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

Dit werk werd ondersteund door een subsidie van de Fondation Chiropratique du Québec en de Natural Sciences and Engineering Research Council of Canada (MP: grant #06659). De bijdrage van HK werd ondersteund door de Université du Québec à Trois-Rivières (PAIR-programma). De bijdrage van BP werd ondersteund door het Fonds de recherche du Québec en Santé (FRQS) en de Fondation Chiropratique du Québec. De bijdrage van TP werd ondersteund door de Natural Sciences and Engineering Research Council of Canada. De bijdrage van NE en EK werd ondersteund door de Fondation Chiropratique du Québec. De bijdrage van MP werd ondersteund door de FRQS.

Het experimentele protocol werd goedgekeurd door de dierverzorgingscommissie van de Université du Québec à Trois-Rivières en voldeed aan de richtlijnen van de Canadian Council on Animal Care en de richtlijnen van de Committee for Research and Ethical Issues van de International Association for the Study of Pain (IASP). De huidige studie gebruikte zes mannelijke Wistar-ratten (lichaamsgewicht: 320-450 g; leeftijd: 18-22 weken). De dieren werden verkregen uit een commerciële bron (zie tabel met materialen</str...

<ol>
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Kritieke stappen De BMS-test is een eenvoudige methode om de mechanische gevoeligheid in de rug van ratten te beoordelen, hetzij op één tijdstip of herhaaldelijk gedurende dagen of weken, wanneer veranderingen worden verwacht (pijnmodellen) of na farmacologische of niet-farmacologische interventie. Kritieke problemen van de methode zijn de testkooi, waarvan de afmetingen ervoor moeten zorgen dat de rat comfortabel is, maar niet te veel beweegt. De rug van het dier moet toegankelijk blijven via het g...

Lage rugpijn (LBP) is wereldwijd de belangrijkste oorzaak van invaliditeit, die dramatische persoonlijke, economische en sociale gevolgen heeft 1,2,3,4. Elk jaar wordt ongeveer 37% van de bevolking getroffen door LBP5. LBP verdwijnt meestal binnen een paar weken, maar komt terug bij 24% -33% van de individuen en wordt chronisch in 5% -10% van de gevallen2. Om de mechanismen en effecten van LBP en de effecten van verschillende therapeutische interventies te begrijpen, zijn verschillende diermodellen van LBP gebruikt, die klinische aandoeningen of sommige componenten van LBP6 nabootsen. Deze muis- en rattenmodellen kunnen worden ingedeeld in een of meer van de volgende categorieën: (1) discogene LBP7,8,, (2) radiculaire LBP 8,9,10,11, (3) facetgewrichtartrose 12 en (4) spiergeïnduceerde LBP13,14 . Omdat de pijn niet direct kan worden gemeten bij niet-menselijke soorten, zijn er talloze tests ontwikkeld om pijnachtig gedrag in deze modellen te kwantificeren8. Deze tests beoordelen gedrag dat wordt opgeroepen door een schadelijke stimulus (mechanische kracht 15,16,17, thermische stimulatie 18,19,20,21,22,23,24,25) of spontaan geproduceerd 26,27,28,29.
De methoden met mechanische stimuli omvatten de Von Frey-test 15,16 en de Randall-Selitto-test17. Methoden met behulp van warmtestimuli omvatten de staartfliktest18, hete plaattest19, Hargreaves-test20 en thermische sondetest21. Methoden met koude stimuli omvatten de koude plaattest22, acetonverdampingstest 23 en koude plantaire test24. Methoden voor spontaan gedrag omvatten de grimasschalen 26, graven27, gewichtdragende en loopanalyse 28, evenals een geautomatiseerde gedragsanalyse29. Ondanks deze talrijke beschikbare tests, is geen van hen specifiek ontworpen voor rugpijnmodellen.
Het doel van deze studie is om een eenvoudige en toegankelijke test te leggen om de mechanische gevoeligheid in de rug van ratten te beoordelen. De techniek is grotendeels gebaseerd op de Von Frey-test toegepast op het plantaire oppervlak van de achterpoot15,16. Het basisprincipe van de Von Frey-test is om een reeks monofilamenten te gebruiken voor het gebied van belang, waarbij constante vooraf bepaalde krachten worden geleverd. Een reactie wordt als positief beschouwd als de rat een nocifensief gedrag vertoont. De mechanische drempel kan dan worden berekend op basis van de filamenten die reacties oproepen. In de huidige studie wordt een eenvoudige en toegankelijke methode aangeboden die is aangepast aan de Von Frey-test om de mechanische gevoeligheid in de rug van ratten te bepalen.

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

Lage rugpijn is wereldwijd de belangrijkste oorzaak van invaliditeit, met dramatische persoonlijke, economische en sociale gevolgen. Om nieuwe therapieën te ontwikkelen, zijn diermodellen nodig om de mechanismen en effectiviteit van nieuwe therapieën vanuit een translationeel perspectief te onderzoeken. Verschillende knaagdiermodellen van rugpijn worden gebruikt in lopende onderzoeken. Verrassend genoeg werd er echter geen gestandaardiseerde gedragstest gevalideerd om de mechanische gevoeligheid in rugpijnmodellen te beoordelen. Dit is van cruciaal belang om te bevestigen dat dieren met veronderstelde rugpijn lokale overgevoeligheid voor nociceptieve stimuli vertonen en om de gevoeligheid te controleren tijdens interventies die zijn ontworpen om rugpijn te verlichten. Het doel van deze studie is om een eenvoudige en toegankelijke test te leggen om de mechanische gevoeligheid in de rug van ratten te beoordelen. Speciaal voor deze methode is een testkooi vervaardigd; lengte x breedte x hoogte: 50 x 20 x 7 cm, met een roestvrijstalen gaas aan de bovenkant. Deze testkooi maakt het mogelijk om mechanische stimuli aan de achterkant toe te passen. Om de test uit te voeren, wordt de achterkant van het dier geschoren in het gebied van belang en wordt het testgebied gemarkeerd om de test op verschillende dagen te herhalen, indien nodig. De mechanische drempel wordt bepaald met Von Frey-filamenten aangebracht op de paraspinale spieren, met behulp van de eerder beschreven up-down-methode. De positieve reacties omvatten (1) spiertrekkingen, (2) boogvorming (rugverlenging), (3) rotatie van de nek (4) krabben of likken aan de rug en (5) ontsnappen. Deze gedragstest (Back Mechanical Sensitivity (BMS) test) is nuttig voor mechanistisch onderzoek met knaagdiermodellen van rugpijn voor de ontwikkeling van therapeutische interventies voor de preventie en behandeling van rugpijn.

De methode werd gebruikt in een eerdere studie, waarin volledige gegevens en statistieken werden gepresenteerd om de mechanische gevoeligheid tussen CFA en controleratten te vergelijken30. Representatieve individuele gegevens (gemiddelde van linker- en rechterdrempels) van zes ratten die in het vorige onderzoek zijn opgenomen, zijn weergegeven in figuur 3 en tabel 1. Bij baseline was de mechanische gevoeligheid vergelijkbaar tussen de groepen. Intramu...

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

Om nieuwe therapeutische interventies te ontwikkelen voor de preventie en het beheer van rugpijn, zijn diermodellen nodig om de mechanismen en effectiviteit van deze therapieën vanuit een translationeel perspectief te onderzoeken. Het huidige protocol beschrijft de BMS-test, een gestandaardiseerde methode om de mechanische gevoeligheid van de rug bij de rat te beoordelen.

Beoordeling van de mechanische gevoeligheid van de rug bij de rat voor mechanistisch onderzoek naar chronische rugpijn

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

Dit protocol is handig om de mechanische gevoeligheid aan de achterkant te testen. Het is vooral interessant voor longitudinale ontwerpen in rattenmodellen van chronische rugpijn, waarvoor het doel is om de effectiviteit van een interventie te onderzoeken om de ruggevoeligheid voor schadelijke stimuli te verminderen. De methode is eenvoudig, toegankelijk en levert reproduceerbare resultaten op bij getrainde experimentatoren.Het is grotendeels gebaseerd op de Von Frey-test, die vaak wordt gebruikt om de mechanische gevoeligheid aan de achterpoot te beoordelen. Zoals voor elke andere gedragstest, moeten de experimentatoren oefenen met het standaardiseren van de toepassing van de filamenten, en totdat gedrag geproduceerd door mechanische stimuli gemakkelijk te onderscheiden is van ander gedrag. De test is eenvoudig, anders, en er zou geen grote moeilijkheid moeten zijn om het te gebruiken.Nasim Eskandari, een promovendus uit mijn laboratorium, demonstreert de procedure. Om chronische rugpijn bij de rat te veroorzaken, voert u een intramusculaire injectie van Complete Freund's Adjuvant of CFA uit in de rugspieren. Injecteer met behulp van een naald van 27 gauge 150 microliter van een kant-en-klare water-in-olie-emulsie van CFA eenzijdig of bilateraal in de paraspinale spieren.Houd de naald minstens drie minuten op zijn plaats. Fabriceer vervolgens de testkooi voor twee dieren, bestaande uit één kamer voor elk dier door de twee aaneengesloten kamers op vier 33 centimeter lange plexiglas poten te monteren. Gebruik transparant plexiglas voor de wanden van de kamers en zwart plexiglas om kamers te scheiden om te voorkomen dat dieren elkaar zien.Maak vervolgens de vloer en het plafond van de testkooi met behulp van roestvrijstalen gaas gemaakt van één millimeter draad met een afstand van acht millimeter tussen de draad. Maak de rat vertrouwd met de testkooi gedurende 30 minuten per dag gedurende vijf tot zeven opeenvolgende dagen vóór de eerste test. Scheer in buikligging onder isofluraananesthesie het rughaar in het interessegebied over ongeveer 10 wervelniveaus met behulp van een trimmer voor dierenhaar.Voor herhaalde maatregelen, scheer het rughaar om de drie dagen zonder gedragsbeoordeling om ervoor te zorgen dat stimuli altijd rechtstreeks op de huid worden aangebracht. Teken een markering op de huid met een permanente marker om ervoor te zorgen dat filamenten altijd op hetzelfde gebied worden aangebracht wanneer de test op verschillende dagen wordt herhaald. Plaats de rat op de testdag 15 tot 30 minuten voor de test in de testkooi totdat de rat kalm is.Breng vervolgens Von Frey-filamenten loodrecht op de achterkant aan, altijd beginnend met het filament van twee gram en met behulp van de up / down-methode. Breng de gloeidraad gedurende twee seconden bilateraal aan op het interessegebied 10 millimeter van de wervelkolom wordt elke 15 tot 30 seconden verwerkt wanneer de rat wakker is, op zijn vier poten staat en niet beweegt. Als er geen respons wordt waargenomen bij het aanbrengen van een filament, breng dan het volgende filament aan met een hogere kracht in de serie.Als een positieve reactie zoals het buigen van de rug, likken, spiertrekkingen, weggaan van het filament of rotatie wordt waargenomen, gebruik dan het volgende filament met een lagere kracht in de serie. Ga door met de procedure totdat vier metingen zijn verkregen na de eerste gedragsverandering. In deze studie werd de mechanische gevoeligheid tussen CFA en controleratten onderzocht.Bij baseline was de mechanische gevoeligheid vergelijkbaar tussen de groepen. Intramusculaire injectie van CFA in de lumbale spieren veroorzaakte een duidelijke toename van de mechanische gevoeligheid van zeven tot 28 dagen. De controleratten vertoonden daarentegen geen significante verandering.Verder bevestigde histologisch onderzoek de aanwezigheid van chronische inflammatoire veranderingen in de spieren geïnjecteerd met CFA met een duidelijke leukocyteninfiltratie. Gezonde rugspieren van een gecontroleerde rat na intramusculaire injectie van zoutoplossing worden als referentie getoond. De mechanische overgevoeligheid werd waargenomen aan de achterpoot met een standaard Von Frey-test, naast de rugspieren.Het is erg belangrijk om ervoor te zorgen dat de gloeidraad altijd loodrecht op de rug wordt aangebracht en dat het dier niet beweegt bij het aanbrengen. De experimentator moet ook vertrouwen hebben in het onderscheiden van gedragsreacties voor moderne bewegingen.

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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.

Gebruik van de operante Orofacial Pain Assessment Device (OPAD) om veranderingen in Nociceptieve Gedrag Meet

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.

Een protocol van handmatige tests te Sensation en pijn meten bij mensen

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.

De herkenningstest roman-Object voor het onderzoek naar leren en geheugen in muizen

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.

Behandeling van lage rugpijn bij mislukte rugoperatie patiënten met Multicolumn-lead ruggenmerg stimulatie

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.

Het Protocol van de onvoorspelbare chronische milde Stress voor inducerende Anhedonie in muizen

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;

Multimodale signalen voor het analyseren van de pijn reacties op thermische en elektrische 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.

Chronische post-ischemie pijn model voor complexe regionale pijnsyndroom type-I bij ratten

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.

Beoordeling van de geheugenfunctie bij pilocarpine-geïnduceerde epileptische muizen

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.

Chronische stress verschuivingen inspanning-gerelateerde keuze gedrag in een Y-Maze Barrier Taak in muizen

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.

Een verbeterde test en tools voor het meten van mechanische nociceptie in Drosophila-larven

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