Name: back mechanical sensitivity assessment in the rat for mechanistic investigation of chronic back pain
Uploaded: 2022-08-30T13:00:00
Description: Watch this Scientific Journal Video about Back Mechanical Sensitivity Assessment in the Rat for Mechanistic Investigation of Chronic Back Pain at JoVE.com

Detta arbete stöddes av ett bidrag från Fondation Chiropratique du Québec och Natural Sciences and Engineering Research Council of Canada (MP: grant #06659). HK: s bidrag stöddes av Université du Québec à Trois-Rivières (PAIR-programmet). BP:s bidrag stöddes av Fonds de recherche du Québec en Santé (FRQS) och Fondation Chiropratique du Québec. TP: s bidrag stöddes av Natural Sciences and Engineering Research Council of Canada. NE:s och EK:s bidrag stöddes av Fondation Chiropratique du Québec. MP:s bidrag stöddes av FRQS.

Försöksprotokollet godkändes av djurvårdskommittén vid Université du Québec à Trois-Rivières och överensstämde med riktlinjerna från Canadian Council on Animal Care och riktlinjerna från kommittén för forskning och etiska frågor vid International Association for the Study of Pain (IASP). I den aktuella studien användes sex Wistar-hanråttor (kroppsvikt: 320-450 g; ålder: 18-22 veckor). Djuren erhölls från en kommersiell källa (se Materialförteckning). Data från dessa råttor är fr...

<ol>
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Kritiska steg BMS-testet är en enkel metod för att bedöma mekanisk känslighet i ryggen på råttor, antingen vid en tidpunkt eller upprepade gånger under dagar eller veckor, när förändringar förväntas inträffa (smärtmodeller) eller efter farmakologisk eller icke-farmakologisk intervention. Kritiska frågor om metoden inkluderar testburen, vars dimensioner måste säkerställa att råttan är bekväm men inte rör sig för mycket. Djurets rygg måste vara tillgänglig genom nättaket för r...

Ländryggssmärta (LBP) är den främsta orsaken till funktionshinder över hela världen, vilket har dramatiska personliga, ekonomiska och sociala konsekvenser 1,2,3,4. Varje år drabbas cirka 37% av befolkningen av LBP5. LBP försvinner vanligtvis inom några veckor men återkommer hos 24% -33% av individerna och blir kronisk i 5% -10% av fallen2. För att förstå mekanismerna och effekterna av LBP samt effekterna av olika terapeutiska ingrepp har flera djurmodeller av LBP använts, som efterliknar kliniska tillstånd eller vissa komponenter av LBP6. Dessa mus- och råttmodeller kan klassificeras i en eller flera av följande kategorier: (1) diskogen LBP7,8,, (2) radikulär LBP 8,9,10,11, (3) fasettledsartros 12 och (4) muskelinducerad LBP 13,14 . Eftersom smärtan inte kan mätas direkt i icke-mänskliga arter har många tester utvecklats för att kvantifiera smärtliknande beteenden i dessa modeller8. Dessa tester bedömer beteenden som framkallas av en skadlig stimulans (mekanisk kraft 15,16,17, termisk stimulering 18,19,20,21,22,23,24,25) eller produceras spontant26,27,28,29.
Metoderna som använder mekaniska stimuli inkluderar Von Frey-testet 15,16 och Randall-Selitto-testet17. Metoder som använder värmestimuli inkluderar svansflicktestet18, värmeplatttestet19, Hargreaves-testet20 och termiskt sondtest21. Metoder som använder kalla stimuli inkluderar kallplatttestet 22, acetonavdunstningstestet23 och kallplantaranalysen24. Metoder för spontana beteenden inkluderar grimasskalorna 26, grävning27, viktbärande och gånganalys 28, samt en automatiserad beteendeanalys29. Trots dessa många tillgängliga tester är ingen av dem utformade speciellt för ryggsmärtmodeller.
Syftet med denna studie är att fastställa ett enkelt och tillgängligt test för att bedöma mekanisk känslighet i ryggen på råttor. Tekniken bygger till stor del på Von Frey-testet som tillämpas på baktassens plantaryta15,16. Grundprincipen för Von Frey-testet är att använda en serie monofilament till intresseområdet och leverera konstanta förutbestämda krafter. Ett svar anses vara positivt om råttan visar ett nocifensivt beteende. Den mekaniska tröskeln kan sedan beräknas baserat på filamenten som framkallade svar. I den aktuella studien ges en enkel och tillgänglig metod anpassad från Von Frey-testet för att bestämma mekanisk känslighet i ryggen på råttor.

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	<tbody>
		<tr>
			<td>Aerrane (isoflurane, USP) - Veterinary Use Only</td>
			<td>Baxter</td>
			<td>NDC 10019-773-60</td>
			<td>Inhalation Anaesthetic ; DIN 02225875, for inducing anasthesia</td>
		</tr>
		<tr>
			<td>Complete Freund Adjuvant (CFA)</td>
			<td>Fisher Scientific</td>
			<td>#77140</td>
			<td>Water-in-oil emulsion of Complete Freund Adjuvant (CFA) with killed cells of Mycobacterium butyricum.</td>
		</tr>
		<tr>
			<td>Male Wistar Rats</td>
			<td>Charles River Laboratories</td>
			<td></td>
			<td>body weight: 320&#8211;450 g; age: 18-22 weeks.</td>
		</tr>
		<tr>
			<td>Penlon Sigma Delta Vaporizer</td>
			<td>Penlon</td>
			<td>990-VI5K-SVEEK</td>
			<td>Penlon Sigma Delta Vaporizer used for anasthesia</td>
		</tr>
		<tr>
			<td>Sharpie Permanent Marker</td>
			<td>Sharpie</td>
			<td>BC23636</td>
			<td>Permanent Marker, Fine Point, Black</td>
		</tr>
		<tr>
			<td>Test cage</td>
			<td>Custom-made</td>
			<td></td>
			<td>Width: 20 cm;&#160; Length: 50 cm; Height from the bottom to the top: 40 cm; Height from the bottom mesh to the top of the cage: 7 cm; Wall thickness: 5 mm; Mesh: 1 mm wire with an 8 mm inter-wire distance&#160;&#160;&#160;</td>
		</tr>
		<tr>
			<td>Von Frey Filaments</td>
			<td>Aesthesio, Precise Tactile Sensory Evaluator</td>
			<td>514000-20C</td>
			<td>Filaments from 0.07 g to 26 g</td>
		</tr>
		<tr>
			<td>Wahl Professional Animal, ARCO Cordless Pet Clipper, Trimmer Grooming&#160;</td>
			<td>Wahl</td>
			<td>Kit #8786-1201</td>
			<td>Animal hair trimmer, for shaving purposes, zero blade&#160;</td>
		</tr>
	</tbody>
</table>

back mechanical sensitivity assessment in the rat for mechanistic investigation of chronic back pain

Ländryggssmärta är den främsta orsaken till funktionshinder över hela världen, med dramatiska personliga, ekonomiska och sociala konsekvenser. För att utveckla nya terapier behövs djurmodeller för att undersöka mekanismerna och effektiviteten hos nya terapier ur ett translationellt perspektiv. Flera gnagarmodeller av ryggsmärta används i aktuella undersökningar. Överraskande validerades dock inget standardiserat beteendetest för att bedöma mekanisk känslighet i ryggsmärtmodeller. Detta är avgörande för att bekräfta att djur med förmodad ryggsmärta uppvisar lokal överkänslighet mot nociceptiva stimuli och för att övervaka känslighet under interventioner som är utformade för att lindra ryggsmärta. Syftet med denna studie är att fastställa ett enkelt och tillgängligt test för att bedöma mekanisk känslighet i ryggen på råttor. En testbur tillverkades speciellt för denna metod; Längd x bredd x höjd: 50 x 20 x 7 cm, med ett rostfritt stålnät på ovansidan. Denna testbur möjliggör applicering av mekaniska stimuli på baksidan. För att utföra testet rakas djurets baksida i det intressanta området, och testområdet markeras för att upprepa testet på olika dagar, efter behov. Den mekaniska tröskeln bestäms med Von Frey-filament applicerade på paraspinalmusklerna, med användning av upp-ner-metoden som beskrivits tidigare. De positiva svaren inkluderar (1) muskelryckningar, (2) välvning (ryggförlängning), (3) rotation av nacken (4) skrapa eller slicka ryggen och (5) fly. Detta beteendetest (Back Mechanical Sensitivity (BMS) -test) är användbart för mekanistisk forskning med gnagarmodeller av ryggsmärta för utveckling av terapeutiska ingrepp för förebyggande och hantering av ryggsmärta.

Metoden användes i en tidigare studie, där fullständiga data och statistik presenterades för att jämföra tillbaka mekanisk känslighet mellan CFA och kontrollråttor30. Representativa individuella data (medelvärde av vänster och höger tröskelvärden) från sex råttor som ingick i föregående studie presenteras i figur 3 och tabell 1. Vid baseline var den mekaniska känsligheten likartad mellan grupperna. Intramuskulär injektion av CFA i l...

Watch this Scientific Journal Video about Back Mechanical Sensitivity Assessment in the Rat for Mechanistic Investigation of Chronic Back Pain at JoVE.com

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

För att utveckla nya terapeutiska interventioner för förebyggande och hantering av ryggsmärta krävs djurmodeller för att undersöka mekanismerna och effektiviteten hos dessa terapier ur ett translationellt perspektiv. Detta protokoll beskriver BMS-testet, en standardiserad metod för att bedöma mekanisk känslighet hos råtta.

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, ...

Detta protokoll är användbart för att testa mekanisk känslighet i ryggen. Det är särskilt intressant för longitudinella mönster i råttmodeller av kronisk ryggsmärta, för vilka målet är att undersöka effektiviteten av en intervention för att minska ryggkänsligheten för skadliga stimuli. Metoden är enkel, tillgänglig och ger reproducerbara resultat i utbildade experimenter.Det är till stor del baserat på Von Frey-testet, som vanligtvis används för att bedöma mekanisk känslighet vid bakpoten. Som för alla andra beteendetest bör experimenterna öva på att standardisera tillämpningen av filamenten, och tills beteenden som produceras av mekaniska stimuli lätt skiljer sig från andra beteenden. Testet är enkelt, annars, och det borde inte vara några större svårigheter att använda det.Demonstrerar proceduren kommer att vara Nasim Eskandari, doktorand från mitt laboratorium. För att inducera kronisk ryggsmärta hos råtta, utför en intramuskulär injektion av Complete Freunds Adjuvans, eller CFA, i ryggmusklerna. Använd en 27 gauge nål, injicera 150 mikroliter av en färdig att använda vatten-i-olja emulsion av CFA i paraspinalmusklerna ensidigt eller bilateralt.Håll nålen på plats i minst tre minuter. Därefter tillverkar du testburet för två djur, bestående av en kammare för varje djur genom att montera de två angränsande kamrarna på fyra 33 centimeter långa plexiglasben. Använd transparent plexiglas för kamrarnas väggar och svart plexiglas för att separera kamrar för att förhindra att djur ser varandra.Gör sedan golvet och taket i testburet med rostfritt stålnät av en millimeter tråd med ett åtta millimeter interwire-avstånd. Bekanta råttan med testburen i 30 minuter per dag under fem till sju dagar i följd före det första testet. I en benägen position under isofluranbedövning, raka rygghåret i intresseområdet över cirka 10 kotnivåer med hjälp av en djurhårtrimmer.För upprepade åtgärder, raka rygghåret var tredje dag utan beteendebedömning för att säkerställa att stimuli alltid appliceras direkt på huden. Rita ett märke på huden med en permanent markör för att säkerställa att filament alltid appliceras på samma område när testet upprepas på olika dagar. På testdagen placeras råttan i testburen i 15–30 minuter före testet tills råttan är lugn.Applicera sedan Von Frey-filament vinkelrätt mot baksidan, börja alltid med tvågramfilamentet och använd upp/ner-metoden. Applicera filamentet i två sekunder bilateralt till intresseområdet 10 millimeter från ryggraden bearbetas var 15 till 30 sekunder när råttan är vaken, står på sina fyra tassar och inte rör sig. Om inget svar observeras vid applicering av ett filament, applicera nästa filament med en högre kraft i serien.Om ett positivt svar som att böja ryggen, slicka, muskelryckningar, flytta bort från filamentet eller rotation observeras, använd nästa filament med en lägre kraft i serien. Fortsätt proceduren tills fyra avläsningar erhålls efter den första beteendeförändringen. I denna studie undersöktes ryggmekanisk känslighet mellan CFA och kontrollråttor.Vid baseline var den mekaniska känsligheten likartad mellan grupperna. Intramuskulär injektion av CFA i ländryggsmusklerna orsakade en markant ökning av mekanisk känslighet från sju till 28 dagar. Däremot visade kontrollråttorna ingen signifikant förändring.Vidare bekräftade histologisk undersökning förekomsten av kroniska inflammatoriska förändringar i musklerna injicerade med CFA med en tydlig leukocytinfiltration. Friska ryggmuskler från en kontrollerad råtta efter intramuskulär injektion av saltlösning visas som referens. Den mekaniska överkänsligheten observerades vid bakpoten med ett standard Von Frey-test, förutom ryggmusklerna.Det är mycket viktigt att se till att filamentet alltid appliceras på samma sätt vinkelrätt mot ryggen och att djuret inte rör sig när det appliceras. Experimenten bör också känna förtroende för att skilja beteendemässiga svar för moderna rörelser.

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Research

JoVE Journal

Behavior

Use of the Operant Orofacial Pain Assessment Device (OPAD) to Measure Changes in Nociceptive Behavior

We present an operant system for the detection of pain in awake, conscious rodents. The Orofacial Pain Assessment Device (OPAD) assesses pain behaviors in a more clinically relevant way by not relying on reflex-based measures of nociception. Food fasted, hairless (or shaved) rodents are placed into a Plexiglas chamber which has two Peltier-based thermodes that can be programmed to any temperature between 7 &deg;C and 60 &deg;C. The rodent is trained to make contact with these in order to access a reward bottle. During a session, a number of behavioral pain outcomes are automatically recorded and saved. These measures include the number of reward bottle activations (licks) and facial contact stimuli (face contacts), but custom measures like the lick/face ratio (total number of licks per session/total number of contacts) can also be created. The stimulus temperature can be set to a single temperature or multiple temperatures within a session. The OPAD is a high-throughput, easy to use operant assay which will lead to better translation of pain research in the future as it includes cortical input instead of relying on spinal reflex-based nociceptive assays.

Use of the Operant Orofacial Pain Assessment Device OPAD to Measure Changes in Nociceptive Behavior

We present a user-friendly, high-throughput operant system for the evaluation of pain behaviors in awake, conscious rodents. The Orofacial Pain Assessment Device (OPAD) can assess pain through a reward/conflict paradigm thus providing a more humane way of testing. This protocol will yield more clinically relevant and translational data from rodents.

Användning av Operant Orofacial Pain Assessment Device (OPAD) för att mäta förändringar i Nociceptive Behavior

We present an operant  system for the detection of pain in awake, conscious rodents. The Orofacial  Pain Assessment Device (OPAD) assesses pain behaviors ...

A Protocol of Manual Tests to Measure Sensation and Pain in Humans

Numerous qualitative and quantitative techniques can be used to test sensory nerves and pain in both research and clinical settings. The current study demonstrates a quantitative sensory testing protocol using techniques to measure tactile sensation and pain threshold for pressure and heat using portable and easily accessed equipment. These techniques and equipment are ideal for new laboratories and clinics where cost is a concern or a limiting factor. We demonstrate measurement techniques for the following: cutaneous mechanical sensitivity on the arms and legs (von-Frey filaments), radiant and contact heat sensitivity (with both threshold and qualitative assessments using the Visual Analog Scale (VAS)), and mechanical pressure sensitivity (algometer, with both threshold and the VAS). The techniques and equipment described and demonstrated here can be easily purchased, stored, and transported by most clinics and research laboratories around the world. A limitation of this approach is a lack of automation or computer control. Thus, these processes can be more labor intensive in terms of personnel training and data recording than the more sophisticated equipment. We provide a set of reliability data for the demonstrated techniques. From our description, a new laboratory should be able to set up and run these tests and to develop their own internal reliability data.

The goal of this procedure is to demonstrate a battery of quantitative techniques for sensory and pain measurement in humans. The equipment and techniques described are commonly found in pain clinics or are easy to obtain.

Ett protokoll manuella test för att mäta Sensation och smärtsamma för människor

Numerous qualitative and quantitative techniques can be used to test sensory nerves and pain in both research and clinical settings. The current study ...

Novel Object Recognition Test for the Investigation of Learning and Memory in Mice

The object recognition test (ORT) is a commonly used behavioral assay for the investigation of various aspects of learning and memory in mice. The ORT is fairly simple and can be completed over 3 days: habituation day, training day, and testing day. During training, the mouse is allowed to explore 2 identical objects. On test day, one of the training objects is replaced with a novel object. Because mice have an innate preference for novelty, if the mouse recognizes the familiar object, it will spend most of its time at the novel object. Due to this innate preference, there is no need for positive or negative reinforcement or long training schedules. Additionally, the ORT can also be modified for numerous applications. The retention interval can be shortened to examine short-term memory, or lengthened to probe long-term memory. Pharmacological intervention can be used at various times prior to training, after training, or prior to recall to investigate different phases of learning (i.e., acquisition, early or late consolidation, or recall). Overall, the ORT is a relatively low-stress, efficient test for memory in mice, and is appropriate for the detection of neuropsychological changes following pharmacological, biological, or genetic manipulations.

The object recognition test (ORT) is a simple and efficient assay for evaluating learning and memory in mice. The methodology is described below.

Nya objekt erkännande Test för utredning av inlärning och minne hos möss

The object recognition test (ORT) is a commonly used behavioral assay for the investigation of various aspects of learning and memory in mice. The ORT is ...

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 ...

The Unpredictable Chronic Mild Stress Protocol for Inducing Anhedonia in Mice

Depression is a highly prevalent and debilitating condition, only partially addressed by current pharmacotherapies. The lack of response to treatment by many patients prompts the need to develop new therapeutic alternatives and to better understand the etiology of the disorder. Pre-clinical models with translational merits are rudimentary for this task. Here we present a protocol for the unpredictable chronic mild stress (UCMS) method in mice. In this protocol, adolescent mice are chronically exposed to interchanging unpredictable mild stressors. Resembling the pathogenesis of depression in humans, stress exposure during the sensitive period of mice adolescence instigates a depressive-like phenotype evident in adulthood. UCMS can be used for screenings of antidepressants on the variety of depressive-like behaviors and neuromolecular indices. Among the more prominent tests to assess depressive-like behavior in rodents is the sucrose preference test (SPT), which reflects anhedonia (core symptom of depression). The SPT will also be presented in this protocol. The ability of UCMS to induce anhedonia, instigate long-term behavioral deficits and enable reversal of these deficits via chronic (but not acute) treatment with antidepressants strengthens the protocol&#39;s validity compared to other animal protocols for inducing depressive-like behaviors.

Here we present the unpredictable chronic mild stress protocol in mice. This protocol induces a long-term depressive-like phenotype and enables to assess the efficacy of putative antidepressants in reversing the behavioral and neuromolecular depressive-like deficits.

Oförutsägbara kronisk Mild Stress protokollet för att inducera likgiltighet hos möss

Depression is a highly prevalent and debilitating condition, only partially addressed by current pharmacotherapies. The lack of response to treatment by ...

Multi-Modal Signals for Analyzing Pain Responses to Thermal and Electrical Stimuli

The assessment of pain relies mostly on methods that require a person to communicate. However, for people with cognitive and verbal impairments, existing methods are not sufficient as they lack reliability and validity. To approach this problem, recent research focuses on an objective pain assessment facilitated by parameters of responses derived from physiology, and video and audio signals. To develop reliable automated pain recognition systems, efforts have been made in creating multimodal databases in order to analyze pain and detect valid pain patterns. While the results are promising, they only focus on discriminating pain or pain intensities versus no pain. In order to advance this, research should also consider the quality and duration of pain as they provide additional valuable information for more advanced pain management. To complement existing databases and the analysis of pain regarding quality and length, this paper proposes a psychophysiological experiment to elicit, measure, and collect valid pain reactions. Participants are subjected to painful stimuli that differ in intensity (low, medium, and high), duration (5 s / 1 min), and modality (heat / electric pain) while audio, video (e.g., facial expressions, body gestures, facial skin temperature), and physiological signals (e.g., electrocardiogram [ECG], skin conductance level [SCL], facial electromyography [EMG], and EMG of M. trapezius) are being recorded. The study consists of a calibration phase to determine a subject&#8217;s individual pain range (from low to intolerable pain) and a stimulation phase in which pain stimuli, depending on the calibrated range, are applied. The obtained data may allow refining, improving, and evaluating automated recognition systems in terms of an objective pain assessment. For further development of such systems and to investigate pain reactions in more detail, additional pain modalities such as pressure, chemical, or cold pain should be included in future studies. Recorded data of this study will be released as the &#8220;X-ITE Pain Database&#8221;.

This article focuses on the experimental elicitation of pain through heat (thermal) and electrical stimulation while recording physiological, visual, and paralinguistic responses. It aims at collecting valid multimodal data for analyzing pain based on its intensity, quality, and duration.&#160;

Multimodala signaler för att analysera smärta Svaren till termisk och elektrisk Stimuli

The assessment of pain relies mostly on methods that require a person to communicate. However, for people with cognitive and verbal impairments, existing ...

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 ...

Assessment of Memory Function in Pilocarpine-induced Epileptic Mice

Cognitive impairment is one of the most common comorbidities in temporal lobe epilepsy. To recapitulate epilepsy-associated cognitive decline in an animal model of epilepsy, we generated pilocarpine-treated chronic epileptic mice. We present a protocol for three different behavioral tests using these epileptic mice: novel object location (NL), novel object recognition (NO), and pattern separation (PS) tests to evaluate learning and memory for places, objects, and contexts, respectively. We explain how to set the behavioral apparatus and provide experimental procedures for the NL, NO, and PS tests following an open field test that measures the animals&#8217; basal locomotor activities. We also describe the technical advantages of the NL, NO, and PS tests with respect to other behavioral tests for assessing memory function in epileptic mice. Finally, we discuss possible causes and solutions for epileptic mice failing to make 30 s of good contact with the objects during the familiarization sessions, which is a critical step for successful memory tests. Thus, this protocol provides detailed information about how to assess epilepsy-associated memory impairments using mice. The NL, NO, and PS tests are simple, efficient assays that are appropriate for the evaluation of different kinds of memory in epileptic mice.

This article presents experimental procedures for assessing memory impairments in pilocarpine-induced epileptic mice. This protocol can be used to study the pathophysiologic mechanisms of epilepsy-associated cognitive decline, which is one of the most common comorbidities in epilepsy.

Bedömning av minnesfunktion i Pilocarpin-inducerade epileptiska möss

Cognitive impairment is one of the most common comorbidities in temporal lobe epilepsy. To recapitulate epilepsy-associated cognitive decline in an animal ...

Chronic Stress Shifts Effort-Related Choice Behavior in a Y-Maze Barrier Task in Mice

Mood disorders, including major depressive disorder, can be precipitated by chronic stress. The Y-maze barrier task is an effort-related choice test that measures motivation to expend effort and obtain reward. In mice, chronic stress exposure significantly impacts motivation to work for a higher value reward when a lesser value reward is freely available compared to unstressed mice. Here we describe the chronic corticosterone administration paradigm, which produces a shift in effortful responding in the Y-maze barrier task. In the Y-maze task, one arm contains 4 food pellets, while the other arm contains only 2 pellets. After mice learn to select the high reward arm, barriers with progressively increasing height are then introduced into the high reward arm over multiple test sessions. Unfortunately, most chronic stress paradigms (including corticosterone and social defeat) were developed in male mice and are less effective in female mice. Therefore, we also discuss chronic non-discriminatory social defeat stress (CNSDS), a stress paradigm we developed that is effective in both male and female mice. Repeating results with multiple distinct chronic stressors in male and female mice combined with increased usage of translationally relevant behavior tasks will help to advance the understanding of how chronic stress can precipitate mood disorders.

The Y-maze barrier task is a behavior test that examines motivation to expend effort for reward. Here, we discuss testing multiple well-validated chronic stressors including chronic corticosterone and social defeat stress with this behavior, as well as the novel chronic non-discriminatory social defeat stress (CNSDS), which is effective in females.

Kronisk stress Skift Ansträngning-Relaterade Choice Beteende i en Y-Maze Barrier Uppgift hos möss

Mood disorders, including major depressive disorder, can be precipitated by chronic stress. The Y-maze barrier task is an effort-related choice test that ...

An Improved Assay and Tools for Measuring Mechanical Nociception in Drosophila Larvae

Published assays for mechanical nociception in Drosophila have led to variable assessments of behavior. Here, we fabricated, for use with Drosophila larvae, customized metal nickel-titanium alloy (nitinol) filaments. These mechanical probes are similar to the von Frey filaments used in vertebrates to measure mechanical nociception. Here, we demonstrate how to make and calibrate these mechanical probes and how to generate a full behavioral dose-response from subthreshold (innocuous or non-noxious range) to suprathreshold (low to high noxious range) stimuli. To demonstrate the utility of the probes, we investigated tissue damage-induced hypersensitivity in Drosophila larvae. Mechanical allodynia (hypersensitivity to a normally innocuous mechanical stimulus) and hyperalgesia (exaggerated responsiveness to a noxious mechanical stimulus) have not yet been established in Drosophila larvae. Using mechanical probes that are normally innocuous or probes that typically elicit an aversive behavior, we found that Drosophila larvae develop mechanical hypersensitization (both allodynia and hyperalgesia) after tissue damage. Thus, the mechanical probes and assay that we illustrate here will likely be important tools to dissect the fundamental molecular/genetic mechanisms of mechanical hypersensitivity.

An Improved Assay and Tools for Measuring Mechanical Nociception in Drosophila Larvae

The goal of this protocol is to show how to perform an improved assay for mechanical nociception in Drosophila larvae. We use the assay here to demonstrate that mechanical hypersensitivity (allodynia and hyperalgesia) exists in Drosophila larvae.

En förbättrad analys och verktyg för att mäta mekanisk nociception i Drosophila Larver

Published assays for mechanical nociception in Drosophila have led to variable assessments of behavior. Here, we fabricated, for use with Drosophila ...