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

Anslagsfinansiering tillhandahölls av NIH HEAL UG3 NS123958. Bostäderna inspekterades och ackrediterades av AAALAC. Djuren inhystes i Animal Resources Center (ARC) som underhålls av laboratoriepersonalen och personalen vid avdelningen för laboratorie- och djurresurser (DLAR). Procedurerna för beteendetestning är standardmetoder inom området som godkänts av American Pain Society och International Association for the Study of Pain. Metoden för avlivning överensstämmer med rekommendationerna från panelen för eutanasi vid American Veterinary Medical Association.

Alla djurförsök som beskrivs är i enlighet med NIH:s guide för vård och användning av försöksdjur. Studierna godkändes av den lokala kommittén för institutionell vård och användning (IACUC #23-201364-HSC) vid University of New Mexico Health Sciences Center. Alla studier följer riktlinjer under överinseende av en OLAW Assurance of Compliance (A3002-01) om användning av djur i forskning, enligt beskrivningen i del III. II. Försäkringar och certifieringar. Djuren är inhysta i Animal Resources Center (ARC)...

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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>

Denna modell av kronisk ryggsmärta är enkel att framkalla, och överkänslighet som etableras inom 1 vecka kan pågå i upp till (och möjligen längre) 8 veckor. Detta möjliggör noggranna studier av det kroniska smärttillståndet i motsats till andra akuta modeller som bara varar i en vecka eller två. Medan vi visar modellen på möss, kan den uPA-inducerade CBP-modellen också etableras på råttor2. En fördel med modellen är att det förlängda tidsförloppet framkallar utvecklingen av ...

Ländryggssmärta är en av de vanligaste orsakerna till funktionsnedsättning och 1 av 5 personeri världen lider 1. Trots dessa ansträngningar är det få tillförlitliga djurmodeller av ryggsmärta som är populära i djurförsök inom smärtområdet, särskilt på möss. Tidigare modeller har nästan uteslutande använt råttor för induktion av kronisk ryggsmärta (CBP) såsom de som induceras genom injektion av urinplasminogenaktivator (uPA) i ländryggens facettled 2,3, injektion av nervtillväxtfaktor (NGF) i bålmuskulatur4 eller mononatriumjodacetat (MIA)5 eller interleukin-1beta6 injektion i den intravertebrala disken. Naturligtvis är råttor att föredra för dessa modeller, främst på grund av deras större storlek och lättillgänglighet för injektion av inflammatoriska medel.
För att vara tydlig, musmodeller av ryggsmärta finns som SPARC-null-musmodellen av intervertebral diskdegeneration som använts i många år7, men dessa är mer kostsamma och tidskrävande att etablera än injektionsbaserade modeller. En nyligen genomförd musstudie etablerade en modell för smärta i nedre delen av ryggen genom att kombinera NGF-injektion i ländryggsmuskler med vertikal kronisk fasthållningsstress8. I följande protokoll har vi anpassat den uPA-inducerade CBP-modellen från råttor för möss2. Överkänslighet etableras inom 1 vecka och kvarstår upp till 6-8 veckor. Dessutom konstaterar vi att möss utvecklar ångest- och depressionsliknande beteenden. Med tanke på förekomsten av ryggsmärta och den vanligare användningen av möss i molekylär smärtforskning, är denna hållbara modell lätt etablerad för användning i utvecklingen av nya behandlingsstrategier för lindring av ryggsmärtor.

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			<td>BALB/c mice</td>
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			<td>Anesthesia induction chamber&#160;</td>
			<td>Summit Medical Equipment Company</td>
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			<td>Autoclave</td>
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			<td>Glass bead sterilizer Germinator 500</td>
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			<td>IITC Life Sciences Series 8 Model PE34 Hot/Cold Plate Analgesia Meter</td>
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			<td>PE34</td>
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			<td>Integra Miltex cotton &#38; dressing pliers&#160;</td>
			<td>Safco Dental Supply</td>
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			<td>New York Microscope Company</td>
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			<td>Plantar Test System with InfraRed Emitter, i. e. Hargreaves Apparatus</td>
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			<td>Scalpel Handle No. 3</td>
			<td>VWR</td>
			<td>25607-947</td>
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			<td>Small animal heating pad&#160;</td>
			<td>Valley Vet Supply</td>
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			<td>Student Vannas spring scissors, straight blade&#160;</td>
			<td>Fine Science Tools</td>
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			<td>Table top animal research portable anesthesia workstation &#8220;PAM&#8221;</td>
			<td>Patterson Scientific</td>
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			<td>Von Frey Filaments</td>
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urokinase-type plasminogen activator-induced mouse back pain model

En modell för ihållande smärta i nedre delen av ryggen kan induceras hos möss med den enkla metodik som beskrivs här. Steg-för-steg-metoder för enkel, snabb induktion av en ihållande ryggsmärtmodell hos möss tillhandahålls här med hjälp av en injektion av plasminogenaktivator av urokinastyp (urokinas), ett serinproteas som finns hos människor och andra djur. Metoden för induktion av ihållande smärta i nedre delen av ryggen hos möss innebär en enkel injektion av urokinas längs ligamentområdet i ländryggen. Det urokinasinflammatoriska ämnet aktiverar plasminogen till plasmin. Vanligtvis kan modellen induceras inom 10 minuter och överkänslighet kvarstår i minst 8 veckor.
Överkänslighet, gångstörning och andra vanliga ångest- och depressionsliknande mått kan testas i den kvarstående modellen. Ryggsmärta är den vanligaste typen av smärta. För att öka medvetenheten om ryggsmärta har International Association for the Study of Pain (IASP) utsett 2021 till "Global Year about Back Pain" och 2022 till "Global Year for Translating Pain Knowledge to Practice". En begränsning för den terapeutiska utvecklingen av smärtterapi är bristen på lämpliga modeller för att testa ihållande och kronisk smärta. Funktionerna i denna modell är lämpliga för att testa potentiella terapier som syftar till att minska ryggsmärta och dess tillhörande egenskaper, vilket bidrar till IASP:s utnämning 2022 till det globala året för att översätta smärtkunskap till praktik.

Nociceptiv beteendetestning och dataanalys Åberopade åtgärder Överkänslighet på trampdynan utvecklas inom ett dygn efter urokinasinjektionen. Inom 1 vecka sänks uttagströskeln avsevärt och kvarstår fram till avlivning; Detta visas till och med vecka 4 efter operationen (Figur 4A). Tassuttagsfördröjning analyseras med hjälp av von Frey upp-ner-metoden9 och Hargreaves-testet. I det plottade exemplet visade möss med C...

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 för enkel, snabb induktion av en ryggsmärtmodell hos möss tillhandahålls här med hjälp av en intraligamentinjektion av urinplasminogenaktivator.

Plasminogenaktivatorinducerad musryggsmärta av urokinastyp

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 ...

Detta protokoll beskriver en icke-invasiv metod för att inducera kronisk ryggsmärta hos möss i upp till 10 veckor, som kan utföras snabbt och tillförlitligt med minimal träning. Den svåraste aspekten av denna teknik är att känna igen hur det känns att injicera ligament jämfört med att injicera muskler. Att känna motstånd vid kontakt indikerar ligamentinjektion.Inget motstånd innebär att nålen kan vara i muskeln eller bukhålan. Börja med att ta en steril plan yta för att placera musen. Placera sedan en återhämtningsstation för värmedynor bredvid operationsuppställningen för att omedelbart överföra möss efter injektionen.Bered sedan urokinas och en Hamilton-spruta. Rita lösningen i förväg för att minimera tiden som musen tillbringar under narkos. Efter att ha bedövat musen, använd två fingrar för att lokalisera ländryggssegmenten vid L2 och L3 under bröstkorgen och kontrollera injektionsstället.Placera spetsen på Hamilton-sprutan intill ryggraden. Placera sprutan i ungefär 45 graders vinkel i ligamentet omedelbart intill benet. För in nålspetsen försiktigt, men bestämt, i ligamentet och injicera långsamt tills alla fem mikroliter har injicerats.Ta bort nålen försiktigt och långsamt. Placera sedan musen i värmeåtervinningsstationen tills den vaknar och blir rörlig. Kontrollera mössen i en timme efter operationen för att säkerställa att all normal motorisk funktion fortsätter.Efter operationen, kontrollera mössens vikt och injektionsställe dagligen i en vecka för att förhindra infektion eller komplikationer. Före testning, flytta varje djur till ett separat genomskinligt bås på testbordet med en skärmtopp. Stimulera sedan trampdynan med 3,61 Von Frey-filamentet som framkallar fyra gram kraft.Ett tillbakadragande av tre till fem stimuli anses vara ett positivt svar. Applicera nu nästa svagare filament i serien tills djuret slutar svara på den mekaniska stimuleringen. Använd det högre filamentet tills ett svar framkallas, byt sedan tillbaka till det lägre filamentet.Med hjälp av en kurvanpassningsalgoritm beräknar du den mekaniska uttagströskeln, vilket är den minimala mängd kraft som behövs för att framkalla ett svar 50 % av tiden. För att utföra Hargreaves-testet, placera mössen i bås på en glasyta som värms upp underifrån av en infraröd sändare. Mät tiden från det att infraröd ljusstimulus appliceras på musens baktass till tillbakadragande som fotlatens.För kallplattetestet, placera mössen på kylplatteapparaten som kyls till minus nio grader Celsius. Anteckna den tid det tar för musen att lyfta foten efter att ha placerats på apparaten som latensen för att dra ut foten. För att utföra ångesttestet, placera varje djur i testboxen för platspreferens med en passage mellan två kammare.En apparatkammare ska vara starkt upplyst, medan den motsatta sidan ska vara mörk. Använd en dator för att övervaka djurets ljusa och mörka vistelsetider och övergångar under ett 10-minuterstest. Placera sedan modellgnagarna eller naiva djuren på nolllabyrinten, eller använd tvåkammartestet för ljus eller mörk platspreferens.Bedöm tiden som spenderas i stängda och öppna områden i labyrinten med hjälp av ett stoppur. För Sucrose Splash Depression Test, spraya en 10 till 30 % sackaroslösning på ryggkoden och poängsätt frekvensen, varaktigheten och latensen för grooming i 10 minuter. För att utföra nya objekttester, vänj mössen individuellt vid en genomskinlig plastbur med öppen topp i en timme.Lägg till två identiska minifigurer i motsatta hörn av buren i fem minuter. Acklimatisera djuren på testdagen i en timme och placera de två identiska minifigurerna i samma positioner i buren i fem minuter innan du sätter tillbaka dem i hemburen. Ersätt sedan en av de ursprungliga figurerna med ett helt annat nytt föremål.Efter fyra timmar sätter du tillbaka mössen i testburen och registrerar den tid som ägnas åt att utforska föremålen. För bedömning av motorisk funktion, konstruera en tunnel från ett pappershandduksrör som skärs på längden på ena kanten. Sprid ut tunneln på ett rent skrivarpapper.Håll mössen försiktigt tills de är lugna. Applicera giftfritt indiskt bläck på en muss tassar genom att placera dem i en bläckbrunn, stämpeldyna eller använda en bomullspinne. Släpp ut mössen vid tunnelmynningen.Låt dem springa genom tunneln och fånga dem i slutet. Gör tassavtryck baserat på steglängd, stegbredd och tåspridning. Mekaniska och termiska överkänslighetsanalyser visade att mössen med kronisk ryggsmärta hade signifikant ökad mekanisk, värme- och kylkänslighet jämfört med de naiva kontrollerna.Ångestanalys visade att den totala tiden som tillbringades i ljuskammaren i den ljusa och mörka lådan och antalet uppfödningstillfällen var signifikant lägre hos möss med kronisk ryggsmärta, vilket tyder på ångest. Under Sucrose Splash Test uppvisade möss med kronisk ryggsmärta lägre putsningstid och antalet putsningar. Tiden innan pälsvården påbörjas ökade dock avsevärt.Steglängden mellan möss med kronisk ryggsmärta och naiva möss skilde sig markant. Det viktigaste att komma ihåg när du försöker denna procedur är att du siktar på ligamenten runt ryggraden, inte muskeln. Modellen för kronisk ryggsmärta ger ett tillförlitligt verktyg för att bedöma en mängd ny forskning, farmakologisk och andra potentiella terapier.

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

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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.

Bedöma Functional Performance i<em&gt; Mdx</em&gt; Musmodell

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.

Fastställande Smärta Detection och toleranströsklar hjälp av en integrerad, multimodal 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 av ländryggsbesvär misslyckad ryggkirurgi hos Multicolumn-bly ryggmärgsstimulering

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.

Mätning av aktiv och passiv tamhet separat hos möss

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-ischemia smärt modell för komplext regionalt smärtsyndrom typ-i hos råttor

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).

Undersöka smärtrelaterat undvikande beteende med hjälp av ett robotarms-nå 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 ...

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.

Ryggmekanisk känslighetsbedömning hos råtta för mekanistisk undersökning av kronisk ryggsmärta

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.

Förhöjt plus labyrinttest i en musmodell med Angelmans syndrom

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.

Öppet fälttest i en musmodell med Angelmans syndrom

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.