This study was supported by the American Heart Association (IRG 14330015, PRE 16850005, PRE 20380386, EIA 0740129N), and the National Institutes of Health (RR 2865AR; P20 RR 016477).

Tutte le procedure descritte di seguito sono stati esaminati e approvati dalla cura degli animali e Usa Comitato Istituzionale al West Virginia University Health Sciences Center. 1. Selezione di un modello animale NOTA: Le specie più comunemente utilizzati nel modello UCMS includono Sprague Dawley, e ratti Wistar e la BALB / CJ del mouse; altri modelli murini hanno dimostrato l&#39;efficacia limitata (DBA / 2, C57BL / 6). Tuttavia, un&#39;attenta considerazione dovrebbe essere data al particolare mouse o ceppo di ratti di scelta, come il background genetico e suscettibilità a stress ambientali variano ampiamente tra le specie e hanno dimostrato di essere fattori critici che influenzano i risultati del 11,14,15 studio, 19. <ol><li> Utilizzare gli animali di età appropriata (ad esempio, se l&#39;endpoint desiderato deve essere valutata in un 17 settimane vecchio topo, poi i ratti devono iniziare il protocollo UCMS a ~ 9 settimane di età) in quanto questo protocollo prende almst 7-9 settimane per indurre completamente sintomi depressivi. </li><li> Animali Singolo-house sottoposti UCMS in gabbie e strutture IACUC approvati, con il cibo e il chow di serie ad libitum disponibile (salvo diversa prescrizione specifico protocollo). </li><li> Fare particolare attenzione a garantire controlli sono alloggiati in una stanza tranquilla con attività minima per ridurre gli effetti di eventuali fattori di stress ambientali sconosciuti. </li></ol> 2. Imprevedibile cronica lieve stress protocollo NOTA: Gli animali sottoposti UCMS vengono trasportati ogni giorno (&gt; 5 giorni alla settimana) per una camera pulita utilizzata per le manipolazioni UCMs. Gli animali sono esposti a 1 o 2 dei fattori di stress sotto elencati ogni giorno. Fattori di stress sono eseguite su un programma randomizzato, e ogni sforzo è somministrato per un minimo di 3-4 ore (vedere Tabella 1). <ol><li> Inumidire la biancheria da letto versando 10-20 oncia acqua di pulito in ogni gabbia standard. Regolare la quantità di acqua specialmente per la misura dellagabbia per smorzare completamente la biancheria da letto, ma non provocare il pool di acqua. Conservati il ​​animali in gabbia umida per 3-4 ore. </li><li> Togliere biancheria da letto da ogni gabbia per 3-4 ore, dopo di che gli animali sono la transizione in un altro fattore di stress che richiede una gabbia vuota (bagno poco profondo, gabbia tilt) o posto in una gabbia pulita con biancheria fresca. </li><li> Gabbie Tilt a circa 45 ° (senza biancheria da letto) per 3-4 ore. Gabbie possono essere inclinati con un oggetto solido che rimarrà in luogo come l&#39;animale si muove nella gabbia. </li><li> Alter normali cicli giorno / notte 12 ore di luce / 12 ore in successioni di 30 periodi minimo della durata di 8 ore. Dopo, gli animali tornare al normale ciclo. </li><li> Trasferire ciascun animale dalla gabbia sede che della gabbia di un animale vicina che è stato rimosso per 3 ore. </li><li> Remove tutte le lenzuola da ogni gabbia e aggiungere acqua fino ad una profondità pari a ~ 0,25 pollici per topi o ~ 0.5 pollici per i ratti per circa 4 ore. Usare acqua che è più calda di RT (~ 30 ° C) per ridurre al minimo hypothermpotenziale ia. Brevemente animali secco con un asciugamano morbido a conclusione dell&#39;esposizione acqua prima del posizionamento in gabbie pulite. </li><li> Esporre animali predatore odori / suoni con l&#39;aggiunta di ciuffi di campioni distribuiti in modo casuale di pelliccia o nelle urine (10-20 ml) da una specie di predatori naturali nella gabbia per 3-4 ore. In alternativa, giocare a ringhiare o rumori predatori di specie di predatori naturali in prossimità di gabbie per 3 ore. </li><li> Al termine di ciascun periodo di stress quotidiano, mettere tutti gli animali in gabbie pulite e restituirli alla struttura alloggiativa. </li></ol> 3. Monitoraggio animali / Grooming punteggi <ol><li> Controllare ogni animale giornaliera per lo sviluppo di ferite o piaghe che possono richiedere trattamento veterinario. Gli animali che si sviluppano lesioni lievi possono continuare nel protocollo UCMS sul consenso del veterinario. <ol><li> Due investigatori addestrati nella valutazione cappotto toelettatura ispezionare ogni animale e assegnano un punteggio cappotto settimanale su una scala 0-1. Questo punteggio è scoraggiareminato assegnando punteggi individuali (0- pulito, 1-sporco) a ciascuno di testa 8 regioni: corpo, collo, cappotto dorsale, cappotto ventrale, coda, arti anteriori, arti posteriori, e della regione genitale. Gli stessi ricercatori sono per eseguire questa operazione in tutto il protocollo UCMS per mantenere la coerenza in l&#39;assegnazione di punteggi cappotto. </li><li> Ogni animale che vive una forte riduzione del peso corporeo (cioè,&gt; 15%) devono essere segnalati al personale veterinario e deve essere rimosso dal protocollo. </li></ol></li></ol> 4. comportamentale Prove e misure di outcome NOTA: Diversi metodi di valutazione può essere utilizzato per quantificare i cambiamenti comportamentali del protocollo UCMS nei roditori. <ol><li> Cumulative Score Coat Grooming. <ol><li> Come descritto in precedenza, un punteggio cappotto governare settimanale viene assegnato per ciascun animale. </li></ol></li><li> Saccarosio Splash prova: <ol><li> Posto l&#39;animale in una gabbia pulita foderato con un batuffolo di cotone o un asciugamano per evitare biancheria da lettodi aderire al suo cappotto. Spruzzare una soluzione di saccarosio al 10% sulla testa e dorsale mano di ogni animale e registrare il comportamento complessivo governare per 5 min. </li><li> Governare l&#39;attività (definito come la pulizia del pelo leccando o graffi) si misura con latenza (tempo di inattività tra il primo spruzzo e l&#39;avvio di grooming) e frequenza (numero di volte che governare una particolare parte del corpo). </li></ol></li><li> Saccarosio Preferenza Test: <ol><li> Abituare animali bere 1% di soluzione di saccarosio (w / v) 72 hr prima della prova e quindi fornire esposizione a due bottiglie (soluzione di saccarosio al 1% contro acqua di rubinetto in diverse bottiglie). Dopo assuefazione, fornire animali con ad libitum l&#39;accesso a una soluzione e rubinetto di saccarosio per 3 ore. Dopo circa 3 ore di registrazione dei volumi di consumo di soluzione di saccarosio e acqua di rubinetto e calcolare la preferenza di saccarosio come: [(consumo saccarosio) / (consumo di acqua + Consumo saccarosio)] × 100. </li></ol></li><li> Tail sospensione di prova (Mouse Solo): <ol><li> Sutrascorrere topi per la coda da un asta stabile (20 cm dal pavimento) con del nastro adesivo (2 cm dalla punta della coda). Tempo di immobilità Record su 5 min. I topi sono considerati immobili solo quando si appendono passivamente e completamente immobile. </li></ol></li><li> Il Swim prova forzata: <ol><li> Posizionare gli animali in un contenitore (~ 15-20 cm di diametro per i topi; ~ 55-60 cm di diametro per i ratti) riempiti con acqua ad una temperatura di 26 ± 0,5 ° C. Registrare la latenza per il primo periodo della mobilità e il tempo trascorso immobile durante il periodo di prova di 5 min. Alla fine del test, posizionare l&#39;animale bagnato in una gabbia tenuta su una piastra elettrica (30-35 ° C) con biancheria normale coperto da un foglio di carta assorbente fino a secco. </li></ol></li><li> Più elevata labirinto: <ol><li> Consentire ad ogni animale per esplorare il labirinto liberamente per 5 min. Registra voci totali in ogni braccio, e il tempo totale trascorso in ogni braccio, secondo un criterio a priori 20. </li></ol></li><li> Locomotor attività: <ol><li> Livelli di attività record, definiti come movimento complessivo, movimenti fini, movimenti allevamento, e l&#39;inattività, sia acuta (30 min) e / o cronica (12 hr) periodi di tempo. Farlo utilizzando la gabbia casa o di un nuovo ambiente, a seconda dell&#39;ambiente individuo (anche se la coerenza dovrebbe essere mantenuta) in tempo reale da un osservatore o post-hoc per la riproduzione e l&#39;analisi di filmati della telecamera. </li></ol></li></ol> 5. Espansione / Variazione del protocollo UCMS <ol><li> Amministrare appropriati agenti terapeutici o le sfide di studiare la loro capacità di smussare o lo sviluppo dei risultati UCMS-indotti (se avviato precoce) o per invertire la gravità dei risultati UCMS indotte stabiliti (se iniziata in ritardo). Questo dipende dal singolo laboratorio o protocollo 11. </li></ol> 6. Valutazioni Risultato finale <ol><li> A seguito della conclusione di protocolli UCMS e test comportamentali, utilizzare animali in seguito esperiiments in modo che affronti le ipotesi specifiche, obiettivi o obiettivi del singolo laboratorio. NOTA: Come esempi, questo può includere le risposte cardiovascolari, infiammatorie o endocrine allo stress cronico. In alternativa, gli interventi terapeutici possono essere introdotti a questo punto per valutare l&#39;efficacia e basi meccanicistici. </li></ol>

<ol>
	<li>Lett, H. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Depression+as+a+risk+factor+for+coronary+artery+disease:+evidence,+mechanisms,+and+treatment.">Depression as a risk factor for coronary artery disease: evidence, mechanisms, and treatment.</a> Psychosom Med. 66 (3), 305-315 (2004).</li><li>Plante, G. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Depression+and+cardiovascular+disease:+a+reciprocal+relationship.">Depression and cardiovascular disease: a reciprocal relationship.</a> Metabolism. 54, 45-48 (2005).</li><li>Pizzi, C., Manzoli, L., Mancini, S., Bedetti, G., Fontana, F., Costa, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Autonomic+nervous+system,+inflammation+and+preclinical+carotid+atherosclerosis+in+depressed+subjects+with+coronary+risk+factors.">Autonomic nervous system, inflammation and preclinical carotid atherosclerosis in depressed subjects with coronary risk factors.</a> Atherosclerosis. 212 (1), 292-298 (2010).</li><li>Barden, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Implication+of+the+hypothalamic-pituitary-adrenal+axis+in+the+physiopathology+of+depression.">Implication of the hypothalamic-pituitary-adrenal axis in the physiopathology of depression.</a> J Psychiatry Neurosci. 29 (3), 185-193 (2004).</li><li>Bowman, R. E., Beck, K. D., Luine, V. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+stress+effects+on+memory:+sex+differences+in+performance+and+monoaminergic+activity.">Chronic stress effects on memory: sex differences in performance and monoaminergic activity.</a> Horm Behav. 43, 48-59 (2003).</li><li>Baune, B. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+relationship+between+subtypes+of+depression+and+cardiovascular+disease:+a+systematic+review+of+biological+models.">The relationship between subtypes of depression and cardiovascular disease: a systematic review of biological models.</a> Transl Psychiatry. 2, e92 (2012).</li><li>O'Leary, O. F., Cryan, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Towards+translational+rodent+models+of+depression.">Towards translational rodent models of depression.</a> Cell Tissue Res. 354 (1), 141-153 (2013).</li><li>Mineur, Y. S., Belzung, C., Crusio, W. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+unpredictable+chronic+mild+stress+on+anxiety+and+depression-like+behavior+in+mice.">Effects of unpredictable chronic mild stress on anxiety and depression-like behavior in mice.</a> Behav Brain Res. 175 (1), 43-50 (2006).</li><li>Yalcin, I., Belzung, I., Surget, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+strain+differences+in+the+unpredictable+chronic+mild+stress:+a+four-antidepressant+survey.">Mouse strain differences in the unpredictable chronic mild stress: a four-antidepressant survey.</a> Behav Brain Res. 193 (1), 140-143 (2008).</li><li>d'Audiffret, A. C., Frisbee, S. J., Stapleton, P. A., Goodwill, A. G., Isingrini, E., Frisbee, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Depressive+behavior+and+vascular+dysfunction:+a+link+between+clinical+depression+and+vascular+disease.">Depressive behavior and vascular dysfunction: a link between clinical depression and vascular disease.</a> J Appl Physiol. 108 (5), 1041-1051 (2010).</li><li>Stanley, S. C., Brooks, S. D., Butcher, J. T., d'Audiffret, A. C., Frisbee, S. J., Frisbee, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protective+effect+of+sex+on+chronic+stress-+and+depressive+behavior-induced+vascular+dysfunction+in+BALB/cJ+mice.">Protective effect of sex on chronic stress- and depressive behavior-induced vascular dysfunction in BALB/cJ mice.</a> J Appl Physiol. 117 (9), 959-970 (2014).</li><li>Ibarguen-Vargas, Y., Surget, A., Touma, C., Palme, R., Belzung, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multifaceted+strain-specific+effects+in+a+mouse+model+of+depression+and+of+antidepressant+reversal.">Multifaceted strain-specific effects in a mouse model of depression and of antidepressant reversal.</a> Psychoneuroendocrinology. 33 (10), 1357-1368 (2008).</li><li>Dalla, C., Pitychoutis, P. M., Kokras, N., Papadopoulou-Daifoti, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sex+differences+in+animal+models+of+depression+and+antidepressant+response.">Sex differences in animal models of depression and antidepressant response.</a> Basic Clin Pharmacol Toxicol. 106 (3), 226-233 (2010).</li><li>Mutlu, O., Gumuslu, E., Ulak, G., Celikyurt, I. K., Kokturk, S., K&#305;r, H. M., Akar, F., Erden, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+fluoxetine,+tianeptine+and+olanzapine+on+unpredictable+chronic+mild+stress-induced+depression-like+behavior+in+mice.">Effects of fluoxetine, tianeptine and olanzapine on unpredictable chronic mild stress-induced depression-like behavior in mice.</a> Life Sci. 91 (25-26), 1252-1262 (2012).</li><li>Gumuslu, E., Mutlu, O., Sunnetci, D., Ulak, G., Celikyurt, I. K., Cine, N., Akar, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effects+of+tianeptine,+olanzapine+and+fluoxetine+on+the+cognitive+behaviors+of+unpredictable+chronic+mild+stress-exposed+mice.">The effects of tianeptine, olanzapine and fluoxetine on the cognitive behaviors of unpredictable chronic mild stress-exposed mice.</a> Drug Res (Stuttg). 63 (10), 532-539 (2013).</li><li>Isingrini, E., Belzung, C., Freslon, J. L., Machet, M. C., Camus, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fluoxetine+effect+on+aortic+nitric+oxide-dependent+vasorelaxation+in+the+unpredictable+chronic+mild+stress+model+of+depression+in+mice.">Fluoxetine effect on aortic nitric oxide-dependent vasorelaxation in the unpredictable chronic mild stress model of depression in mice.</a> Psychosom Med. 74 (1), 63-72 (2012).</li><li>Ripoll, N., David, D. J., Dailly, E., Hasco&#235;t, M., Bourin, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antidepressant-like+effects+in+various+mice+strains+in+the+tail+suspension+test.">Antidepressant-like effects in various mice strains in the tail suspension test.</a> Behav Brain Res. 143, 193-200 (2003).</li><li>Komada, M., Takao, K., Miyakawa, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Elevated+plus+maze+for+mice.">Elevated plus maze for mice.</a> J Vis Exp. (22), 1088 (2008).</li><li>Golbidi, S., Frisbee, J. C., Laher, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+stress+impacts+the+cardiovascular+system:+animal+models+and+clinical+outcomes.">Chronic stress impacts the cardiovascular system: animal models and clinical outcomes.</a> Am. J. Physiol. Heart Circ. Physiol. , (2015).</li></ol>

The authors have nothing to disclose.

Extensive clinical and epidemiological evidence has indicated that chronic stress is one of the most potent precipitating factors for depression. Repeated exposure to stressors may potentiate individual vulnerability to depression and other neuropsychiatric disorders as a consequence of the psychological and physical demands within the body that accumulate over time. The use of validated preclinical animal models offers a valuable translational tool for studying depression and comorbid diseases. Firstly, an ideal animal ...

La malattia mentale depressiva è un disordine neurologico complesso che è attualmente riconosciuta come una delle principali cause di disabilità e malattia onere in tutto il mondo. Il NIMH riferisce che circa il 12% degli americani soffre di depressione clinica, con il doppio delle donne colpite contro uomini 1. Solo negli Stati Uniti, la depressione rappresenta per miliardi di dollari i costi sanitari diretti e circa $ 193.000.000.000 più nei costi indiretti (abbassato guadagni e perdite di produttività) 2. I sintomi della depressione includono anedonia, i cambiamenti nei cicli di peso e di sonno, ridotta attività fisica e l&#39;igiene personale, sentimenti di disperazione o di colpa, e / o pensieri ricorrenti di morte o di suicidio. Durante l&#39;ultimo decennio, epidemiologici e studi clinici hanno indicato che la depressione è un fattore di rischio indipendente per la malattia cardiovascolare (CVD) morbilità e mortalità a 3, ed è predittivo di più grave prognosi di patologie cardiovascolari, tra cui untherosclerosis, ipertensione, infarto del miocardio, e malattia coronarica, indipendentemente dalla precedente storia di malattia cardiovascolare conclamata 4. Nonostante la crescente prevalenza e l&#39;impatto negativo sulla salute pubblica di depressione, l&#39;eziologia e la relativa fisiopatologia di questa malattia è poco conosciuta e l&#39;eterogeneità della malattia a causa di vari fattori (ad esempio,, biologici, e componenti ambientali genetiche) ha reso diagnosi clinica difficile definire. Le prove indicano che lo stress psicologico irrisolvibile è un fattore importante che contribuisce per lo sviluppo di malattie depressive e può anche essere un potente fattore patogeno che collega la depressione e malattie cardiovascolari, in parte a causa di interruzioni e disregolazione dell&#39;asse ipotalamo-ipofisi-surrene (HPA) 5,6. Disfunzione dell&#39;asse HPA è un meccanismo importante che è stato collegato ai cambiamenti comportamentali e fisiologiche osservate in depressione e lo sviluppo di diversi fattori di rischio CVD, compresidislipidemia, obesità, diabete e 7. Molteplici modelli preclinici di depressione sono stati sviluppati nel tentativo di replicare il meccanismo di alterazione dell&#39;attività HPA asse caratteristico della depressione clinica; tali modelli forniscono un mezzo convalidati per indagare i cambiamenti comportamentali, neurologici e fisiologici associati con lo stress cronico e acuto negli animali. La validità di un modello animale della malattia si basa sulla rilevanza del l&#39;eziologia e la progressione del modello di design e la sua capacità di ricapitolare anatomica, neurofisiologici, e le caratteristiche comportamentali osservati in malattie umane. Inoltre, le risposte preclinici ai trattamenti (come SSRI) dovrebbero portare a risultati simili a quelli osservati in ambito clinico. Diversi modelli animali di depressione indotta da stress sono attualmente utilizzati nella ricerca, come ad esempio impotenza appreso, lo stress primi anni di vita, e lo stress sconfitta sociale. Tuttavia, ciascuno di questi modelli presenta svantaggi intrinseciche riducono la loro efficacia traslazionale 8. Negli ultimi decenni, il protocollo Imprevedibile cronica stress lieve (UCMS) è emerso come uno dei modelli più translationally rilevanti per lo studio della fisiopatologia della depressione nei roditori 9. Questo modello è basato sul concetto fondamentale che l&#39;esposizione cronica a fattori di stress distrugge i sistemi di risposta allo stress e alla fine porta allo sviluppo di disturbi depressivi. Durante il protocollo UCMS, gli animali sono esposti a una serie di fattori di stress randomizzato ambientali e sociali miti su base giornaliera. Un fattore cruciale che aumenta la rilevanza di questo modello a situazioni umane risiede nell&#39;elevato livello di imprevedibilità e incontrollabilità dei fattori di stress, come pure il momento in cui sono introdotti. Inoltre, il protocollo UCMS utilizza solo stress lievi, invece di basarsi su primi anni di vita o di stimoli fisici aggressivi. In un periodo di esposizione UCMS, comportamenti depressivi sviluppano e sono paragonabili a clinsintomi iCal, tra cui diminuzione dell risposta alle ricompense (anedonia), cambiamenti di attività fisica e il comportamento investigativo (impotenza e disperazione), il deterioramento dello stato di cappotto e alterato l&#39;attività sessuale 10. Quasi tutti i sintomi della depressione dimostrabili sono stati riportati utilizzando questo modello, e gli studi hanno dimostrato che questi comportamenti persistono per diverse settimane dopo la cessazione del dolore. Inoltre, questi comportamenti depressivi UCMS-indotte possono essere gradualmente venerati da cronica, ma non acuta, il trattamento con particolari antidepressivi, suggerendo effetti neurologici simili di miglioramento terapeutico che rispecchia da vicino l&#39;azione e l&#39;efficacia clinica variabile di questi agenti negli esseri umani 11,14- 18. Qui riportiamo una descrizione dettagliata del protocollo UCMS e descriviamo risultati tipici comportamentali e vascolari in topi.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Temporary animal cages with lids</td>
			<td>n/a</td>
			<td>n/a&#160;</td>
			<td>Provided by your animal care facility</td>
		</tr>
		<tr>
			<td>Rodent bedding</td>
			<td>n/a</td>
			<td>n/a&#160;</td>
			<td>Provided by your animal care facility</td>
		</tr>
		<tr>
			<td>Predator fur or urine</td>
			<td>n/a</td>
			<td>n/a&#160;</td>
			<td>Provided by your animal care facility</td>
		</tr>
		<tr>
			<td>Rodent cage drinking bottles</td>
			<td>n/a</td>
			<td>n/a&#160;</td>
			<td>Provided by your animal care facility</td>
		</tr>
		<tr>
			<td>1000 ml graduated cylinder</td>
			<td>variable</td>
			<td>variable</td>
			<td>This is optional.&#160; Any container from which a known quantiity of water can be poured will be appropriate</td>
		</tr>
		<tr>
			<td>Wooden blocks cut from 2x4</td>
			<td>variable</td>
			<td>variable</td>
			<td>8 inch sections are cut to facilitate cage tilt procedures</td>
		</tr>
		<tr>
			<td>Soft paper towels</td>
			<td>n/a</td>
			<td>n/a&#160;</td>
			<td>Provided by your institution</td>
		</tr>
		<tr>
			<td>Small spray bottle&#160;</td>
			<td>Walmart</td>
			<td>n/a&#160;</td>
			<td>100-200 ml volume is sufficient, used for sucrose splash test</td>
		</tr>
		<tr>
			<td>Medium (mice) or large (rats) plastic tubs for swim testing</td>
			<td>Walmart</td>
			<td>variable</td>
			<td>Should be of sufficient depth that the animals cannot touch bottom (e.g., 2 feet)</td>
		</tr>
	</tbody>
</table>

an unpredictable chronic mild stress protocol for instigating depressive symptoms, behavioral changes and negative health outcomes in rodents

Cronica, lo stress non risolto è un importante fattore di rischio per lo sviluppo di depressione clinica. Mentre sono stati segnalati molti modelli preclinici di depressione indotta da stress, lo stress lieve cronico imprevedibile (UCMS) protocollo è un modello di traslazione-stabilito per indurre sintomi comportamentali comunemente associati con la depressione clinica, come anedonia, comportamento governare alterato, e impotenza appresa nei roditori. Il protocollo UCMS induce anche fisiologico (p .es., Ipercortisolemia, ipertensione) e neurologico (ad esempio, anedonia, impotenza appresa) cambia che sono clinicamente associati con la depressione. È importante sottolineare che i sintomi depressivi UCMS-indotte possono essere migliorati attraverso cronica, ma non acuta, il trattamento con SSRI comuni. Come tale, il protocollo UCMS offre molti vantaggi rispetto protocolli di stress acuto o protocolli che utilizzano fattori di stress più estremi. Il nostro protocollo prevede randomizzati, esposizioni giornaliere per 7 stre distintissors: biancheria da letto umido, rimozione di biancheria da letto, gabbia inclinazione, alterazione dei cicli di luce / buio, sollecitazioni sociali, bagno fondali bassi e predatori suoni / odori. Sottoponendo roditori 3-4 ore al giorno per questi fattori di stress lievi per 8 settimane, abbiamo dimostrato sia cambiamenti comportamentali significative e scarsi risultati per la salute per il sistema cardiovascolare. Questo approccio consente approfondito interrogatori delle alterazioni neurologiche, comportamentali e fisiologici associati alla depressione indotta da stress cronico, nonché per la sperimentazione di nuovi potenziali agenti terapeutici o strategie di intervento.

Un esempio di programma per una settimana dopo la procedura UCMS è mostrato nella tabella 1. Ogni settimana, i fattori di stress sono stati randomizzati per prevenire acclamazione e garantire imprevedibilità della sfida sforzo ogni giorno. Misure di Depressive come Comportamenti A seguito di 8 settimane di UCMS, ci sono stati cambiamenti significativi per entrambi i risultati comportamentali e fisiologici nei roditori cronicame...

Watch this Scientific Journal Video about An Unpredictable Chronic Mild Stress Protocol for Instigating Depressive Symptoms, Behavioral Changes and Negative Health Outcomes in Rodents at JoVE.com

An Unpredictable Chronic Mild Stress Protocol for Instigating Depressive Symptoms, Behavioral Changes and Negative Health Outcomes in Rodents

The unpredictable chronic mild stress (UCMS) protocol is a validated method for studying behavioral and physiological changes associated with chronic stress and depressive symptoms. Eight weeks of imposition of the UCMS protocol induces behavioral changes and poor health outcomes in rodents of either gender.

Un Imprevedibile cronica protocollo stress lieve per i sintomi depressivi Istigazione, cambiamenti comportamentali e negativi risultati di salute in Roditore

Chronic, unresolved stress is a major risk factor for the development of clinical depression. While many preclinical models of stress-induced depression ...

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22.2K Views.  West Virginia University Health Sciences Center. The unpredictable chronic mild stress (UCMS) protocol is a validated method for studying behavioral and physiological changes associated with chronic stress and depressive symptoms. Eight weeks of imposition of the UCMS protocol induces behavioral changes and poor health outcomes in rodents of either gender.

Video: An Unpredictable Chronic Mild Stress Protocol for Instigating Depressive Symptoms, Behavioral Changes and Negative Health Outcomes in Rodents

Using Chronic Social Stress to Model Postpartum Depression in Lactating Rodents

Exposure to chronic stress is a reliable predictor of depressive disorders, and social stress is a common ethologically relevant stressor in both animals and humans. However, many animal models of depression were developed in males and are not applicable or effective in studies of postpartum females. Recent studies have reported significant effects of chronic social stress during lactation, an ethologically relevant and effective stressor, on maternal behavior, growth, and behavioral neuroendocrinology. This manuscript will describe this chronic social stress paradigm using repeated exposure of a lactating dam to a novel male intruder, and the assessment of the behavioral, physiological, and neuroendocrine effects of this model. Chronic social stress (CSS) is a valuable model for studying the effects of stress on the behavior and physiology of the dam as well as her offspring and future generations. The exposure of pups to CSS can also be used as an early life stress that has long term effects on behavior, physiology, and neuroendocrinology.

This article describes the use of chronic resident intruder social stress as an ethologically relevant paradigm to model postpartum depression and anxiety in lactating rodents.

Utilizzando cronica stress sociale a modello depressione post-partum in lattazione Roditore

Exposure to chronic stress is a reliable predictor of depressive disorders, and social stress is a common ethologically relevant stressor in both animals ...

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Nest Building as an Indicator of Health and Welfare in Laboratory Mice

The minimization and alleviation of suffering has moral and scientific implications. In order to mitigate this negative experience one must be able to identify when an animal is actually in distress. Pain, illness, or distress cannot be managed if unrecognized. Evaluation of pain or illness typically involves the measurement of physiologic and behavioral indicators which are either invasive or not suitable for large scale assessment. The observation of nesting behavior shows promise as the basis of a species appropriate cage-side assessment tool for recognizing distress in mice. Here we demonstrate the utility of nest building behavior in laboratory mice as an ethologically relevant indicator of welfare. The methods presented can be successfully used to identify thermal stressors, aggressive cages, sickness, and pain. Observation of nest building behavior in mouse colonies provides a refinement to health and well-being assessment on a day to day basis.

We demonstrate the utility of nest building behavior in laboratory mice as an indicator of welfare. Nest scoring is a sensitive technique that is altered by temperature, illness, and aggression. The time to integrate into nest test (TINT) is a simple cage-side assessment that can detect postoperative pain.

The minimization and alleviation of  suffering has moral and scientific implications. In order to mitigate this  negative experience one must be able to ...

The Forced Swim Test as a Model of Depressive-like Behavior

The goal of the present protocol is to describe the forced swim test (FST), which is one of the most commonly used assays for the study of depressive-like behavior in rodents. The FST is based on the assumption that when placing an animal in a container filled with water, it will first make efforts to escape but eventually will exhibit immobility that may be considered to reflect a measure of behavioral despair. This test has been extensively used because it involves the exposure of the animals to stress, which was shown to have a role in the tendency for major depression. Additionally, the FST has been shown to share some of the factors that are influenced or altered by depression in humans, including changes in food consumption, sleep abnormalities and drug-withdrawal-induced anhedonia. The main advantages of this procedure are that it is relatively easy to perform and that its results are easily and quickly analyzed. Moreover, its sensitivity to a broad range of antidepressant drugs that makes it a suitable screening test is one of the most important features leading to its high predictive validity. Despite its appeal, this model has a number of disadvantages. First, the issue of chronic augmentation is problematic in this test because in real life patients need to be treated for at least several weeks before they experience any relief from their symptoms. Last, due to the aversiveness of the FST, it is important to take into account possible influences it might have on brain structure/function if brain analyses are to be carried out following this procedure.

This protocol describes the forced swim test, which is used for the study of depressive-like behavior in rodents. This procedure involves placing an animal in a container filled with water that eventually will lead to the exhibition of immobility behavior, which is considered to reflect behavioral despair.

Il Swim test forzato come modello di depressione-come Behavior

The goal of the present protocol is to describe the forced swim test (FST), which is one of the most commonly used assays for the study of depressive-like ...

The Double-H Maze: A Robust Behavioral Test for Learning and Memory in Rodents

Spatial cognition research in rodents typically employs the use of maze tasks, whose attributes vary from one maze to the next. These tasks vary by their behavioral flexibility and required memory duration, the number of goals and pathways, and also the overall task complexity. A confounding feature in many of these tasks is the lack of control over the strategy employed by the rodents to reach the goal, e.g., allocentric (declarative-like) or egocentric (procedural) based strategies. The double-H maze is a novel water-escape memory task that addresses this issue, by allowing the experimenter to direct the type of strategy learned during the training period. The double-H maze is a transparent device, which consists of a central alleyway with three arms protruding on both sides, along with an escape platform submerged at the extremity of one of these arms.
Rats can be trained using an allocentric strategy by alternating the start position in the maze in an unpredictable manner (see protocol 1; &#167;4.7), thus requiring them to learn the location of the platform based on the available allothetic cues. Alternatively, an egocentric learning strategy (protocol 2; &#167;4.8) can be employed by releasing the rats from the same position during each trial, until they learn the procedural pattern required to reach the goal. This task has been proven to allow for the formation of stable memory traces.
Memory can be probed following the training period in a misleading probe trial, in which the starting position for the rats alternates. Following an egocentric learning paradigm, rats typically resort to an allocentric-based strategy, but only when their initial view on the extra-maze cues differs markedly from their original position. This task is ideally suited to explore the effects of drugs/perturbations on allocentric/egocentric memory performance, as well as the interactions between these two memory systems.

The goal of this protocol is to investigate spatial cognition in rodents. The double-H water maze is a novel test, which is particularly useful to elucidate the different components of learning, consolidation and memory, as well as the interplay of memory systems.

The Double-H labirinto: una prova comportamentale robusto per l&#39;apprendimento e la memoria nei roditori

Spatial cognition research in rodents typically employs the use of maze tasks, whose attributes vary from one maze to the next. These tasks vary by their ...

A Mouse Model of Subchronic and Mild Social Defeat Stress for Understanding Stress-induced Behavioral and Physiological Deficits

Stressful life events often increase the incidence of depression in humans. To study the mechanisms of depression, the development of animal models of depression is essential. Because there are several types of depression, various animal models are needed for a deeper understanding of the disorder. Previously, a mouse model of subchronic and mild social defeat stress (sCSDS) using a modified chronic social defeat stress (CSDS) paradigm was established. In the paradigm, to reduce physical injuries from aggressors, the duration of physical contact between the aggressor and a subordinate was reduced compared to in the original CSDS paradigm. sCSDS mice showed increased body weight gain, food intake, and water intake during the stress period, and their social behaviors were suppressed after the stress period. In terms of the face validity of the stress-induced overeating and overdrinking following the increased body weight gain, the sCSDS mice may show some features related to atypical depression in humans. Thus, a mouse model of sCSDS may be useful for studying the pathogenic mechanisms underlying depression. This protocol will help establish the sCSDS mouse model, especially for studying the mechanisms underlying stress-induced weight gain and polydipsia- and hyperphagia-like symptoms.

Here, methods for developing a mouse model of subchronic and mild social defeat stress are described and used to investigate the pathogenic features of depression including hyperphagia- and polydipsia-like symptoms following increased body weight.

Un modello murino di subcronica e Mild Sconfitta sociale stress per la comprensione deficit comportamentali e fisiologiche allo stress indotto

Stressful life events often increase the incidence of depression in humans. To study the mechanisms of depression, the development of animal models of ...

Feeding Experimentation Device (FED): Construction and Validation of an Open-source Device for Measuring Food Intake in Rodents

Food intake measurements are essential for many research studies. Here, we provide a detailed description of a novel solution for measuring food intake in mice: the Feeding Experimentation Device (FED). FED is an open-source system that was designed to facilitate flexibility in food intake studies. Due to its compact and battery powered design, FED can be placed within standard home cages or other experimental equipment. Food intake measurements can also be synchronized with other equipment in real-time via FED's transistor-transistor logic (TTL) digital output, or in post-acquisition processing as FED timestamps every event with a real-time clock. When in use, a food pellet sits within FED's food well where it is monitored via an infrared beam. When the pellet is removed by the mouse, FED logs the timestamp onto its internal secure digital (SD) card and dispenses another pellet. FED can run for up to 5 days before it is necessary to charge the battery and refill the pellet hopper, minimizing human interference in data collection. Assembly of FED requires minimal engineering background, and off-the-shelf materials and electronics were prioritized in its construction. We also provide scripts for analysis of food intake and meal patterns. Finally, FED is open-source and all design and construction files are online, to facilitate modifications and improvements by other researchers.

Feeding Experimentation Device FED: Construction and Validation of an Open-source Device for Measuring Food Intake in Rodents

Feeding Experimentation Device (FED) is an open-source device for measuring food intake in mice. FED can also synchronize food intake measurements with other techniques via a real-time digital output. Here, we provide a step-by-step tutorial for the construction, validation, and usage of FED.

Alimentazione dispositivo Sperimentazione (FED): Costruzione e validazione di un dispositivo Open-source per la misurazione assunzione di cibo nei roditori

Food intake measurements are essential for many research studies. Here, we provide a detailed description of a novel solution for measuring food intake in ...

An Automated T-maze Based Apparatus and Protocol for Analyzing Delay- and Effort-based Decision Making in  Free Moving Rodents

Many neurological and psychiatric patients demonstrate difficulties and/or deficits in decision making. Rodent models are helpful to produce a deeper understanding of the neurobiological causes underlying the decision-making problems. A cost-benefit based T-maze task is used for measuring decision making in which rodents choose between a high reward arm (HRA) and a low reward arm (LRA). There are two paradigms of the T-maze decision-making task, one in which the cost is a time delay and the other in which it is physical effort. Both paradigms require a tedious and labor-intensive management of experimental animals, multiple doors, pellet reward, and arm choice recordings. In the current work, we invented an apparatus based on traditional T-maze with full automation for pellet delivery, door management and choice recordings. This automated setup can be used for the evaluation of both delay- and effort-based decision making in rodents. With the protocol described here, our lab investigated the decision-making phenotypes of multiple genetically modified mice. In the representative data, we showed that the mice with ablated medial habenular showed aversions of both delay and effort and tended to choose the immediate and effortless reward. This protocol helps to decrease the variability caused by experimenter intervention and to enhance experiment efficiency. In addition, chronic silicon probe or microelectrode recording, fiber-optic imaging and/or manipulation of neural activity can be easily applied during the decision-making task using the setup described here.

This article introduces an automated T-maze apparatus that we invented, and a protocol based on this apparatus for analyzing delay-based decision making and effort-based decision making in free moving rodents.

Un t-labirinto automatizzato basato apparato e protocollo per l'analisi basate su ritardo e sforzo decisionale nei roditori in movimento libero

Many neurological and psychiatric patients demonstrate difficulties and/or deficits in decision making. Rodent models are helpful to produce a deeper ...

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.

Il protocollo di sforzo delicato cronico imprevedibile per indurre anedonia in topi

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

Stress-Enhanced Fear Learning, a Robust Rodent Model of Post-Traumatic Stress Disorder

Fear behaviors are important for survival, but disproportionately high levels of fear can increase the vulnerability for developing psychiatric disorders such as post-traumatic stress disorder (PTSD). To understand the biological mechanisms of fear dysregulation in PTSD, it is important to start with a valid animal model of the disorder. This protocol describes the methodology required to conduct stress-enhanced fear learning (SEFL) experiments, a preclinical model of PTSD, in both rats and mice. SEFL was developed to recapitulate critical aspects of PTSD, including long-term sensitization of fear learning caused by an acute stressor. SEFL uses aspects of Pavlovian fear conditioning but produces a distinct and robust sensitized fear response far greater than normal conditional fear responses. The trauma procedure involves placing a rodent in a conditioning chamber and administering 15 unsignaled shocks randomly distributed over 90 minutes (for rat experiments; for mouse experiments, 10 unsignaled shocks randomly distributed over 60 minutes are used). On day 2, rodents are placed in a novel conditioning context where they receive a single shock; then, on day 3 they are placed back in the same context as on day 2 and tested for changes in freezing levels. Rodents that previously received the trauma display enhanced levels of freezing on the test day compared to those that received no shocks on the first day. Thus, with this model, a single highly stressful experience (the trauma) produces extreme fear of the stimuli associated with the traumatic event.

Here we describe the detailed methodology required to conduct stress-enhanced fear learning (SEFL) experiments, a preclinical model of post-traumatic stress disorder, in both rats and mice. The model utilizes aspects of Pavlovian fear conditioning and freezing as an index of enhanced fear in rodents.

Paura lo stress-Enhanced Learning, un robusto modello del roditore del disturbo da Stress post-traumatico

Fear behaviors are important for survival, but disproportionately high levels of fear can increase the vulnerability for developing psychiatric disorders ...

Novel Object Recognition and Object Location Behavioral Testing in Mice on a Budget

Ethologically relevant behavioral testing is a critical component of any study that uses mouse models to study the cognitive effects of various physiological or pathological changes. The object location task (OLT) and the novel object recognition task (NORT) are two effective behavioral tasks commonly used to reveal the function and relative health of specific brain regions involved in memory. While both of these tests exploit the inherent preference of mice for the novelty to reveal memory for previously encountered objects, the OLT primarily evaluates spatial learning, which relies heavily on hippocampal activity. The NORT, in contrast, evaluates non-spatial learning of object identity, which relies on multiple brain regions. Both tasks require an open-field-testing arena, objects with equivalent intrinsic value to mice, appropriate environmental cues, and video recording equipment and the software. Commercially available systems, while convenient, can be costly. This manuscript details a simple, cost-effective method for building the arenas and setting up the equipment necessary to perform the OLT and NORT. Furthermore, the manuscript describes an efficient testing protocol that incorporates both OLT and NORT and provides typical methods for data acquisition and analysis, as well as representative results. Successful completion of these tests can provide valuable insight into the memory function of various mouse model systems and appraise the underlying neural regions that support these functions.

Here we provide a protocol which includes comprehensive instructions for the economical establishment of murine object location and novel object recognition behavioral testing, including the design, cost, and construction of required equipment as well as execution of behavioral testing, data collection, and analysis.