmegenvindingsstationen, indtil den vågner og bliver mobil. Kontroller musene i en time efter operationen for at sikre, at al normal motorisk funktion fortsætter.Efter operationen skal du kontrollere musenes vægt og injektionssted dagligt i en uge for at forhindre infektion eller komplikationer. Før testning flyttes hvert dyr til en separat klar kabine på testbordet med en skærmtop. Stimuler derefter fodpuden med 3.61 Von Frey-filamentet, der fremkalder fire gram kraft.En fodtilbagetrækning på tre til fem stimuli betragtes som et positivt svar. Anvend nu det næste svagere filament i serien, indtil dyret holder op med at reagere på den mekaniske stimulering. Brug det højere filament, indtil der fremkaldes et svar, og skift derefter tilbage til det nederste filament.Ved hjælp af en kurvetilpasningsalgoritme beregnes den mekaniske tilbagetrækningstærskel, som er den minimale mængde kraft, der er nødvendig for at fremkalde et svar 50% af tiden. For at udføre Hargreaves-testen skal du placere musene i kabiner på en glasoverflade, der opvarmes nedenfra af en infrarød sender. Mål tiden fra påføring af infrarød lysstimulus på musens bagpote til tilbagetrækning som fodlatenstid.Til koldpladetesten placeres musene på koldpladeapparatet, der er afkølet til minus ni grader Celsius. Registrer den tid, musen tager at løfte foden efter at være blevet placeret på apparatet som latenstid for at trække foden tilbage. For at udføre angsttesten skal du placere hvert dyr i testboksen til stedpræference med en passage mellem to kamre.Et apparatkammer skal være stærkt oplyst, mens den modsatte side skal forblive mørk. Brug en computer til at overvåge dyrets lyse og mørke belægningstider og overgange i løbet af en 10-minutters test. Placer derefter modelgnavere eller naive dyr på nullabyrinten, eller brug to-kammer lys eller mørk stedpræferencetest.Vurder den tid, der bruges i lukkede og åbne områder af labyrinten ved hjælp af et stopur. Til Succharose Splash Depression Test sprøjtes en 10 til 30% saccharoseopløsning på dorsalkoden og scorer hyppigheden, varigheden og latenstiden for pleje i 10 minutter. For at udføre nye objekttest skal du akklimatisere mus individuelt til et klart plastbur med en åben top i en time.Tilføj to identiske legetøjsminifigurer i hvert sit hjørne af buret i fem minutter. Akklimatisere dyrene på testdagen i en time, og placer de to identiske minifigurer i de samme positioner i buret i fem minutter, før de returneres til hjemmeburet. Udskift derefter en af de originale figurer med et tydeligt andet romanobjekt.Efter fire timer skal du returnere musene til testburet og registrere den tid, du bruger på at udforske objekterne. Til vurdering af motorfunktionen konstrueres en tunnel af et papirhåndklæderør skåret i længderetningen på den ene kant. Spred tunnelen ud på et rent stykke printerpapir.Hold musene forsigtigt, indtil de er rolige. Påfør giftfri indisk blæk på en mus' poter ved at placere dem i en blækbrønd, stempelpude eller ved hjælp af en vatpind. Slip musene løs ved tunnelindgangen.Lad dem løbe gennem tunnelen og fange dem i slutningen. Scor poteaftryk baseret på skridtlængde, skridtbredde og tåspredning. Mekaniske og termiske overfølsomhedsanalyser viste, at mus med kroniske rygsmerter havde signifikant øget mekanisk, varme- og kuldefølsomhed sammenlignet med de naive kontroller.Angstanalyse viste, at den samlede tid brugt i lyskammeret i den lyse og mørke boks og antallet af opdrætshændelser var signifikant lavere hos mus med kroniske rygsmerter, hvilket indikerer angst. Under Succharose Splash Test udviste mus med kroniske rygsmerter lavere plejetid og antallet af pleje. Imidlertid blev mængden af tid, før plejen starter, signifikant øget.Skridtlængden mellem mus med kroniske rygsmerter og naive mus var signifikant forskellig. Den vigtigste ting at huske, når du forsøger denne procedure, er, at du sigter mod ledbåndene omkring rygsøjlen, ikke musklen. Den kroniske rygsmerter model giver et pålideligt værktøj til at vurdere en række nye forskning, farmakologiske og andre potentielle behandlinger.

urokinase-type-plasminogen-activator-induced-mouse-back-pain-model

Research

JoVE Journal

Behavior

Assessing Functional Performance in the Mdx Mouse Model

Duchenne muscular dystrophy (DMD) is a severe and progressive muscle wasting disorder for which no cure is available. Nevertheless, several potential pharmaceutical compounds and gene therapy approaches have progressed into clinical trials. With improvement in muscle function being the most important end point in these trials, a lot of emphasis has been placed on setting up reliable, reproducible, and easy to perform functional tests to pre clinically assess muscle function, strength, condition, and coordination in the mdx mouse model for DMD. Both invasive and noninvasive tests are available. Tests that do not exacerbate the disease can be used to determine the natural history of the disease and the effects of therapeutic interventions (e.g. forelimb grip strength test, two different hanging tests using either a wire or a grid and rotarod running). Alternatively, forced treadmill running can be used to enhance disease progression and/or assess protective effects of therapeutic interventions on disease pathology. We here describe how to perform these most commonly used functional tests in a reliable and reproducible manner. Using these protocols based on standard operating procedures enables comparison of data between different laboratories.

Assessing Functional Performance in the Mdx Mouse Model

The primary outcome measure in clinical trials for neuromuscular disorders is generally improved muscle function. Therefore, assessing the effect of potential therapeutic compounds on muscle performance pre clinically in mouse models is of great importance. We here describe several functional tests to address this.

Vurdere Funktionel Performance i<em&gt; Mdx</em&gt; Mus Model

Duchenne muscular dystrophy (DMD) is a severe and progressive muscle wasting disorder for which no cure is available. Nevertheless, several potential ...

Determining Pain Detection and Tolerance Thresholds Using an Integrated, Multi-Modal Pain Task Battery

Human pain models are useful in the assessing the analgesic effect of drugs, providing information about a drug&#39;s pharmacology and identify potentially suitable therapeutic populations. The need to use a comprehensive battery of pain models is highlighted by studies whereby only a single pain model, thought to relate to the clinical situation, demonstrates lack of efficacy. No single experimental model can mimic the complex nature of clinical pain. The integrated, multi-modal pain task&#160;battery presented here encompasses the electrical stimulation task, pressure stimulation task, cold pressor task, the UVB inflammatory model which includes a thermal task and a paradigm for inhibitory conditioned pain modulation. These human pain models have been tested for predicative validity and reliability both in their own right and in combination, and can be used repeatedly, quickly, in short succession, with minimum burden for the subject and with a modest quantity of equipment. This allows a drug to be fully characterized and profiled for analgesic effect which is especially useful for drugs with a novel or untested mechanism of action.

Human pain models are valuable tools used to assess the analgesic potential of novel compounds and predict their clinical efficacy, especially when used in an integrated manner. Although implementation of these models is complex, with proper execution, the pain models described in this protocol can provide predictive and reliable results.

Bestemmelse Smerter Detection og tolerancetærskler Ved hjælp af en integreret, Multi-Modal Pain Task Batteri

Human pain models are useful in the assessing the analgesic effect of drugs, providing information about a drug&#39;s pharmacology and identify ...

Treating Low Back Pain in Failed Back Surgery Patients with Multicolumn-lead Spinal Cord Stimulation

Failed back surgery syndrome (FBSS) refers to persistent, chronic pain following spinal surgery. Spinal cord stimulation with dorsal epidural leads can be used to treat back and leg pain in FBSS patients. This paper presents a detailed protocol for using spinal cord stimulation with surgical leads in FBSS patients. In our department, with the patient under general anesthesia, we place the lead in the epidural space by means of a small laminectomy at the 10th thoracic level. Placement of the lead is followed by a 1 month trial period with an externalized lead. If pain relief is greater than 50% at the end of this 1 month stimulation trial (required by Belgian reimbursement criteria), an internal pulse generator is then placed under the skin and connected to the lead in a second surgical procedure. We have demonstrated that using this technique in rigorously selected FBSS patients can significantly improve back pain, leg pain, patient activity, and quality of life for a sustained period of time.

This paper presents a method that uses spinal cord stimulation with a multicolumn lead to treat neuropathic low back pain in failed back surgery patients.

Behandling af lændesmerter i fejlslagen rygkirurgi patienter med Flersøjle-bly rygmarven Stimulation

Failed back surgery syndrome (FBSS) refers to persistent, chronic pain following spinal surgery. Spinal cord stimulation with dorsal epidural leads can be ...

Measuring Active and Passive Tameness Separately in Mice

Domesticated animals such as dogs and laboratory mice show a high level of tameness, which is important for humans to handle them easily. Tameness has two behavioral components: a reluctance to avoid humans (passive tameness) and a motivation to approach humans (active tameness). To quantify these components in mice, we previously developed behavioral tests for active tameness, passive tameness, and the willingness to stay on a human hand, each designed to be completed within 3 min. The data obtained were used for selective breeding, with a large number of mice analyzed per generation. The active tameness test measures the movement of the mouse toward a human hand and the contact it engages in. The passive tameness test measures the duration of time that a mouse tolerates human touch. In the stay-on-hand test, a mouse is placed on a human hand and touched slowly using the thumb of that hand; the duration of time that the animal remains on the hand is measured. Here, we describe the test set-up and apparatus, explain the procedures, and discuss the appropriate data analysis. Finally, we explain how to interpret the results.

Tameness in animals includes a reduction of avoidance responses to humans (passive tameness) and an increase in active approaches to humans (active tameness). Here, we describe detailed protocols for three behavioral tests (active tameness, passive tameness, and stay-on-hand tests) to separately measure the active and passive tameness of mice.

Måling af aktive og Passive Tameness separat i mus

Domesticated animals such as dogs and laboratory mice show a high level of tameness, which is important for humans to handle them easily. Tameness has two ...

Chronic Post-Ischemia Pain Model for Complex Regional Pain Syndrome Type-I in Rats

Complex regional pain syndrome type-I (CRPS-I) is a neurological disease that causes severe pain among patients and remains an unresolved medical condition. However, the underlying mechanisms of CRPS-I have yet to be revealed. It is known that ischemia/reperfusion is one of the leading factors that causes CRPS-I. By means of prolonged ischemia and reperfusion of the hind limb, the rat chronic post-ischemia pain (CPIP) model has been established to mimic CRPS-I. The CPIP model has become a well-recognized animal model for studying the mechanisms of CRPS-I. This protocol describes the detailed procedures involved in the establishment of the rat model of CPIP, including anesthesia, followed by ischemia/reperfusion of the hind limb. Characteristics of the rat CPIP model are further evaluated by measuring the mechanical and thermal hypersensitivities of the hind limb as well as the nocifensive responses to acute capsaicin injection. The rat CPIP model exhibits several CRPS-I-like manifestations, including hind limb edema and hyperemia in the early stage after establishment, persistent thermal and mechanical hypersensitivities, and increased nocifensive responses to acute capsaicin injection. These characteristics render it a suitable animal model for further investigation of the mechanisms involved in CRPS-I.

Provided here is a protocol that details steps to establish an animal model of chronic post-ischemia pain (CPIP). This is a well-recognized model mimicking human complex regional pain syndrome type-I. Mechanical and thermal hypersensitivities are further evaluated, as well as capsaicin-induced nocifensive behaviors observed in the CPIP rat model.

Kronisk post-iskæmi smerte model for komplekse regionale smerte syndrom type-i i rotter

Complex regional pain syndrome type-I (CRPS-I) is a neurological disease that causes severe pain among patients and remains an unresolved medical ...

Investigating Pain-Related Avoidance Behavior using a Robotic Arm-Reaching Paradigm

Avoidance behavior is a key contributor to the transition from acute pain to chronic pain disability. Yet, there has been a lack of ecologically valid paradigms to experimentally investigate pain-related avoidance. To fill this gap, we developed a paradigm (the robotic arm-reaching paradigm) to investigate the mechanisms underlying the development of pain-related avoidance behavior. Existing avoidance paradigms (mostly in the context of anxiety research) have often operationalized avoidance as an experimenter-instructed, low-cost response, superimposed on stimuli associated with threat during a Pavlovian fear conditioning procedure. In contrast, the current method offers increased ecological validity in terms of instrumental learning (acquisition) of avoidance, and by adding a cost to the avoidance response. In the paradigm, participants perform arm-reaching movements from a starting point to a target using a robotic arm, and freely choose between three different movement trajectories to do so. The movement trajectories differ in probability of being paired with a painful electrocutaneous stimulus, and in required effort in terms of deviation and resistance. Specifically, the painful stimulus can be (partly) avoided at the cost of performing movements requiring increased effort. Avoidance behavior is operationalized as the maximal deviation from the shortest trajectory on each trial. In addition to explaining how the new paradigm can help understand the acquisition of avoidance, we describe adaptations of the robotic arm-reaching paradigm for (1) examining the spread of avoidance to other stimuli (generalization), (2) modeling clinical treatment in the lab (extinction of avoidance using response prevention), as well as (3) modeling relapse, and return of avoidance following extinction (spontaneous recovery). Given the increased ecological validity, and numerous possibilities for extensions and/or adaptations, the robotic arm-reaching paradigm offers a promising tool to facilitate the investigation of avoidance behavior and to further our understanding of its underlying processes.

Avoidance is central to chronic pain disability, yet adequate paradigms for examining pain-related avoidance are lacking. Therefore, we developed a paradigm that allows investigating how pain-related avoidance behavior is learned (acquisition), spreads to other stimuli (generalization), can be mitigated (extinction), and how it may subsequently re-emerge (spontaneous recovery).

Undersøgelse af smerterelateret undgåelsesadfærd ved hjælp af et robotarmsnået paradigme

Avoidance behavior is a key contributor to the transition from acute pain to chronic pain disability. Yet, there has been a lack of ecologically valid ...

Back Mechanical Sensitivity Assessment in the Rat for Mechanistic Investigation of Chronic Back Pain

Low back pain is the leading cause of disability worldwide, with dramatic personal, economic, and social consequences. To develop novel therapeutics, animal models are needed to examine the mechanisms and effectiveness of novel therapies from a translational perspective. Several rodent models of back pain are used in current investigations. Surprisingly, however, no standardized behavioral test was validated to assess mechanical sensitivity in back pain models. This is critical to confirm that animals with presumed back pain present local hypersensitivity to nociceptive stimuli, and to monitor sensitivity during interventions designed to relieve back pain. The objective of this study is to lay down a simple and accessible test to assess mechanical sensitivity in the back of rats. A test cage was fabricated specifically for this method; length x width x height: 50 x 20 x 7 cm, having a stainless-steel mesh on the top. This test cage allows the application of mechanical stimuli to the back. To perform the test, the back of the animal is shaved in the region of interest, and the test area is marked to repeat the test on different days, as needed. The mechanical threshold is determined with Von Frey filaments applied to the paraspinal muscles, utilizing the up-down method described previously. The positive responses include (1) muscle twitching, (2) arching (back extension), (3) rotation of the neck (4) scratching or licking the back, and (5) escaping. This behavioral test (Back Mechanical Sensitivity (BMS) test) is useful for mechanistic research with rodent models of back pain for the development of therapeutic interventions for the prevention and management of back pain.

To develop novel therapeutic interventions for the prevention and management of back pain, animal models are required to examine the mechanisms and effectiveness of these therapies from a translational perspective. The present protocol describes the BMS test, a standardized method to assess back mechanical sensitivity in the rat.

Vurdering af rygmekanisk følsomhed hos rotter til mekanistisk undersøgelse af kroniske rygsmerter

Low back pain is the leading cause of disability worldwide, with dramatic personal, economic, and social consequences. To develop novel therapeutics, ...

Elevated Plus Maze Test in an Angelman Syndrome Mouse Model

To begin, place the plus shaped maze on the testing platform just below the camera. Using the potentiometer on the wall, adjust the light intensity to 70 luxe at its center with the help of a luxometer with its sensor placed at the center of the maze during the adjustment. Open the software by double clicking on the viewer software icon and load the configuration for Elevated Plus Maze or EPM testing by clicking on the icon on the upper left side of the configuration tab.Enter the animal information into the corresponding fields of the experiment tab including animal id, genotype, gender, and experiment information. Verify that the zone's position, open arms, and closed arms are correctly configured. Using a visual control and computer mouse, ensure that the virtual outline zones match the corresponding EPM zones.Place the mouse cursor on the arrow icon on the upper left side of the acquisition tab. Remove the animal from the home cage and place it gently in the center of the EPM. Start the experiment by left clicking on the computer mouse.After five minutes, save the recorded data by clicking okay. Name the file and click save. Export the results to a csv file for offline analysis by clicking on the icon on the left vertical panel of the data analysis tab.In the EPM test, none of the parameters related to anxiety-like behavior were altered in maternal heterozygous Ube3A mutants.

Forhøjet plus labyrinttest i en Angelman syndrom musemodel

Open Field Test in an Angelman Syndrome Mouse Model

Place the four open field, or OF, test boxes on the testing platform below the camera. Using the potentiometer on the wall, adjust the light intensity to 200 lux at the center of each OF test with the help of a luxometer with its sensor placed at the center of each box during the adjustment. Open the software by double clicking on the viewer software icon and load the configuration for EF testing by clicking on the icon on the upper left side of the configuration tab.Enter the animal information into the corresponding fields of the experiment tab. Verify that the zone's position matches the OF test boxes and adjust them if needed. Using a visual control and computer mouse, ensure that the virtual outline center and periphery zones match the corresponding OF test zones.Place the mouse cursor on the arrow icon on the upper left side of the acquisition tab. Remove four animals from the home cage and place them in the corner of each test box. Stop the experiment by left clicking on the computer mouse.In the OF test, the maternal heterozygous Ube3a mutant animals were significantly hypoactive as reflected by a shorter traversed distance, lower average speed, and longer resting time. However, none of the parameters related to anxiety-like behavior were altered in Ube3a mutants.

Åben felttest i en Angelman syndrom musemodel

Tail Suspension Test in an Angelman Syndrome Mouse Model

Connect the tail suspension cage to the computer using a USB cable. Insert the USB dongle into the computer and start the software by double clicking on the BioTSD software icon. In the settings tab on the global, ensure the acquisition duration is set to 360 seconds.In the experiment tab, select the new list of subjects and create a new experiment file and a new list of tested subjects by following the instructions in the opened tab. Prepare the animals for the test by wrapping single-sided adhesive tape around three-fourths of the animal's tail starting from the base. Start the run by clicking start run then continue in the acquisition tab.Immediately after hanging the animal on the hook, click the start icon under the visualized position to acquire data. After testing all the animals, in the analysis tab, select the last four minutes of the acquisition for analysis. Select all valid runs in the analysis period and click analyze selected subjects.Choose the desired data format and click export selected data to export the collected data. In the tail suspension test, Ube3a mutant animals were immobile for significantly longer than their control littermates, indicating their depression-like behavior.