Gli autori ringraziano Drs. Kentaro Nagaoka (Tokyo University of Agriculture and Technology) e Wataru Iio (Prefettura di Ibaraki) per la discussione utile. Questa ricerca è stata sostenuta in parte da Ibaraki cooperazione universitaria tra agricoltura e Scienze Mediche (IUCAM) (Il MEXT, Giappone) e il progetto di ricerca sullo sviluppo dei prodotti agricoli e di alimenti con benefici che promuovono la salute (NARO) (Il MAFF, Giappone) .

Gli studi su animali sono stati approvati dal e il rispetto delle linee guida sia della cura degli animali e del Comitato uso di Ibaraki University e il Ministero della Cultura Istruzione, Sport, Scienza, Tecnologia e (MEXT), Giappone (Notifica No.71). Una panoramica completa del protocollo è mostrato in Figura 1. 1. Apparato <ol><li> Preparare due tipi di gabbie: una gabbia singola (larghezza [W] × profondità [D] × altezza [H] = 143 millimetri x 293 millimetri x 148 mm), e una &quot;sconfitta sociale (SD)&quot; gabbia (L × P × H = 220 mm × 320 mm × 135 millimetri). </li><li> Come mostrato in figura 2, dividere la gabbia SD in due scomparti con un acrilico trasparente bordo divisore (5 mm di spessore) con 15 fori circolari (3 × 5 matrice: 8 mm di diametro). </li><li> Ottenere i chip di legno-rasatura a base di abeti, pellet cibo purificato-dieta, e una bottiglia di acqua potabile. Inoltre, ottenere asciugamani di carta, una maschera e guanti in lattice. </li&gt; <li> Per la prova interazione sociale, preparare un&#39;arena campo aperto (L × P × = 40 centimetri × 40 cm x 40 cm) in grigio polivinilcloruro, un peso in acciaio (190 g), e una scatola di interazione di plastica (W × P × = 10 × 10 cm × 13 cm, 100 g) con tre finestre di rete metallica (L x P = 5 centimetri x 5 cm) (Figura 3). </li><li> Posizionare una telecamera CCD (2,8-12 mm; F ​​= 1: 1,4; 1/3 di pollice CCD) e un sistema automatizzato di monitoraggio nella sala prove del comportamento nella struttura animali. Inserire un ripiano appropriato per le gabbie di topo nella camera test comportamentali abituare i topi all&#39;ambiente della camera di prova per almeno 30 min. </li></ol> 2. assuefazione all&#39;ambiente <ol><li> Utilizzare maschio C57BL / 6JJmsSlc (B6) topi che sono 7 settimane di vita e di sesso maschile Slc: ICR (ICR) i topi, che sono più di 5 mesi e consegnare alla struttura da una società di allevamento di animali. </li><li> Indipendentemente spostare due gruppi di topi B6 (n= 12 in ciascun gruppo) per l&#39;impianto; utilizzare un gruppo (screener B6 topi) per lo screening di aggressivi topi ICR e un altro gruppo (soggetto B6 topi) per lo sviluppo del modello sCSDS. </li><li> Introdurre topi ICR (n = 12) per l&#39;impianto, al fine di selezionare i loro comportamenti aggressivi. </li><li> Casa i topi individualmente in gabbie singole per 1 settimana sotto un ciclo luce-buio di 12 ore (circa luce fluorescente 100 lux, si accende alle 08:00) con temperatura costante (intorno a 23 ° C) e umidità (circa 40%) al fine di abituare all&#39;ambiente. Partizionare ogni gabbia posizionando tavole di plastica di colore bianco tra le gabbie in modo che i topi non sono influenzati dai comportamenti dei topi vicini. </li><li> Fai pellet cibo purificato-dieta e acqua ad osmosi inversa disponibili ad libitum. Utilizzare AIN-93G chow perché gli ingredienti di altri pellet dieta non purificate possono variare. </li><li> Misurare il peso corporeo, l&#39;assunzione di cibo (FI) e di acqua (WI) dei topi ogni giorno. Calcolare il baumento di peso ody (BWG) dal giorno iniziale. </li></ol> 3. Proiezione di aggressivo ICR Mice <ol><li> Dopo assuefazione per 1 settimana come già detto, schermare i topi ICR (residenti) utilizzando test di soggiornante di intrusione per una prova di 3 minuti con i topi vagliatore B6 (antintrusione, 8 settimane di età) nel pomeriggio (14: 00-17: 00) sotto illuminazione di circa 300 lux nella stanza custodia. <ol><li> In particolare, verificare ogni mouse ICR per tre prove al giorno per 5 giorni consecutivi (15 prove in totale) verso molteplici topi B6 possibile per trovare quale i topi ICR mostrano elevata aggressività verso gli intrusi. Durante le prove, registrare la latenza attacco e la durata del comportamento aggressivo (movimenti rapidi con attacco mordere). </li></ol></li><li> Identificare iper-aggressivo ICR topi controllando le ferite danno sui topi intruso B6 dopo ogni prova. </li><li> Calcolare il rapporto di prove in cui la latenza attacco è inferiore a 30 sec come primo indice del punteggio aggressione. </li><li> Calculate il rapporto di prove in cui la latenza attacco è meno di 3 minuti in un secondo indice del punteggio aggressione. </li><li> Valutare i punteggi aggressione del primo indice. Utilizzare il secondo indice quando il primo indice è uguale. </li><li> Selezionare i topi ICR che avevano punteggi più alti aggressione senza comportamenti iper-aggressivo come aggressivi topi ICR. Utilizzare i topi ICR aggressivi ripetutamente per tutta la prossima serie di esperimenti fino a circa 12 mesi di età; tuttavia, eseguire il processo di screening per aggressivi topi ICR come descritto in precedenza prima di ogni esperimento per confermare l&#39;aggressività dei topi ICR. </li><li> Dopo ogni sessione dello screening, registrare il grado di lesione dei topi screener B6. Se il mouse è ferito, isolare il mouse in una singola gabbia e osservare i progressi controllando il suo guadagno di peso corporeo, l&#39;assunzione di cibo, e l&#39;assunzione di acqua. In caso di gravi ferite che influenzano la fisiologia e comportamento nei topi, eutanasia li secondo I localiLinee guida ACUC. </li></ol> 4. sCSDS <ol><li> Introdurre soggetto topi B6 7 giorni prima (giorno -6) il giorno di esposizione iniziale di stress (giorno 1) e dalla casa dei topi singolarmente in gabbie singole loro abituare all&#39;ambiente. </li><li> Tre giorni prima del giorno 1 (il giorno -2), spostare i topi ICR aggressive in un compartimento di ogni gabbia SD, che è diviso da un divisore acrilica per consentire ai topi stabiliscono loro territori nelle gabbie SD (lo stesso numero di soggetti B6 topi). </li><li> Dividere le gabbie SD utilizzando schede partizionamento di colore bianco, come nella condizione singola gabbia sopra descritto. </li><li> Per i topi B6 non stressati controllo, preparare gabbie SD (metà del numero di topi) per mantenere i topi a coppie; posizionare due topi in ciascun vano diviso per il divisore in una gabbia SD per 10 giorni. </li><li> Dopo le misurazioni quotidiane BWG, FI, e WI il giorno 1, topi posto soggetto B6 in uno degli scomparti delle gabbie SD (casa-gabbia di residenti) nel afternoon (14: 00-16: 00) con illuminazione di circa 300 lux nella stanza alloggiamento e misurare il tempo di contatto fisico dal primo morso attacco per completare il test residente-intruso. </li><li> Il giorno 1, impostare il tempo di contatto fisico a 5 min dal primo morso attacco e hanno osservatori contare il numero di punture di attacco da topi ICR che sono dirette preferenzialmente sul retro o fianchi dell&#39;avversario come descritto da Miczek et al. 20 determinare il grado di stress fisico a topi B6. </li><li> Dopo il tempo di contatto fisico, salvataggio soggetti topi B6, e controllare e registrare il loro stato di pelliccia e ferite. Quindi, posizionare i topi B6 in un altro scompartimento accanto ai topi ICR nelle gabbie SD fino a esposizione a stress fisico il giorno successivo. </li><li> Poiché il divisore in acrilico nella gabbia SD è trasparente e contiene buchi, esporre soggetti topi B6 a vari stress emotivi, tra cui visivi, uditivi, olfattivi e stimoli, dalle topi ICR nel vano vicinazione della gabbia SD per 24 ore ogni giorno. </li><li> Misurare la BWG, FI e WI di topi B6 controllo quotidiano e poi metterli in ciascun comparto nelle gabbie SD per 1 giorno. </li><li> Il giorno 2, dopo le misurazioni giornaliere del BWG, FI, e WI, introdurre soggetti topi B6 nei territori di altri topi ICR per esporli a stress fisico. </li><li> Impostare la durata del contatto fisico a 4,5 minuti dal primo boccone attacco il giorno 2, e contare il numero di punture di attacco. </li><li> Spostare i topi B6 di controllo in diversi comparti e modificare le combinazioni di coppie allo scopo di mischiare il posizionamento e partner di coppie ogni giorno. </li><li> Diminuire la durata del contatto fisico di 0,5 minuti al giorno, in modo che la durata del giorno 10 è impostato a 0,5 min dal primo morso attacco. </li><li> Il giorno 7, sostituire circa la metà dei trucioli in tutti i compartimenti nelle gabbie SD. </li><li> Dopo l&#39;esposizione all&#39;ultima stress fisico al giorno 10, spostare soggette topi B6 in gabbie singole. Simile ai topi B6 soggetto, spostare i topi di controllo B6 in gabbie singole il giorno 10. </li><li> Se un mouse ICR non mostra eventuali punture di attacco fino a 5 minuti in ogni prova nei giorni da 1 a 10, terminare il processo. Sostituire il mouse con i pezzi di topi ICR ICR aggressivi e condurre un processo alternativa per il mouse soggetto B6. </li><li> Se il mouse è ferito, isolare il mouse in una singola gabbia e osservare i progressi controllando il suo guadagno di peso corporeo, l&#39;assunzione di cibo, e l&#39;assunzione di acqua. Segui le tue linee guida IACUC per l&#39;analgesia. In caso di gravi ferite che influenzano la fisiologia e comportamento nei topi, eutanasia loro secondo le direttive IACUC locali. </li></ol> 5. Evitare di prova sociale (interazione sociale Test) <ol><li> Effettuare test comportamentali per evitamento sociale di mattina (09: 00-12: 00) il giorno 11. </li><li> Trenta minuti prima del test, trasferire le gabbie di controllo e soggetto B6 topi su un ripiano (penombra inferiore a 1 lux) nella comportamentalesala prove sotto illuminazione inferiore a 20 lux per permettere loro di abituare all&#39;ambiente. </li><li> Per ridurre ordine effetti, alternativamente test di controllo e soggetto B6 topi. </li><li> Pulire un&#39;arena campo aperto (illuminato con una luce di 20 lux al centro del campo) e una scatola di interazione plastica con carta assorbente imbevuti con etanolo al 70% prima della prova comportamentale per ogni mouse per rimuovere le feci, urine, e gli odori . </li><li> Inserire un topo ICR sconosciuta (non utilizzato come un aggressore) da una singola gabbia vicino all&#39;apparecchiatura campo aperto. </li><li> Posizionare il mouse B6 nel campo aperto della scatola interazione vacante, come illustrato nella figura 4. Monitorare e analizzare il comportamento di 2.5 min utilizzando il sistema automatico di analisi (descritto al punto 5.10). </li><li> Dopo la prima prova, togliere il mouse B6 dal campo e metterlo nella sua gabbia casa. </li><li> A seguito di questo, posizionare il mouse ICR non familiare maschio (target sociale) nella casella di interazione e quindi introdurre il B6il mouse per i campi e controllare il suo comportamento per 2.5 min. </li><li> Dopo il secondo processo, restituire sia la B6 e topi ICR alle loro gabbie a casa, e pulire il campo e la scatola di interazione di cui sopra. </li><li> Ripetere questi passaggi per ogni mouse B6 per testare il comportamento sociale. Nel corso di ogni prova, registrare filmati top-vista con una camera CCD. <ol><li> Dopo le prove comportamentali, calcolare il tempo trascorso nella zona di interazione (in secondi) e nella zona d&#39;angolo (in secondi) per ciascuna prova come mostrato in Goto et al. 13. Giudicare la posizione del mouse in base al centro del mouse. </li><li> Calcolare il punteggio di interazione sociale come 100 × (tempo di interazione, con target) / (tempo di interazione, senza bersaglio), seguendo i metodi pubblicati in Krishnan et al. 21. </li></ol></li></ol>

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Renal Physiol. 290 (2), F478-F485 (2006).</li><li>Flores, P., 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=Schedule-Induced+Polydipsia:+Searching+for+the+Endophenotype+of+Compulsive+Behavior.">Schedule-Induced Polydipsia: Searching for the Endophenotype of Compulsive Behavior.</a> World J. Neurosci. 4, 253-260 (2014).</li><li>Cryan, J. F., Dinan, T. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mind-altering+microorganisms:+the+impact+of+the+gut+microbiota+on+brain+and+and+behaviour.">Mind-altering microorganisms: the impact of the gut microbiota on brain and and behaviour.</a> Nat. Rev. Neurosci. 13 (10), 701-712 (2012).</li></ol>

Gli autori dichiarano di non avere interessi finanziari in competizione.

Ci sono state differenze definitive del peso corporeo tra sCSDS topi e topi CSD sottoposti al protocollo CSDS standard (5-10 min di contatto fisico con gli aggressori al giorno) 19. I sCSDS topi hanno mostrato un aumento BWG durante il periodo di stress, mentre i topi normali CSD hanno mostrato una diminuzione del peso corporeo durante il periodo di stress 21,22,23. Vi è una differenza sostanziale tra i due protocolli in termini di durata totale di contatto fisico con aggressori durante il periodo...

Molti tipi di eventi stressanti si verificano durante tutto la vita di esseri umani. Lo stress eccessivo porta spesso a conseguenze fisiologiche nocivi nell&#39;uomo e negli animali. Negli esseri umani, gli eventi stressanti sono i principali fattori di rischio per la precipitazione disturbi psichiatrici come la depressione 1. Un Global Burden of Disease studio ha indicato che la depressione è uno dei disturbi più invalidanti, in termini di anni di vita adattati alla disabilità (DALY) e gli anni vissuti con disabilità 2. Inoltre, la depressione rappresenta la maggior parte del DALY suicidi 3. Le persone che soffrono di depressione hanno difficoltà a gestire la propria vita, e di conseguenza, la qualità della vita peggiora spesso. Pertanto, vi è una forte necessità di sviluppare terapie efficaci per migliorare la qualità della vita in questi pazienti. Molti studi sono stati condotti sui principali disturbi depressivi, e hanno rivelato che il contributo genetico alla malattia susceptibility è circa il 30-40%, che si spiega con gli effetti di loci multipli di piccoli effetti 4. A causa dei complessi meccanismi patogenetici alla base della depressione, l&#39;eziologia dettagliata della malattia rimane inafferrabile. Rapporti clinici indicano che ci sono sottotipi di depressione, come malinconica e atipico depressione 5, da cui risultano ridotti e aumento del peso corporeo, rispettivamente, 6. Anche se il 25-30% e il 15-30% dei pazienti con depressione rappresentano caratteristiche melanconici e atipici puri, rispettivamente, la maggior parte di loro hanno caratteristiche miste di entrambi i sottotipi 7. Pertanto, la depressione ha una vasta gamma di sintomi. Al fine di trovare biomarker e sviluppare terapie oggettivi per i vari tipi di depressione negli esseri umani, è importante stabilire diversi modelli animali differenti di depressione 8. Modelli animali di depressione sono stati stabiliti utilizzando diversi approcci compresi imparatoimpotenza, lo stress cronico lieve imprevedibile, e lo stress sconfitta sociale cronico (CSDS) 9-12. Toyoda e colleghi stabiliti i modelli CSD di ratti e topi 13-17, al fine di chiarire il metabolismo e comportamenti che sono associati con la depressione. Dato che i modelli animali di depressione sono valutati da validità faccia 18, il contesto in cui è stabilito il modello è importante. Inoltre, Golden et al. 19 hanno riportato i metodi per la creazione di topi CSD in dettaglio. E &#39;noto che i deficit nel comportamento sociale dei topi CSD possono essere recuperati da un trattamento cronico, ma non per il trattamento acuto, con antidepressivi, e che condividono sintomi simili a quelli in pazienti con depressione in termini di regolamentazione di brain-derived neurotrophic fattore 6. Goto et al. 13 precedentemente sviluppato la subcronica e mite lo stress sconfitta sociale (sCSDS) modello di topo, modificando i metodidi Golden et al. 19. I sCSDS topi mostrano sintomi polydipsia- e iperfagia come seguenti aumenti di peso corporeo e una maggiore corpo contenuto di acqua 13. In questa relazione, il protocollo per stabilire il modello del mouse sCSDS è fornito e discutiamo l&#39;utilità di questo modello.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>single cage</td>
			<td>Charles River Laboratories Japan</td>
			<td></td>
			<td>width [W] &#215; depth [D] &#215; height [H] = 143 &#215; 293 &#215; 148 mm</td>
		</tr>
		<tr>
			<td>M cage</td>
			<td>Natsume Seisakusho</td>
			<td></td>
			<td>W &#215; D &#215; H = 220 &#215; 320 &#215; 135 mm</td>
		</tr>
		<tr>
			<td>Whiteflake</td>
			<td>Charles River Laboratories Japan</td>
			<td></td>
			<td>Wood-shaving chips made from spruce trees</td>
		</tr>
		<tr>
			<td>AIN-93G</td>
			<td>Oriental Yeast</td>
			<td></td>
			<td>purified-diet food pellets</td>
		</tr>
		<tr>
			<td>Kimtowel</td>
			<td>Nippon Paper Crecia Co.</td>
			<td></td>
			<td>Paper towels</td>
		</tr>
		<tr>
			<td>open-field arena</td>
			<td>O&#8217;Hara &#38; Co.</td>
			<td></td>
			<td>made of gray polyvinylchloride</td>
		</tr>
	</tbody>
</table>

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.

Per monitorare il grado di stress fisico per nei periodi di 10 giorni, il numero di punture di attacco di topi ICR stato contato manualmente da un ricercatore. Figura 5A indica i singoli valori per il numero di punture attacco ricevuti. C&#39;era una notevole variabilità nella fase iniziale (circa 10-120 punture al giorno 1), ma questa variabilità è stata ridotta nella fase successiva (circa 5-20 morsi sul giorno 10). La figura 5B indica che il numero medio di punture di attacco rice...

Watch this Scientific Journal Video about A Mouse Model of Subchronic and Mild Social Defeat Stress for Understanding Stress-induced Behavioral and Physiological Deficits at JoVE.com

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

Qui, metodi per sviluppare un modello di topo di subcronica e mite lo stress sconfitta sociale sono descritti e usati per studiare le caratteristiche patogene di depressione, tra cui hyperphagia- e sintomi simil-polidipsia seguenti aumento di peso corporeo.

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 ...

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JoVE Journal

Behavior

19.4K Views.  Ibaraki University.  Qui, metodi per sviluppare un modello di topo di subcronica e mite lo stress sconfitta sociale sono descritti e usati per studiare le caratteristiche patogene di depressione, tra cui hyperphagia- e sintomi simil-polidipsia seguenti aumento di peso corporeo. 

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

The Resident-intruder Paradigm: A Standardized Test for Aggression, Violence and Social Stress

This video publication explains in detail the experimental protocol of the resident-intruder paradigm in rats. This test is a standardized method to measure offensive aggression and defensive behavior in a semi natural setting. The most important behavioral elements performed by the resident and the intruder are demonstrated in the video and illustrated using artistic drawings. The use of the resident intruder paradigm for acute and chronic social stress experiments is explained as well. Finally, some brief tests and criteria are presented to distinguish aggression from its more violent and pathological forms.

This video shows the resident-intruder paradigm in rats. This test is a standardized method to measure offensive aggression, defensive behavior and violence in a semi-natural setting. The use of the paradigm for social stress experiments is explained as well.

Il Paradigma residente-intruso: un test standardizzato per aggressione, di violenza e di stress sociale

This video publication explains in detail the experimental protocol of the resident-intruder paradigm in rats. This test is a standardized method to ...

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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 ...

Moderate Prenatal Alcohol Exposure and Quantification of Social Behavior in Adult Rats

Alterations in social behavior are among the major negative consequences observed in children with Fetal Alcohol Spectrum Disorders (FASDs). Several independent laboratories have demonstrated robust alterations in the social behavior of rodents exposed to alcohol during brain development across a wide range of exposure durations, timing, doses, and ages at the time of behavioral quantification. Prior work from this laboratory has identified reliable alterations in specific forms of social interaction following moderate prenatal alcohol exposure (PAE) in the rat that persist well into adulthood, including increased wrestling and decreased investigation. These behavioral alterations have been useful in identifying neural circuits altered by moderate PAE1, and may hold importance for progressing toward a more complete understanding of the neural bases of PAE-related alterations in social behavior. This paper describes procedures for performing moderate PAE in which rat dams voluntarily consume ethanol or saccharin (control) throughout gestation, and measurement of social behaviors in adult offspring.

The goal of the protocol presented here is to describe procedures to expose rats to moderate levels of alcohol during prenatal brain development and to quantify resulting alterations in social behavior during adulthood.

Moderata esposizione prenatale di alcool e quantificazione di comportamento sociale in ratti adulti

Alterations in social behavior are among the major negative consequences observed in children with Fetal Alcohol Spectrum Disorders (FASDs). Several ...

The Social Dimension of Stress: Experimental Manipulations of Social Support and Social Identity in the Trier Social Stress Test

In many situations humans are influenced by the behavior of other people and their relationships with them. For example, in stressful situations supportive behavior of other people as well as positive social relationships can act as powerful resources to cope with stress. In order to study the interplay between these variables, this protocol describes two effective experimental manipulations of social relationships and supportive behavior in the laboratory. In the present article, these two manipulations are implemented in the Trier Social Stress Test (TSST)&#8212;a standard stress induction paradigm in which participants are subjected to a simulated job interview. More precisely, we propose (a) a manipulation of the relationship between different protagonists in the TSST by making a shared social identity salient and (b) a manipulation of the behavior of the TSST-selection committee, which acts either supportively or unsupportively. These two experimental manipulations are designed in a modular fashion and can be applied independently of each other but can also be combined. Moreover, these two manipulations can also be integrated into other stress protocols and into other standardized social interactions such as trust games, negotiation tasks, or other group tasks.

Previous research on the social dimension of stress has focused on two important variables: social identity and social support. This protocol introduces an effective experimental manipulation of these two social variables and describes their implementation in a standard stress induction paradigm (Trier Social Stress Test).

La dimensione sociale dello stress: manipolazioni sperimentali di sostegno sociale e identità sociale nella Trier Social Stress Test

In many situations humans are influenced by the behavior of other people and their relationships with them. For example, in stressful situations ...

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

Chronic, unresolved stress is a major risk factor for the development of clinical depression. While many preclinical models of stress-induced depression have been reported, the unpredictable chronic mild stress (UCMS) protocol is an established translationally-relevant model for inducing behavioral symptoms commonly associated with clinical depression, such as anhedonia, altered grooming behavior, and learned helplessness in rodents. The UCMS protocol also induces physiological (e.g., hypercortisolemia, hypertension) and neurological (e.g., anhedonia, learned helplessness) changes that are clinically associated with depression. Importantly, UCMS-induced depressive symptoms can be ameliorated through chronic, but not acute, treatment with common SSRIs. As such, the UCMS protocol offers many advantages over acute stress protocols or protocols that utilize more extreme stressors. Our protocol involves randomized, daily exposures to 7 distinct stressors: damp bedding, removal of bedding, cage tilt, alteration of light/dark cycles, social stresses, shallow water bath, and predator sounds/smells. By subjecting rodents 3-4 hr daily to these mild stressors for 8 weeks, we demonstrate both significant behavioral changes and poor health outcomes to the cardiovascular system. This approach allows for in-depth interrogation of the neurological, behavioral, and physiological alterations associated with chronic stress-induced depression, as well as for testing of new potential therapeutic agents or intervention strategies.

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 ...

Detecting Behavioral Deficits in Rats After Traumatic Brain Injury

With the increasing incidence of traumatic brain injury (TBI) in both civilian and military populations, TBI is now considered a chronic disease; however, few studies have investigated the long-term effects of injury in rodent models of TBI. Shown here are behavioral measures that are well-established in TBI research for times early after injury, such as two weeks, until two months. Some of these methods have previously been used at later times after injury, up to one year, but by very few laboratories. The methods demonstrated here are a short neurological assessment to test reflexes, a Beam-Balance to test balance, a Beam-Walk to test balance and motor coordination, and a working memory version of the Morris water maze that can be sensitive to deficits in reference memory. Male rats were handled and pre-trained to neurological, balance, and motor coordination tests prior to receiving parasagittal fluid percussion injury (FPI) or sham injury. Rats can be tested on the short neurological assessment (neuroscore), the beam-balance, and the Beam-Walk multiple times, while testing on the water maze can only be done once. This difference is because rats can remember the task, thus confounding the results if repeated testing is attempted in the same animal. When testing from one to three days after injury, significant differences are detected in all three non-cognitive tasks. However, differences in the Beam-Walk task were not detectable at later time points (after 3 months). Deficits were detected at 3 months in the Beam-Balance and at 6 months in the neuroscore. Deficits in working memory were detected out to 12 months after injury, and a deficit in a reference memory first appeared at 12 months. Thus, standard behavioral tests can be useful measures of persistent behavioral deficits after FPI.

The goal of the behavioral tests presented here is to detect functional deficits in rats after traumatic brain injury. Four specific tests are presented that detect deficits in behaviors to reflect the damage to specific brain areas at times extending to one year after injury.

Rilevare i deficit comportamentistici in ratti dopo la ferita di cervello traumatica

With the increasing incidence of traumatic brain injury (TBI) in both civilian and military populations, TBI is now considered a chronic disease; however, ...

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 ...

A Visual Guide for Studying Behavioral Defenses to Pathogen Attacks in Leaf-Cutting Ants

The complex lifestyle, evolutionary history of advanced cooperation, and disease defenses of leaf-cutting ants are well studied. Although numerous studies have described the behaviors connected with disease defense, and the associated use of chemicals and antimicrobials, no common visual reference has been made. The main aim of this study was to record short clips of behaviors involved in disease defense, both prophylactically and directly targeted towards an antagonist of the colony following infection. To do so we used an infection experiment, with sub-colonies of the leaf-cutting ant species Acromyrmex echinatior, and the most significant known pathogenic threat to the ants&#39; fungal crop (Leucoagaricus gongylophorus), a specialized pathogenic fungus in the genus Escovopsis. We filmed and compared infected and uninfected colonies, at both early and more advanced stages of infection. We quantified key defensive behaviors across treatments and show that the behavioral response to pathogen attack likely varies between different castes of worker ants, and between early and late detection of a threat. Based on these recordings we have made a library of behavioral clips, accompanied by definitions of the main individual defensive behaviors. We anticipate that such a guide can provide a common frame of reference for other researchers working in this field, to recognize and study these behaviors, and also provide greater scope for comparing different studies to ultimately help better understand the role these behaviors play in disease defense.

We present a visual guide to disease defense behaviors in leaf-cutting ants, with individual clips and accompanying definitions, illustrated in an experimental infection scenario. Our main aim is to help other researchers recognize key defensive behaviors and to provide a common understanding for future research in this field.

Una guida visiva per studiare le difese del comportamento patogeno attacca in formiche tagliafoglie

The complex lifestyle, evolutionary history of advanced cooperation, and disease defenses of leaf-cutting ants are well studied. Although numerous studies ...

Nest Building Behavior as an Early Indicator of Behavioral Deficits in Mice

Nest building is an innate behavior in male and female rodents, even when raised in laboratory settings. As such, many researchers provide rodents synthetic and/or natural materials (such as twine, tissue, cotton, paper, and hay) as a gauge of their overall well-being and as an ancillary assessment to predict the possible decline in cognition. Typically, changes in nesting behaviors, such as failure to create a nest, indicate a change in health or welfare. In addition, nesting behavior is sensitive to many environmental and physiological challenges, as well as many genetic mutations underlying pathological disease states. The following protocol describes a nesting behavior paradigm that explores the usage of four types of nesting material. In addition, the protocol utilizes intraclass correlations to demonstrate that inter-rater reliability is higher when nests are constructed out of shredded paper compared to other common nesting materials such as cotton squares, paper twists, and soft cob bedding. The chosen methodology and statistical considerations (i.e., intraclass correlation) for this assay may be of interest for those conducting experiments assessing the quality of living of mice.&#160;

Here, we present a protocol to quantify nest building behavior in mice, which is known to be impaired in several neurological disorders and diseases. This protocol examines the utility of four materials and offers the opportunity to quantify the rater agreement in scoring, improving the validity and reliability of the assay.