This work is funded by the Swiss National Foundation (31_138429) and Synapsis Foundation for the support of research on Alzheimer&#8217;s disease.

Tutte le procedure di animali sono in conformità con la Direttiva UE 2010/63 / UE sulla protezione degli animali utilizzati a fini scientifici e sono approvati dalla locale comitato Animal Care (Cantone di Friburgo, Svizzera). 1. Preparazione degli animali <ol><li> Animali da esperimento <ol><li> Effettuare esperimenti su wild type maschio adulto e topi transgenici (C57BL / 6 di fondo) di 3-5 mesi di età. I tre gruppi di topi corrispondono a controlli di tipo littermate selvatici (gruppo A, Jagged1 flox / flox 19) e due linee del mouse condizionali KO (gruppo B, e C Jagged1ncKO, Jagged1gcKO). </li><li> Topi Camera in condizioni standard di laboratorio in una stanza ventilata, con un ciclo di buio / luce controllata 12 ore e di fornire cibo e acqua ad libitum. </li></ol></li></ol> 2. Setup sperimentale <ol><li> Arena sperimentale <ol><li> Per l&#39;arena sperimentale, utilizzare una gabbia del mouse sterilizzato pulito (36 centimetri di lunghezza x 20,5 centimetri di altezza di larghezza x 13.5 cm) (Figura 1A). </li><li> Assegnare ad ogni mouse per una gabbia numerata con biancheria fresca, alto 3 cm. Se gabbie vengono riutilizzati, come nella prova di sensibilità odore, adottare le seguenti misure per evitare la contaminazione incrociata tra odori e topi. <ol><li> Segnare il lato acqua. </li><li> Pulire le pareti strette delle gabbie con due carte tissue spruzzati con etanolo al 70%, una per ogni lato. </li><li> Accumulano gabbie secondo il genotipo dei topi e memorizzare temporaneamente sotto una cappa laminare. </li></ol></li></ol></li><li> Macchina fotografica <ol><li> Montare una fotocamera su un treppiede personalizzato con l&#39;obiettivo a 58 cm dal fondo della gabbia (Figura 1A). Fissare la posizione del treppiede e la gabbia e delimitare con segni per consentire alla telecamera di essere centrato sulla parte superiore della gabbia. </li><li> Video su 320 pixel x 240 pixel, 15,08 fotogrammi al secondo come file MOV registrare. </li></ol></li><li> Odori <ol><li> Risospendere il scents, quando indicato, nel solvente in cui siano solubili. </li><li> Per il test di preferenza burro di arachidi uso. Risospendere il burro di arachidi in olio di arachidi (10% w / v). </li><li> Per l&#39;uso di test evitamento acido puro 2-metil (2-MB) (98%). </li><li> Per la prova di sensibilità, usare urina femminile dalla stessa colonia mouse e sfondo (C57BL / 6). <ol><li> Per comodità raccogliere le urine 1-2 giorni prima della prova olfattiva. Trattenere e tenere premuto il mouse sotto il cofano con il suo ventre sopra la griglia gabbia. Sotto la griglia gabbia collocare un piatto di plastica petri per raccogliere le gocce di urina. </li><li> Raccogliere l&#39;urina da ciascuna femmina in una provetta da 1,5 ml e miscelare tutti i campioni di urina per normalizzare la variabilità tra animali. Conservare a -20 ° C fino al momento dell&#39;uso. </li><li> Il giorno dell&#39;esperimento, scongelare l&#39;urina ed eseguire 4 diluizioni in acqua bidistillata a un fattore di diluizione di 10 (1:10, 1: 100, 1: 1.000, 1: 10.000). </li></ol></li></ol></li></ol>3. Test olfattiva Nota: in questo protocollo gli odori sono stati volutamente scelti, che sono percepiti come attrattivi forti (burro di arachidi e di urina femminile) o forte repellente (acido 2-MB) 15. È importante effettuare la preferenza e test di sensibilità a piacevoli odori prima della prova di scampo per eliminare la possibilità di eventuali interferenze con il comportamento olfattiva. Tuttavia, per ragioni di semplicità, in questo lavoro, la preferenza e l&#39;elusione di prova saranno entrambi descritti sotto il test di percezione. Ogni sessione comportamentale inizia con una fase di assuefazione. <ol><li> Fase assuefazione <ol><li> Posto l&#39;animale in gabbia assegnato pulito e lasciate esplorare per 5 minuti (Figura 1B). Poiché l&#39;ambiente della gabbia sperimentale è familiare alla gabbia a casa, questo breve periodo di tempo è sufficiente per consentire assuefazione. </li><li> Se il test di sensibilità viene completato in un giorno, effettuare assuefazione sola volta prima della application di alta dell&#39;odore diluito. Se il test di sensibilità viene eseguita in giorni diversi, ogni giorno è necessaria una fase di assuefazione su una nuova gabbia pulita. </li></ol></li><li> Percezione di prova <ol><li> Dopo assuefazione, attivare la fotocamera e subito pipetta 60 ml di profumo gradevole (burro di arachidi) e 60 ml di profumo neutro (acqua di rubinetto) sulle pareti opposte della gabbia a circa 10 cm dal fondo (Figura 1C). </li><li> Lasciate che il mouse esplorare gli odori per 2 minuti (Figura 1D). Successivamente, spegnere la fotocamera. </li><li> A questo punto, procedere con la successiva del mouse a partire dalla fase di assuefazione. Eseguire il test di evitamento esattamente nello stesso modo, applicando 60 ml di odore repellente (acido 2-MB) e 60 ml di acqua. </li></ol></li><li> Test di sensibilità <ol><li> Valutare la soglia attrazione di topi maschi a concentrazioni crescenti di urina femminile nel seguente ordine: 1: 10.000; 1: 1.000; 1: 100; 1:10 e nelle urine puro. </li><li> Dopo assuefazione, esporre ogni topo alla più alta diluizione pipettati dallo sperimentatore come precedentemente descritto in 3.2.1. </li><li> Registrare il comportamento esplorativo di urina contro acqua, entro un arco di tempo 2 minuti su una videocamera. Dopo che tutte le coorti di topi sono testati per la più alta diluizione (1: 10.000), esporre a una maggiore concentrazione di urina, come sopra indicato. </li></ol></li></ol> 4. analisi post-hoc dei dati Nota: Tutti i test comportamentali descritti sono trattati post hoc seguendo le istruzioni di analisi dei dati. <ol><li> Aprire MOV file in ImageJ per i sistemi Windows <ol><li> Installare Quick Time per Java utilizzando le impostazioni personalizzate da http://www.apple.com/quicktime/download. </li><li> Installare il plugin Quick Time dal sito ImageJ (http://rsb.info.nih.gov/ij/plugins/qt-capture.html). </li><li> Importare il QTJava.zip (C: Program Files QuickTime QTSystem) nei extens bibliotecaioni di ImageJ (.ImageJ jre lib ext). </li><li> Copiare anche il QTJava.zip nella cartella plugins e rinominarlo come QTJava.jar. </li><li> Installare i sei script allegati nella cartella macro (ImageJ plugins Macros). </li><li> Aprire ImageJ e compilare ed eseguire il plugin Quick Time, da allora in poi vicino ImageJ. </li><li> Riaprire ImageJ e aprire il file MOV utilizzando File&gt; Importa&gt; utilizzando Quick Time. </li></ol></li><li> Regolazione Video <ol><li> Una volta che il file video viene aperto in ImageJ, tagliare il video per ottenere una costante esplorazione 2 min dal tempo sperimentatore ha pipettato i odoranti nella gabbia (T0). Identificare il telaio corrispondente al T0 e rimuovere i fotogrammi precedenti utilizzando incrementi di 1 (ImageJ immagine Pile ToolsSlice rimozione). Utilizzare lo stesso comando per eliminare tutti i fotogrammi che superano l&#39;esplorazione 2 min. </li><li> Assicurarsi che la gabbia sia centrato e se necessario utilizzare l&#39;Immagine&gt; Trasforma&gt; comando Ruota per allinearlo. </li></ol></li><li> Vedasio Elaborazione Nota: l&#39;elaborazione video è interamente assistita da computer e utilizza i comandi macro che accompagnano questo articolo. <ol><li> Al fine di limitare l&#39;area sulla gabbia di un x 218 pixel dimensioni 127 pixel eseguire la Fase 1 macro dal Plugin&gt; Macro&gt; Esegui comando. Spostare il rettangolo fisso sopra la gabbia (Figura 2, punto 1). </li><li> Raccolto l&#39;area della gabbia sulla regione di interesse (ROI) utilizzando la Fase 2 macro (Figura 2, Passaggio 2). </li><li> Utilizzare la Fase 3 macro per estrarre l&#39;immagine del mouse dallo sfondo assegnando un segnale di soglia, despeckling e filtrando la varianza del segnale. I valori di uscita nel grafico sull&#39;asse Z indicano i valori medi di grigio, corrispondente all&#39;intensità dell&#39;ombra del mouse spostano all&#39;interno della ROI della &quot;camera dell&#39;acqua&quot; durante l&#39;esplorazione 2 min. Copiare i risultati in un foglio di lavoro denominato secondo il ROI in un foglio di calcolo (Figura 2, punto 3). </li><li> Utilizzare il Passo 4 macroestrarre i valori medi grigi del mouse nella ROI &quot;camera di odore&quot;. Copiare i risultati in un foglio di lavoro denominato secondo il ROI nello stesso file foglio di calcolo come a 4.3.3 (Figura 2, punto 4). </li><li> Al fine di limitare ulteriormente l&#39;analisi del movimento del mouse nella ROI &quot;perimetro acqua&quot; utilizzare la macro Passaggio 5. Copiare il risultato sul foglio di lavoro denominato in base al ROI nello stesso file foglio di calcolo come a 4.3.3 (Figura 2, punto 5). </li><li> Per limitare l&#39;analisi del movimento del mouse nella ROI &quot;odore perimetrale&quot; utilizzare la macro Passo 6. Copiare il risultato sul foglio di lavoro denominato in base al file ROI foglio di calcolo come a 4.3.3 (Figura 2, Procedimento 6). </li><li> Elaborare tutti i video e controllare la coerenza del numero di fotogrammi al animali. Qui, registrare tutti gli animali per 1.810 fotogrammi corrispondenti ad una sessione di esplorazione 2 min. </li><li> Per ciascun animale e per ogni ROI sorta telai con media va grigiolues maggiore di 0. Divide il numero di fotogrammi dai valori corrispondenti a 1 secondo e ottenere i secondi trascorsi in ogni ROI. </li></ol></li></ol> 5. Analisi statistica <ol><li> Per ogni prova di verificare l&#39;omogeneità della varianza all&#39;interno gruppi / genotipi con il test di Bartlett con la formula accessibile http://www.real-statistics.com/one-way-analysis-of-variance-anova/homogeneity-variances/. </li><li> Nella attrazione e l&#39;elusione di prova, effettuare confronti tra i tempi trascorsi con acqua contro odore entro un gruppo utilizzando test t di Student non direzionale assumendo varianze uguali o disuguali a seconda dei risultati del test di Bartlett. Confronto i tempi trascorsi con gli odori sottratto dal tempo trascorso con acqua tra le genotipi da ANOVA con test post-hoc di Bonferroni. </li><li> Nel test di sensibilità analizzare i confronti tra il tempo trascorso con l&#39;odore sottratto dal tempo trascorso con water tra i gruppi a una specifica diluizioni di urina da ANOVA con test post-hoc di Bonferroni. Confrontare la sensibilità tra i gruppi a crescenti concentrazioni di odori da 2-Way ANOVA con ripetizioni con test post-hoc di Bonferroni. </li><li> L&#39;interazione tra genotipi e trattamenti nella attrazione e l&#39;elusione di prova sono indagati da 2-way ANOVA con test post-hoc di Bonferroni. </li></ol>

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There is no conflict of interest.

I test proposti in questo protocollo permette di valutare diversi aspetti del comportamento olfattivo innato nei topi: percezione di odori, di discriminazione tra gli odori contro l&#39;acqua e la sensibilità agli odori. Questo protocollo può essere applicato a qualsiasi odore secondo la preferenza e la scala evitamento precedentemente dimostrato 15. Poiché il protocollo si basa sull&#39;attività esplorativa è importante che i topi non presentano deficit motori o ansia che possono influenzare il loro mov...

Olfatto è una delle funzioni sensoriali più sviluppate ed importanti nei mammiferi. Qualsiasi riduzione di valore di attività olfattiva possono influenzare l&#39;assunzione di cibo, comportamento sociale e, nel peggiore dei casi, anche la sopravvivenza. Negli esseri umani, il deterioramento olfattivo è l&#39;età dipendente 1 ed è considerato un forte predittore di disturbi neurologici 2-6. Il test di identificazione olfattiva sviluppato dalla University of Pennsylvania rappresenta attualmente uno dei più utilizzati, non invasive e quantificabili, test diagnostici che possono valutare i primi deficit neurologici 7 e prevedere con elevata probabilità la progressione della demenza 8,9. L&#39;accessibilità del sistema olfattivo e l&#39;importanza dell&#39;olfatto nei roditori, ha scatenato una linea intensa ricerca sulle funzioni meccanismi olfattivi sottostanti 10. Abbiamo precedentemente dimostrato che la perdita di funzione della recept segnalazioneo Notch1 colpisce evitamento olfattiva 11. In questo protocollo si usa topi privi del ligando segnalazione, Jagged1, in neuroni o glia per studiare le prestazioni olfattiva. L&#39;olfatto innata è definito da tre parametri come la percezione, discriminazione tra odori e sensibilità olfattiva 4. Test olfattivo in roditori può essere fatto in una varietà di modi e alcuni studi comportamentali fare uso di olfactometers, che forniscono l&#39;odore all&#39;animale in una specifica concentrazione di vapore e in un lasso di tempo preciso 12 - 14. Tuttavia, questo strumentazione è costoso e può essere disponibile solo in strutture specializzate. Nel nostro lavoro, forniamo un protocollo semplice, veloce e riproducibile test olfattivo, che viene effettuata con profumi volatili. Le prove descritte percezione misura a un attrattivo o un odore repellente e valutare la discriminazione tra il profumo e il 11,15,16 acqua. Utilizzando la stessa messa a punto, we può anche misurare la sensibilità ad un odore a diverse concentrazioni 16,17. L&#39;elaborazione video post-hoc assistita da computer, ispirato all&#39;opera di pagina e colleghi 18, fornisce i risultati imparziali, senza la necessità di accecamento sperimentale e consentendo una sola persona per svolgere l&#39;intero esperimento. Questo protocollo è destinato a fornire un punto di partenza per studiare il comportamento olfattiva nei topi.

<table border="0" cellpadding="0" cellspacing="0" style="border-collapse:
 collapse;width:192pt">
	<colgroup>
		<col span="4" />
	</colgroup>
	<tbody>
		<tr>
			<td class="xl63">Mouse cage</td>
			<td class="xl64">Italplast (Italy)</td>
			<td class="xl64">1144B</td>
			<td class="xl64">36 cm length x 20.5 cm width x 13.5 cm height</td>
		</tr>
		<tr>
			<td class="xl63">Chipped wood bedding</td>
			<td class="xl64">Abedd (Austria)</td>
			<td class="xl64">LTE E-001</td>
			<td class="xl64">3 cm high</td>
		</tr>
		<tr>
			<td class="xl63">Peanut butter</td>
			<td class="xl64">Migros (Swizterland)</td>
			<td class="xl64">NA</td>
			<td align="right" class="xl65">1:10</td>
		</tr>
		<tr>
			<td class="xl63">2-Methylbutyric</td>
			<td class="xl64">Sigma Aldrich (Switzerland)</td>
			<td class="xl64">W269514</td>
			<td class="xl64">Pure</td>
		</tr>
		<tr>
			<td class="xl63">Female Urine from fertile females of same mouse strain</td>
			<td class="xl64">NA</td>
			<td class="xl64">NA</td>
			<td class="xl64">Dilution series</td>
		</tr>
		<tr>
			<td class="xl63">Camera</td>
			<td class="xl64">Olympus (US)</td>
			<td class="xl64">Camedia C-8080</td>
			<td class="xl64">MOV files</td>
		</tr>
		<tr>
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			<td class="xl63">Image J for Windows</td>
			<td class="xl64">NIH (USA)</td>
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simple and computer-assisted olfactory testing for mice

L&#39;olfatto è altamente conservata tra le specie ed è necessario per la riproduzione e la sopravvivenza. Negli esseri umani, l&#39;olfatto è anche uno dei sensi che viene colpito con l&#39;invecchiamento ed è un forte predittore di malattie neurodegenerative. Così, test olfattivo è usato come un metodo diagnostico non invasivo per rilevare deficit neurologici presto. Al fine di comprendere i meccanismi alla base olfattiva suscettibilità rete, ricerca olfattiva nei roditori ha preso slancio negli ultimi dieci anni. Qui, presentiamo un metodo semplice, efficiente e riproducibile test olfattivo tempo di percezione dell&#39;odore innata e sensibilità nei topi senza la necessità di qualsiasi restrizione cibo o acqua prima. I test vengono eseguiti in un ambiente familiare per i topi, richiedono solo i profumi e 2 minuti a sessione di esposizione odorizzante. Viene eseguita l&#39;analisi post-hoc, utilizzando i comandi del computer-assistita in ImageJ e può essere, quindi,, Effettuata dall&#39;inizio alla fine da un ricercatore. Questo protocollo non richiede alcun hardware o configurazione speciale ed è indicato per qualsiasi laboratorio interessati a testare percezione olfattiva e sensibilità.

Il test misura la percezione attrazione di burro di arachidi e di evitare di acido 2-MB. Tre gruppi di topi sono testati e il tempo trascorso in &quot;odore perimetrale&quot; vengono quantificati rispetto all&#39;acqua. Nel test di preferenza, il gruppo di controllo A visualizza preferenza significativa per l&#39;odore rispetto all&#39;acqua (t 8 = 2.52, p &lt;0,05). D&#39;altra parte, il gruppo B non mostra alcuna attrazione significativa al burro di arachidi e passa più tempo con acqua (6 t = 3....

Watch this Scientific Journal Video about Simple and Computer-assisted Olfactory Testing for Mice at JoVE.com

Simple and Computer-assisted Olfactory Testing for Mice

We present a simple and unbiased olfactory test in mice. With this protocol olfactory discrimination, preference, avoidance and sensitivity to a novel odor as compared to water can be assessed in single behavioral sessions. This method is indicated for a single experimenter and analysis is based on computer-assisted video processing.

Semplice e assistita da computer Olfattivo Test per Mice

Olfaction is highly conserved among species and is required for reproduction and survival.
In humans, olfaction is also one of the senses that is affected ...

simple-and-computer-assisted-olfactory-testing-for-mice

Research

JoVE Journal

Behavior

9.8K Views.  University of Fribourg. We present a simple and unbiased olfactory test in mice. With this protocol olfactory discrimination, preference, avoidance and sensitivity to a novel odor as compared to water can be assessed in single behavioral sessions. This method is indicated for a single experimenter and analysis is based on computer-assisted video processing. 

Video: Simple and Computer-assisted Olfactory Testing for Mice

Barnes Maze Testing Strategies with Small and Large Rodent Models

Spatial learning and memory of laboratory rodents is often assessed via navigational ability in mazes, most popular of which are the water and dry-land (Barnes) mazes. Improved performance over sessions or trials is thought to reflect learning and memory of the escape cage/platform location. Considered less stressful than water mazes, the Barnes maze is a relatively simple design of a circular platform top with several holes equally spaced around the perimeter edge. All but one of the holes are false-bottomed or blind-ending, while one leads to an escape cage. Mildly aversive stimuli (e.g.&#160;bright overhead lights) provide motivation to locate the escape cage. Latency to locate the escape cage can be measured during the session; however, additional endpoints typically require video recording. From those video recordings, use of automated tracking software can generate a variety of endpoints that are similar to those produced in water mazes (e.g.&#160;distance traveled, velocity/speed, time spent in the correct quadrant, time spent moving/resting, and confirmation of latency). Type of search strategy (i.e.&#160;random, serial, or direct) can be categorized as well. Barnes maze construction and testing methodologies can differ for small rodents, such as mice, and large rodents, such as rats. For example, while extra-maze cues are effective for rats, smaller wild rodents may require intra-maze cues with a visual barrier around the maze. Appropriate stimuli must be identified which motivate the rodent to locate the escape cage. Both Barnes and water mazes can be time consuming as 4-7 test trials are typically required to detect improved learning and memory performance (e.g.&#160;shorter latencies or path lengths to locate the escape platform or cage) and/or differences between experimental groups. Even so, the Barnes maze is a widely employed behavioral assessment measuring spatial navigational abilities and their potential disruption by genetic, neurobehavioral manipulations, or drug/ toxicant exposure.

The dry-land Barnes maze is widely used to measure spatial navigation ability in response to mildly aversive stimuli. Over consecutive days, performance (e.g.&#160;latency to locate escape cage) of control subjects improves, indicative of normal learning and memory. Differences between rats and mice necessitate apparatus and methodology changes that are detailed here.

Barnes strategie di sperimentazione Maze con piccole e grandi modelli di roditori

Spatial learning and memory of laboratory rodents is often assessed via navigational ability in mazes, most popular of which are the water and dry-land ...

Ricerca

Comportamento

Testing for Odor Discrimination and Habituation in Mice

This video demonstrates a technique to establish the presence of a normally functioning olfactory system in a mouse. The test helps determine whether the mouse can discriminate between non-social odors and social odors, whether the mouse habituates to a repeatedly presented odor, and whether the mouse demonstrates dishabituation when presented with a novel odor. Since many social behavior tests measure the experimental animal&#8217;s response to a familiar or novel mouse, false positives can be avoided by establishing that the animals can detect and discriminate between social odors. There are similar considerations in learning tests such as fear conditioning that use odor to create a novel environment or olfactory cues as an associative stimulus. Deficits in the olfactory system would impair the ability to distinguish between contexts and to form an association with an olfactory cue during fear conditioning. 
In the odor habitation/dishabituation test, the mouse is repeatedly presented with several odors. Each odor is presented three times for two minutes. The investigator records the sniffing time directed towards the odor as the measurement of olfactory responsiveness. A typical mouse shows a decrease in response to the odor over repeated presentations (habituation). The experimenter then presents a novel odor that elicits increased sniffing towards the new odor (dishabituation). After repeated presentation of the novel odor the animal again shows habituation. This protocol involves the presentation of water, two or more non-social odors, and two social odors. In addition to reducing experimental confounds, this test can provide information on the function of the olfactory systems of new knockout, knock-in, and conditional knockout mouse lines.

This manuscript describes a protocol to examine the olfactory system of rodents. The olfactory habituation/dishabituation test will allow investigators to determine whether a mouse habituates to a repeatedly presented odor and whether the mouse demonstrates dishabituation when presented a novel odor.

Test per Odore Discriminazione e assuefazione nei topi

This video demonstrates a technique to establish the presence of a normally functioning olfactory system in a mouse. The test helps determine whether the ...

Morris Water Maze Test: Optimization for Mouse Strain and Testing Environment

The Morris water maze (MWM) is a commonly used task to assess hippocampal-dependent spatial learning and memory in transgenic mouse models of disease, including neurocognitive disorders such as Alzheimer&#8217;s disease. However, the background strain of the mouse model used can have a substantial effect on the observed behavioral phenotype, with some strains exhibiting superior learning ability relative to others. To ensure differences between transgene negative and transgene positive mice can be detected, identification of a training procedure sensitive to the background strain is essential. Failure to tailor the MWM protocol to the background strain of the mouse model may lead to under- or over- training, thereby masking group differences in probe trials. Here, a MWM protocol tailored for use with the F1 FVB/N x 129S6 background is described. This is a frequently used background strain to study the age-dependent effects of mutant P301L tau (rTg(TauP301L)4510 mice) on the memory deficits associated with Alzheimer&#8217;s disease. Also described is a strategy to re-optimize, as dictated by the particular testing environment utilized.

This manuscript describes a Morris water maze (MWM) protocol tailored for use with a commonly used mouse model of Alzheimer's disease. The MWM is widely used in transgenic mouse models. Implementation of a procedure sensitive to the background strain of the mouse model is essential for detecting group differences.

Morris water maze Test: Ottimizzazione per ceppo di topi e Ambiente di prova

 The Morris water maze (MWM) is a commonly used task to assess hippocampal-dependent spatial learning and memory in transgenic mouse models of disease, ...

The Treadmill Fatigue Test: A Simple, High-throughput Assay of Fatigue-like Behavior for the Mouse

Fatigue is a prominent symptom in many diseases and disorders and reduces quality of life for many people. The lack of clear pathogenesis and failure of current interventions to adequately treat fatigue in all patients leaves a need for new treatment options. Despite the therapeutic need and importance of preclinical research in helping identify promising novel treatments, few preclinical assays of fatigue are available. Moreover, the most common preclinical assay used to assess fatigue-like behavior, voluntary wheel running, is not suitable for use with some strains of mice, may not be sensitive to drugs that reduce fatigue, and has relatively low throughput. The current protocol describes a novel, non-voluntary preclinical assay of fatigue-like behavior, the treadmill fatigue test, and provides evidence of its efficacy in detecting fatigue-like behavior in mice treated with a chemotherapy drug known to cause fatigue in humans and fatigue-like behavior in animals. This assay may be a beneficial alternative to wheel running, as fatigue-like behavior and potential interventions can be assessed in a greater number of mice over a shorter time frame, thus permitting faster discovery of new therapeutic options.

Fatigue is a common, undertreated and frequently poorly-understood symptom in many diseases and disorders. New preclinical assays of fatigue may help to improve current understanding and future treatment of fatigue. To that end, the current protocol provides a novel means of measuring fatigue-like behavior in the mouse.

Il tapis roulant fatica di prova: un test semplice, high-throughput di stanchezza simile comportamento per il mouse

Fatigue is a prominent symptom in many diseases and disorders and reduces quality of life for many people. The lack of clear pathogenesis and failure of ...

Task Interruption and Resumption Paradigm for Testing the Activation and Pursuit of an Abstract Thinking Goal

This protocol is based on the task interruption and resumption paradigm, the premise of which is that active goals lead to persistent behavior and thus a higher resumption rate after a period of delay or interruption. The task interruption and resumption protocol described in this research is tailored to test the activation of cognitive goals (e.g., a goal to think more abstractly). Cognitive goals may be pursued even during the interruption period; thus, to prevent this, the protocol involves cognitive distraction. The protocol consists of several stages. Specifically, the initial stage includes the goal activation process, where the treatment (versus control) condition receives a manipulation to activate the cognitive goal being tested by the researcher. In the next stage, participants are presented with the introduction of a task that is perceived to either satisfy or not satisfy the cognitive goal of interest. Importantly, this task is interrupted a few seconds after it begins. The task interruption forces a delay period and introduces a cognitive distraction to prevent the automatic pursuit and fulfillment of the cognitive goal. After the interruption period, participants are given a choice between resuming the interrupted task and abandoning the interrupted task to complete an alternative task instead. Among participants whose cognitive goals had been activated at the earlier stage, the task resumption rate should be higher if the task was perceived as an opportunity to satisfy (versus not satisfy) the goal. Such a finding would provide empirical evidence that the cognitive goal has been activated and pursued. In previous research, this protocol has been used to test whether causal uncertainty activates an abstract thinking goal. Adapting the protocol to test the activation of other cognitive goals is also discussed.

This protocol was designed to test the activation and pursuit of cognitive goals (e.g., an abstract thinking goal) using the task interruption and resumption paradigm. The protocol is suitable for cognitive goals that are automatically pursued once activated, as the distraction procedure prevents goal pursuit during the interruption period.

Task Interruzione e paradigma Ripresa per il test l&#39;attivazione e perseguimento di un obiettivo astratto Thinking

This protocol is based on the task interruption and resumption paradigm, the premise of which is that active goals lead to persistent behavior and thus a ...

A Simple and Low-cost Assay for Measuring Ambulation in Mouse Models of Muscular Dystrophy

Measuring functional outcomes in the treatment of muscular dystrophy is an essential aspect of preclinical testing. The assessment of voluntary ambulation in mouse models is a non-invasive and reproducible activity assay that is directly analogous to measures of patient ambulation such as the 6-minute walk test and related mobility scores. Many common methods for testing mouse ambulation speed and distance are based on the open field test, where an animal&#39;s free movement within an arena is measured over time. One major downside to this approach is that commercial software and equipment for high-resolution motion tracking is expensive and may require transferring mice to specialized facilities for testing. Here, we describe a low-cost, video-based system for measuring mouse ambulation that utilizes free and open-source software. Using this protocol, we demonstrate that voluntary ambulation in the dystrophin-null mdx mouse model for Duchenne muscular dystrophy (DMD) is decreased relative to wild-type mouse activity. In mdx mice expressing the utrophin transgene, these activity deficits are not observed and the total distance traveled is indistinguishable from wild-type mice. This method is effective for measuring changes in voluntary ambulation associated with dystrophic pathology, and provides a versatile platform that can be readily adapted to diverse research settings.

This protocol describes a flexible, low-cost system for measuring mouse ambulation in an open field activity assay. We show that a 6-minute ambulation assay based on this system detects a decrease in voluntary movement in mdx mice, and accurately distinguishes improvement in a muscle-specific rescue of these animals.

Un'analisi semplice e a basso costo per la misurazione della deambulazione in modelli murini di distrofia muscolare

Measuring functional outcomes in the treatment of muscular dystrophy is an essential aspect of preclinical testing. The assessment of voluntary ambulation ...

A Simple Non-invasive Method for Temporary Knockdown of Upper Limb Proprioception

Proprioception may be the least well measured of all contributors to the neural control of movement. New precise, reliable measures of proprioception are needed for clinical diagnosis of impairment, and to measure outcomes of proprioceptive training. The purpose of this simple, non-invasive method is to temporarily knockdown upper limb proprioception in healthy adults, to an extent that would be useful in the development and testing of upper limb proprioception measures. Knockdown models have two main advantages over studying humans with impaired proprioception: participant availability and the ability to control the extent of impairment across participants. Current published methods of temporary proprioception knockdown of the upper limb, such as ischemic nerve blocks and cryotherapy, are invasive, impractical, or uncomfortable for the participant. Here, vibration over the ulnar groove was used to reduce upper limb proprioception. High frequency vibration may reduce proprioceptive acuity by inhibiting pacinian corpuscle-induced input. The effect of vibration used in this protocol was confirmed using two quantitative measures. This method was simple to administer, comfortable for participants, and practical.

The goal of this protocol is to demonstrate a practical method to temporarily interfere with proprioception in the upper limb of healthy humans.

Un metodo semplice e Non invasivo per atterramento temporaneo di propriocezione degli arti superiori

Proprioception may be the least well measured of all contributors to the neural control of movement. New precise, reliable measures of proprioception are ...

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.

Attiva e passiva docilità separatamente nei topi di misura

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 ...

Computerized Adaptive Testing System of Functional Assessment of Stroke

The computerized adaptive testing system of the functional assessment of stroke (CAT-FAS) can simultaneously assess four functions (motor functions of the upper and lower extremities, postural control, and basic activities of daily living) with sufficient reliability and administrative efficiency. CAT, a modern measurement method, aims to provide a reliable estimate of the examinee's level of function rapidly. CAT administers only a few items whose item difficulties match an examinee's level of function and, thus, the administered items of CAT can provide sufficient information to reliably estimate the examinee's level of function in a short time. The CAT-FAS was developed through four steps: (1) determining the item bank, (2) determining the stopping rules, (3) validating the CAT-FAS, and (4) establishing a platform of online administration. The results of this study indicate that the CAT-FAS has sufficient administrative efficiency (average number of items = 8.5) and reliability (group-level Rasch reliability: 0.88 - 0.93; individual-level Rasch reliability: &#8805;70% of patients had Rasch reliability score &#8805;0.90) to simultaneously assess four functions in patients with stroke. In addition, because the CAT-FAS is a computer-based test, the CAT-FAS has three additional advantages: the automatic calculation of scores, the immediate storage of data, and the easy exporting of data. These advantages of the CAT-FAS will be beneficial to data management for clinicians and researchers.

Here, we present a protocol to develop the computerized adaptive testing system of the functional assessment of stroke (CAT-FAS). The CAT-FAS can simultaneously assess four functions (two motor functions [upper and lower extremities], postural control, and basic activities of daily living) with sufficient reliability and administrative efficiency.

Sistema computerizzato di test adattivo della valutazione funzionale del colpo

The computerized adaptive testing system of the functional assessment of stroke (CAT-FAS) can simultaneously assess four functions (motor functions of the ...

Olfactory Context Dependent Memory: Direct Presentation of Odorants

Information is retrieved more effectively when the retrieval occurs in the same context as that in which the information was first encoded. This is termed context dependent memory (CDM). One category of cues that have been shown to effectively produce CDM effects are odors. However, it is unclear what the boundary conditions of these CDM effects are. In particular, given that olfaction has been called an implicit sense, it is possible that odors are only effective mnemonic cues when they are presented in the background. This assertion seems even more likely given that previous research has shown odors to be poor cues during paired associate memory tests, where odors are in the focus of attention as mnemonic cues for other information. In order to determine whether odors are only effective contextual mnemonic cues when presented outside the central focus of an observer, an olfactory CDM experiment was performed in which odorants were presented directly, rather than ambiently. Direct presentation was accomplished with the aid of an olfactometer. The olfactometer not only allows for direct presentation of odorants, but provides other methodological benefits, including the allowance of trial by trial manipulations of odorant presentations and, relatedly, time-specific releases of odorants. The presence of the same odor during both encoding and retrieval enhanced memory performance, regardless of whether the odor was presented ambiently or directly. This finding can serve as a basis for future olfactory CDM research which can utilize the benefits of direct presentation.

The use of an olfactometer for directly presenting odorants opens exciting opportunities for researchers of olfactory memory. The current paper discusses issues related to this methodology as related to a previously published experiment on olfactory context dependent memory.

Memoria dipendente dal contesto olfattivo: Presentazione diretta di odorizzazione

Information is retrieved more effectively when the retrieval occurs in the same context as that in which the information was first encoded. This is termed ...