Support provided by grant AA019462 to DAH and AA019884 to DDS.

Tutte le procedure qui e nel video allegato descritti sono stati approvati dalla cura e utilizzo dei Comitati animali istituzionale del Health Sciences Center e il campus principale della University of New Mexico. 1. prenatale Etanolo Esposizione <ol><li> Ottenere tutti i materiali e le sostanze chimiche necessarie: Saccarina idrato sale di sodio, 190 etanolo prova (alcool al 95% in volume), tubi di bere vetro perline, gomma naturale bianco # 4 tappo un foro, 1 &quot;punto di curvatura tubi-ball, governanti carta stampata da www .vendian.org / mncharity / dir3 / paper_rulers /. </li><li> Ottenere provati ratti allevatore adulto da un fornitore o fonte alternativa. Utilizzare allevatori lungo Evans per i metodi ei dati rappresentativi presentati in questo protocollo. Le femmine pesano 125-150 g e 6-7 settimane di età all&#39;arrivo tale che essi sono di circa 9-10 settimane al momento della riproduzione (passo 1.5). Assicurarsi che tutti i futuri dighe sono primipare. Assicurarsi che i maschi sono 12settimane di età in arrivo e 15 settimane all&#39;inizio del protocollo allevamento. </li><li> Casa gli animali singolarmente in gabbie di plastica a 22 ° C su un rovescio 12 ore di luce / buio calendario (si accende 21:00-9:00 hr). Fornire l&#39;accesso a cibo e acqua del rubinetto ad libitum per tutto lo studio, anche durante tutte le sessioni di bere. Lasciare almeno 1 settimana per acclimatazione alla struttura prima di procedere alla fase 1.4. </li><li> Ottenere peso corporeo di base per ogni ratto femmina. </li><li> Valutare i livelli di consumo pre-gravidanza nei ratti femmina per 2 settimane. Seguire i passaggi 1.5.1-1.5.6 in ordine. <ol><li> Nei giorni 1 e 2 riempire un tubo potabile contenente 0,066% soluzione saccarina (senza etanolo) in acqua di rubinetto. Per quantificare il consumo di apporre un righello di carta con precisione mm al tubo di bere prima di riempirlo. Utilizzare questo metodo in quanto riduce gli errori in misura associata pesatura tubi per quantificare il consumo di etanolo. Riempire il tubo per i 20 mm del righello. Inizia quattro ore drinkisessioni ng (passi 1.5.1-1.5.6 e 1.7) 1 ora dopo l&#39;inizio della fase di buio (10:00 ore), quando i livelli di attività e bere sono più elevati. </li><li> Al termine di ogni sessione potabile quantificare il volume della soluzione di saccarina che è stato consumato. Per facilitare la misurazione, determinare il volume di soluzione per mm sul righello in anticipo e convertire mm volumi. Per i tubi raccomandate qui ogni mm corrisponde a 0,366 ml. </li><li> Nei giorni 3 e 4 riempire un tubo con 2,5% di etanolo (v / v) e la soluzione di saccarina 0,066% in acqua di rubinetto. Al termine di ogni sessione di quantificare la quantità di soluzione di etanolo (peso per kg di peso corporeo) che è stato consumato. </li><li> Il giorno 5 e successivamente riempire un tubo con 5% di etanolo (v / v) e la soluzione di saccarina 0,066% in acqua di rubinetto. Al termine di ogni sessione di quantificare la quantità di soluzione di etanolo (peso per kg di peso corporeo) che è stato consumato. </li><li> Al completamento della fase di pre-gravidanza potabile pesare i ratti e calcolare il consum etanolo medioption e deviazione standard per l&#39;intero gruppo. Rimuovere i ratti per il che significa il consumo è maggiore di 1 deviazione standard sopra o sotto il gruppo media dallo studio per ridurre la variabilità nel bere volontaria durante la gravidanza. </li><li> Assegnare i rimanenti ratti sia al controllo saccarina o condizioni PAE tali che i livelli bere pre-gravidanza sono abbinati il ​​più possibile per i due gruppi. </li></ol></li><li> Entro 1-14 giorni coppia ogni ratto femmina con un allevatore di sesso maschile provata. Ratti femmina non consumano etanolo durante la fase di riproduzione. Confermare la gravidanza dalla presenza di un tappo vaginale, pesare il ratto femmina, e la casa dei suoi individualmente. Questo è definito come il giorno di gestazione 1. Facoltativamente, lasciare i ratti femmina con un allevatore di sesso maschile per un massimo di 5 giorni, dopo che rimuovono le femmine dallo studio. NOTA: poiché il consumo di etanolo durante la gravidanza inizia alle 10:00 ora il gestazionale giorno 1 di ispezione per la presenza di un tappo vaginale deve essere eseguitaprima di questo tempo. </li><li> Fornire soluzioni etanolo o saccarina per 4 ore al giorno (1.000 a 1.400 ore) per la durata della gravidanza. <ol><li> A partire dal giorno di gestazione 1 forniscono il topo femmina con uno 0% o 5% di etanolo e la soluzione di saccarina 0,066% in acqua di rubinetto in base alla assegnazione di gruppo. Assicurarsi che il volume di soluzione di etanolo 0% fornito ai ratti di controllo saccarina corrisponde al volume medio di soluzione di etanolo al 5% consumata dai topi etanolo. Al termine di ogni sessione di quantificare la quantità di soluzione di etanolo (peso per kg di peso corporeo) o la saccarina che è stato consumato. </li><li> Pesare dighe ratto settimanale per valutare l&#39;aumento di peso materno. </li><li> Fornire cibo e acqua del rubinetto in ogni momento, tra cui le bevute. </li><li> Cessate le procedure di esposizione etanolo quando la prole sono nati. Registrare il numero di cuccioli vivi e pesi dei cuccioli alla nascita. Designare il giorno della nascita come postnatale giorno 0. </li><li> Abbattere la cucciolata di 10 cuccioli in giro giorno postnatale 2-3. Tentativo to mantenere una parità di rapporto maschi femmine in ogni cucciolata. Se questo non è possibile, abbattere a 10 cuccioli con un numero disuguale di maschi e femmine. </li><li> Registra pesi cuccioli 2-3 giorni dopo la nascita. </li><li> Svezzare gli animali al giorno circa 21-24 postnatale e la casa a coppie dello stesso sesso con un animale della stessa condizione trattamento prenatale. Non utilizzare più di 1-2 ratti da ogni disordine per esperimento per limitare i potenziali effetti del disordine. </li></ol></li></ol> 2. Social Behavior <ol><li> Ottenere e preparare tutti i materiali e le attrezzature necessarie. <ol><li> Ottenere un apparato per l&#39;interazione sociale. Usare una camera con una parte superiore aperta costruito di materiale facile da pulire e sterilizzare. La parte anteriore dell&#39;apparecchio deve essere coperto con plastica rigida trasparente (~ spessore 2 mm) per le riprese. Linea le pareti interne e il pavimento con plastica trasparente (~ 2 mm di spessore) per aiutare nella pulizia e controllo degli odori. Specchi svolge lungo il wa interno posteriorell per aiutare l&#39;analisi. NOTA: Per i dati rappresentativi riportati qui una camera personalizzato (95 cm di lunghezza × 47 centimetri di larghezza di altezza × 43 cm), con un top aperto e trasparente di plastica rigida (2 mm di spessore) che copre i fianchi interni e piano dell&#39;apparato è stato utilizzato. Specchi sono stati collocati lungo la parete di fondo. Le dimensioni e materiali specifici non sono critici per la misurazione del comportamento sociale, tuttavia, è consigliabile che le dimensioni essere sufficientemente grande da garantire che autentica interazione sociale può essere distinto da altri comportamenti. Cioè, l&#39;apparato deve essere abbastanza grande in modo che gli animali non sono sempre in stretta vicinanza l&#39;uno all&#39;altro. </li><li> Ottenere telecamere in grado di registrare in condizioni di luce bassa o come non tutte le riprese è condotta con illuminazione poco o nessun ambiente all&#39;interno dello spettro visibile. Assicurarsi che la fotocamera ha una risoluzione elevata nello spettro infrarosso, tuttavia, qualsiasi fotocamera in grado di registrare in condizioni di oscurità nella sua nativamodalità notte o con l&#39;aggiunta di illuminatori a infrarossi dovrebbe essere sufficiente. </li><li> Posizione illuminatori infrarossi intorno all&#39;apparecchio per migliorare l&#39;illuminazione dell&#39;apparecchio nella registrazione video. </li><li> Ottenere cippato grado di laboratorio (di chip pioppo). </li><li> Ottenere un pennello, paletta, biossido di cloro e alcool isopropilico (70%) per pulire l&#39;apparecchio tra le sessioni e controllo degli odori. </li></ol></li><li> Acclimatazione Apparato e il comportamento sociale <ol><li> Per questa fase dell&#39;esperimento, effettuare misurazioni in ratti adulti che sono almeno 90 giorni di età. </li><li> Prima di ogni sessione di rimuovere eventuali trucioli boschi, pulire l&#39;apparecchio pulito con alcool isopropilico per controllare gli odori tra le sessioni e fornire cippato fresco. Assicurarsi che i trucioli coprono interamente il fondo dell&#39;apparecchio. Pulire e disinfettare l&#39;apparato con un agente appropriato, ad esempio il biossido di cloro, come necessario. </li><li> Per tre giorni consecutivi inserire un suo compagno di gabbia degli animali e nellacamera per 30 minuti per acclimatare animali all&#39;apparecchio. Durante le sessioni di acclimatazione tutte le luci delle camere sono spente. </li><li> Alla fine della terza sessione di acclimatazione casa gli animali singolarmente in nuove gabbie con biancheria fresca, cibo e acqua per 24 ore per motivare l&#39;interazione sociale. </li><li> Record interazione sociale il giorno 24 ore dopo gli animali sono stati separati. <ol><li> Rimuovere i trucioli di legno dall&#39;apparecchio, pulito e disinfettare con biossido di cloro, pulire con alcool isopropilico per controllare gli odori, e sostituire i trucioli di legno prima della sessione. </li><li> Posizionare una o più telecamere per registrare l&#39;interazione. Posizione almeno una telecamera di fronte dell&#39;apparecchiatura in modo che gli specchi sulla parete posteriore dell&#39;apparecchio può fornire un&#39;ulteriore prospettiva per l&#39;analisi. </li><li> Recupero animali uno alla volta e tenere l&#39;animale di fronte alla telecamera in modo che le caratteristiche uniche della pelliccia modello possono notare. Questi elementi di identificazione diil pelo può essere utilizzata per distinguere ratti durante l&#39;analisi anziché marcatura artificialmente gli animali. NOTA: Poiché molti aspetti del roditore interazione sociale coinvolgono i segnali olfattivi e odorare il partner, ovunque devono essere evitati eventuali odori estranei introduzione. Ratti lunghi-Evans hanno tipicamente qualche caratteristica del modello di pellicce che può essere utilizzata per distinguere una data coppia di animali. Per altri ceppi (ad esempio, ratti Sprague-Dawley) potrebbero essere utilizzati approcci alternativi come la marcatura della coda con un colorante inodore. E &#39;importante riconoscere che molti comportamenti di interesse sono diretti verso obiettivi specifici (ad es., Anogenitale sniffing diretto vicino alla base della coda, attacchi giocose diretti alla nuca, o attacchi aggressivi diretti verso i fianchi o la pancia). Marcatura gli animali più vicino alla punta della coda lontano da questi bersagli di interesse è raccomandato. </li><li> Registrazione video della interazione sociale per almeno 12 min.</li><li> Monitorare gli animali per combattere per tutta la sessione. Se possibile, osservare gli animali tramite un monitor o finestra in modo che lo sperimentatore non è nella camera durante la sessione. NOTA: Fighting è stato solo raramente osservata in studi con ratti adulti, invece, gli animali devono essere monitorati per tutta la sessione. La sessione dovrebbe essere cessato se non vi è lotta eccessiva o vi sono segni di danni o lesioni ad un animale. </li></ol></li></ol></li><li> Codifica comportamentale e analisi. <ol><li> Identificare i seguenti comportamenti di interesse di cui al precedente lavoro con PAE 1,18. Quantificare la durata, la frequenza e la latenza di prima occorrenza dei seguenti comportamenti; wrestling (compresi pinning), pugilato, strisciando (incrocio) sopra / sotto il partner, anogenitale sniffing, altro sniffing del corpo del partner (corpo sniffing), allogrooming (grooming del partner), l&#39;allevamento, e sniffing / scavando nei trucioli di legno . Esempi di ogni comportamento sono illustrared nel componente video di questo articolo. </li><li> Quantificare i comportamenti sociali di interesse dal video. Ottenere la frequenza, la durata totale e la latenza di prima istanza per ogni comportamento di interesse. NOTA: Ottenere queste misure può essere realizzato manualmente, tuttavia, si raccomanda di quantificazione con analisi computerizzate di video digitalizzato. A Matlab (www.mathworks.com) script per la riproduzione del video e la quantificazione dei comportamenti è fornita come supplemento a questo articolo. </li><li> Dopo codifica completata per tutti i ratti i dati relativi alla durata, frequenza e latenza risultanti vengono analizzati con un pacchetto statistico. </li></ol></li></ol>

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D., 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=Effects+of+a+Novel+Cognition-Enhancing+Agent+on+Fetal+Ethanol-Induced+Learning+Deficits.">Effects of a Novel Cognition-Enhancing Agent on Fetal Ethanol-Induced Learning Deficits.</a> Alcohol. Clin. Exp. Res. 34 (10), 1793-1802 (2010).</li><li>Goodlett, C. R., Johnson, T. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neonatal+binge+ethanol+exposure+using+intubation:+Timing+and+dose+effects+on+place+learning.">Neonatal binge ethanol exposure using intubation: Timing and dose effects on place learning.</a> Neurotoxicol. Teratol. 19 (6), 435-446 (1997).</li><li>Staples, M. C., Rosenberg, M. J., Allen, N. A., Porch, M. W., Savage, D. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Impact of Combined Prenatal Ethanol and Prenatal Stress Exposure on Anxiety and Hippocampal-Sensitive Learning in Adult. 37 (12), 2039-2047 (2013).</li><li>Champagne, F. A., Francis, D. D., Mar, A., Meaney, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Variations+in+maternal+care+in+the+rat+as+a+mediating+influence+for+the+effects+of+environment+on+development.">Variations in maternal care in the rat as a mediating influence for the effects of environment on development.</a> Physiol. Behav. 79 (3), 359-371 (2003).</li><li>Hamilton, D. A., 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=Patterns+of+social-experience-related+c-fos+and+Arc+expression+in+the+frontal+cortices+of+rats+exposed+to+saccharin+or+moderate+levels+of+ethanol+during+prenatal+brain+development.">Patterns of social-experience-related c-fos and Arc expression in the frontal cortices of rats exposed to saccharin or moderate levels of ethanol during prenatal brain development.</a> Behav. Brain Res. 214 (1), 66-74 (2010).</li><li>Barnett, S. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> A study in behaviour: Principles of ethology and behavioural physiology displayed mainly in the rat. , (1963).</li><li>Pellis, S. M., Pellis, V. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Play-fighting+differs+from+serious+fighting+in+both+target+of+attack+and+tactics+of+fighting+in+the+laboratory+rat+Rattus-Norvegicus.">Play-fighting differs from serious fighting in both target of attack and tactics of fighting in the laboratory rat Rattus-Norvegicus.</a> Aggressive Behav. 13 (4), 227-242 (1987).</li><li>Meaney, M. J., Stewart, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+descriptive+study+of+social-development+in+the+rat+(Rattus-Norvegicus).">A descriptive study of social-development in the rat (Rattus-Norvegicus).</a> Anim. Behav. 29 (1), 34-45 (1981).</li><li>Pellis, S. M., Pellis, V. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+prejuvenile+onset+of+play+fighting+in+laboratory+rats+(Rattus+norvegicus).">The prejuvenile onset of play fighting in laboratory rats (Rattus norvegicus).</a> Dev. Psychobiol. 31 (3), 193-205 (1997).</li><li>Himmler, B. T., Pellis, V. C., Pellis, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Peering+into+the+dynamics+of+social+interactions:+measuring+play+fighting+in+rats.">Peering into the dynamics of social interactions: measuring play fighting in rats.</a> J. Vis. Exp. (71), e4288-e4288 (2013).</li><li>Panksepp, J., Siviy, S., Normansell, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+psychobiology+of+play+:+Theoretical+and+methodological+perspectives.">The psychobiology of play : Theoretical and methodological perspectives.</a> Neurosci. Biobehav. Rev. 8 (4), 465-492 (1984).</li><li>Siviy, S. M., Panksepp, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sensory+modulation+of+juvenile+play+in+rats.">Sensory modulation of juvenile play in rats.</a> Dev. Psychobiol. 20 (1), 39-55 (1987).</li><li>Pellis, S. M., 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+effects+of+orbital+frontal+cortex+damage+on+the+modulation+of+defensive+responses+by+rats+in+playful+and+nonplayful+social+contexts.">The effects of orbital frontal cortex damage on the modulation of defensive responses by rats in playful and nonplayful social contexts.</a> Behav. Neurosci. 120 (1), 72-84 (2006).</li><li>Blanchard, R. J., Blanchard, D. C., Takahashi, T., Kelley, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Attack+and+defensive+behavior+in+albino-rat.">Attack and defensive behavior in albino-rat.</a> Anim. Behav. 25 (3), 622-634 (1977).</li></ol>

The authors have no conflicts of interest to disclose.

Il paradigma esposizione prenatale qui descritto comporta il consumo volontario di etanolo (5% v / v) di dighe ratto durante la gravidanza. Ci sono un certo numero di protocolli per esporre gli animali non umani a etanolo durante lo sviluppo cerebrale rappresentato in letteratura, che differiscono rispetto alla temporizzazione, dose, la durata e la via di somministrazione etanolo nonché specie in esame. Anche se uno studio approfondito dei vantaggi dei vari protocolli di esposizione non è fornito qui, numerosi vantagg...

Si stima che circa 1-5% dei bambini sono diagnosticati con Fetal Alcohol Spectrum Disorders (FASDs) 2, che comprendono la sindrome alcolica fetale (FAS), parziale FAS (PFA), e alcolcorrelati dello sviluppo neurologico Disturbi (ARNDs) 3. I deficit nel comportamento sociale e cognizione sono tra gli esiti avversi più comuni osservati nei bambini con FASDs 4-7. Conseguenze negative non sono limitati a forte esposizione prenatale (PAE), come moderato PAE che non porta al deficit morfologici, comportamentali e cognitivi cospicua caratteristica del FAS può causare relativamente sottili, ma comunque persistente, deficit negli esseri umani con FASDs 8-10 e gli animali non umani esposti all&#39;etanolo durante lo sviluppo del cervello 11. L&#39;importanza di comprendere le conseguenze neurobiologiche comportamentali e corrispondenti di moderata PAE è sottolineata da stime attuali indicano che la grande maggioranza dei casi FASD rientrano fra le meno sgamma Evere dello spettro 12. Diversi laboratori indipendenti hanno segnalato alterazioni roditore comportamento sociale legati a etanolo esposizione durante lo sviluppo del cervello, tra cui una diminuzione di indagine e di interazione 1,13-15, alterata gioco 14,16,17, aumento interazioni aggressive 17,18, alterazioni nella risposta a stimoli sociali 19-21, e deficit in preferenze alimentari socialmente acquisite e la memoria di riconoscimento sociale 22. Deficit comportamento sociale sono stati osservati a seguito dell&#39;esposizione ad pesanti (concentrazioni di etanolo nel sangue (BECS) ~ 300 mg / dl) 22,23 o livelli più moderati di etanolo (BEC ~ 80mg / dl) 1, e attraverso una vasta gamma di parametri per altro significativo fattori, tra cui i tempi di esposizione, durata dell&#39;esposizione, e l&#39;età al momento della valutazione del comportamento. Precedenti ricerche hanno dimostrato che le alterazioni in aspetti specifici di int socialeeraction in età adulta discriminare ratti esposti a livelli moderati di alcol da animali di controllo esposti a saccarina 1,18. In particolare, moderata PAE ha sempre stato associato con aumenti robusti di wrestling, che suggerisce un aumento del comportamento aggressivo, e più bassi livelli di indagine sociale (ad esempio, fiutando del partner) in età adulta. Perché alterazioni nel comportamento sociale sono conseguenze affidabili PAE, la quantificazione del comportamento sociale in seguito PAE può tenere importanza per progredire verso una più completa comprensione delle basi neurali di alterazioni PAE-correlate nel comportamento sociale e lo sviluppo di approcci interventistici. L&#39;obiettivo di questo documento e il video associato è di fornire istruzioni sul protocollo PAE moderato e metodi per la quantificazione del comportamento sociale in figli adulti che hanno affidabile distinto prenatale alcol esposti da prole non esposti rat.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Saccharin sodium salt hydrate</td>
			<td>Sigma</td>
			<td>S1002</td>
			<td></td>
		</tr>
		<tr>
			<td>190 proof ethanol</td>
			<td>Sigma</td>
			<td>493538</td>
			<td></td>
		</tr>
		<tr>
			<td>Beaded glass drinking tubes</td>
			<td>Fisher</td>
			<td>14-955K</td>
			<td></td>
		</tr>
		<tr>
			<td>Natural rubber white #4 stopper one hole</td>
			<td>Plasticoid</td>
			<td>LSG4M181</td>
			<td></td>
		</tr>
		<tr>
			<td>1&#34; bend tubes-ball point</td>
			<td>Ancare</td>
			<td>TD-199-3&#34;</td>
			<td></td>
		</tr>
		<tr>
			<td>Paper rulers</td>
			<td>N/A</td>
			<td>N/A</td>
			<td><a href="http://www.vendian.org/mncharity/dir3/paper_rulers">www.vendian.org/mncharity/dir3/paper_rulers</a></td>
		</tr>
		<tr>
			<td>Apparatus for social interaction</td>
			<td>Custom built</td>
			<td>N/A</td>
			<td>95 cm X 47 cm X 43 cm</td>
		</tr>
		<tr>
			<td>Video cameras</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>Capable of recording low/no light conditions</td>
		</tr>
		<tr>
			<td>Infrared illuminators</td>
			<td>Vitek</td>
			<td>VT-IR1-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Teklad laboratory grade sani-chips</td>
			<td>Harlan</td>
			<td>7090A</td>
			<td></td>
		</tr>
		<tr>
			<td>Brush and dustpan</td>
			<td>N/A</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr>
			<td>Isopropyl alcohol</td>
			<td>Sigma</td>
			<td>W292907</td>
			<td></td>
		</tr>
		<tr>
			<td>Chlorine Dioxide (1.5 mg Tablets)</td>
			<td>Quiplabs</td>
			<td>N/A</td>
			<td>Prepare per manufacturer&#39;s recommendation</td>
		</tr>
	</tbody>
</table>

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.

Nel corso di molti cicli di allevamento ratti femmina nella condizione etanolo bere costantemente una media di circa 2,1 g / kg di etanolo per 4 hr bevuta. Dighe Rat consumano circa la metà del totale di quattro ore durante il primo 15 a 30 minuti dopo l&#39;introduzione dei tubi bere, causando un picco di concentrazione di etanolo nel siero materno di circa 60 mg / dl, misurata al punto di tempo di 45 min. Il restante 3,5 ore del periodo di bere, continuano a consumare 5% di etanolo a un tasso inferiore, ma relativame...

Watch this Scientific Journal Video about Moderate Prenatal Alcohol Exposure and Quantification of Social Behavior in Adult Rats at JoVE.com

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

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 ...

moderate-prenatal-alcohol-exposure-quantification-social-behavior

Research

JoVE Journal

Behavior

9.6K Views.  University of New Mexico. 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. 

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

A Procedure to Observe Context-induced Renewal of Pavlovian-conditioned Alcohol-seeking Behavior in Rats

Environmental contexts in which drugs of abuse are consumed can trigger craving, a subjective Pavlovian-conditioned response that can facilitate drug-seeking behavior and prompt relapse in abstinent drug users. We have developed a procedure to study the behavioral and neural processes that mediate the impact of context on alcohol-seeking behavior in rats. Following acclimation to the taste and pharmacological effects of 15% ethanol in the home cage, male Long-Evans rats receive Pavlovian discrimination training (PDT) in conditioning chambers. In each daily (Mon-Fri) PDT session, 16 trials each of two different 10 sec auditory conditioned stimuli occur. During one stimulus, the CS+, 0.2 ml of 15% ethanol is delivered into a fluid port for oral consumption. The second stimulus, the CS-, is not paired with ethanol. Across sessions, entries into the fluid port during the CS+ increase, whereas entries during the CS- stabilize at a lower level, indicating that a predictive association between the CS+ and ethanol is acquired. During PDT each chamber is equipped with a specific configuration of visual, olfactory and tactile contextual stimuli. Following PDT, extinction training is conducted in the same chamber that is now equipped with a different configuration of contextual stimuli. The CS+ and CS- are presented as before, but ethanol is withheld, which causes a gradual decline in port entries during the CS+. At test, rats are placed back into the PDT context and presented with the CS+ and CS- as before, but without ethanol. This manipulation triggers a robust and selective increase in the number of port entries made during the alcohol predictive CS+, with no change in responding during the CS-. This effect, referred to as context-induced renewal, illustrates the powerful capacity of contexts associated with alcohol consumption to stimulate alcohol-seeking behavior in response to Pavlovian alcohol cues.

A procedure to study the capacity of an alcohol associated environmental context to trigger the renewal of alcohol-seeking behavior in rats is described.

Una Procedura di osservare Context-indotta Rinnovo pavloviano condizionata alcol-seeking comportamento in Rats

Environmental contexts in which drugs of abuse are consumed can trigger craving, a subjective Pavlovian-conditioned response that can facilitate ...

Ricerca

Comportamento

The Modified Hole Board - Measuring Behavior, Cognition and Social Interaction in Mice and Rats

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure multiple dimensions of unconditioned behavior in small laboratory mammals (e.g., mice, rats, tree shrews and small primates). This paradigm is a valuable alternative for the use of a behavioral test battery, since a broad behavioral spectrum of an animal&#8217;s behavioral profile can be investigated in one single test.
The apparatus consists of a box, representing the &#8216;protected&#8217; area, separated from a group compartment. A board, on which small cylinders are staggered in three lines, is placed in the center of the box, representing the &#8216;unprotected&#8217; area of the set-up. The cognitive abilities of the animals can be measured by baiting some cylinders on the board and measuring the working and reference memory. Other unconditioned behavior, such as activity-related-, anxiety-related- and social behavior, can be observed using this paradigm. Behavioral flexibility and the ability to habituate to a novel environment can additionally be observed by subjecting the animals to multiple trials in the mHB, revealing insight into the animals&#8217; adaptive capacities.
Due to testing order effects in a behavioral test battery, na&#239;ve animals should be used for each individual experiment. By testing multiple behavioral dimensions in a single paradigm and thereby circumventing this issue, the number of experimental animals used is reduced. Furthermore, by avoiding social isolation during testing and without the need to food deprive the animals, the mHB represents a behavioral test system, inducing if any, very low amount of stress.

This protocol describes the modified hole board, which is a behavioral test set-up that comprises the characteristics of an open field and a traditional hole board. This set-up enables the differential analysis of unconditioned behavior of small laboratory mammals as well as the analysis of cognitive abilities.

The Hole Consiglio Modified - Behavior, Cognition e l&#39;interazione sociale in topi e ratti di misura

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure ...

Rodent Brain Microinjection to Study Molecular Substrates of Motivated Behavior

Brain microinjection can aid elucidation of the molecular substrates of complex behaviors, such as motivation. For this purpose rodents can serve as appropriate models, partly because the response to behaviorally relevant stimuli and the circuitry parsing stimulus-action outcomes is astonishingly similar between humans and rodents. In studying molecular substrates of complex behaviors, the microinjection of reagents that modify, augment, or silence specific systems is an invaluable technique. However, it is crucial that the microinjection site is precisely targeted in order to aid interpretation of the results. We present a method for the manufacture of surgical implements and microinjection needles that enables accurate microinjection and unlimited customizability with minimal cost. Importantly, this technique can be successfully completed in awake rodents if conducted in conjunction with other JoVE articles that covered requisite surgical procedures. Additionally, there are many behavioral paradigms that are well suited for measuring motivation. The progressive ratio is a commonly used method that quantifies the efficacy of a reinforcer to maintain responding despite an (often exponentially) increasing work requirement. This assay is sensitive to reinforcer magnitude and pharmacological manipulations, which allows reinforcing efficacy and/ or motivation to be determined. We also present a straightforward approach to program operant software to accommodate a progressive ratio reinforcement schedule.

Rodents are an appropriate model to investigate the molecular substrates of behavior and complex psychiatric disorders. Brain microinjection in awake rodents can be used to elucidate disease substrates. An efficient and customizable brain microinjection method as well as the execution of an operant paradigm that quantifies motivation is presented.

Roditore cervello microiniezione di studio molecolare substrati di Motivato Behavior

Brain microinjection can aid elucidation of the molecular substrates of complex behaviors, such as motivation. For this purpose rodents can serve as ...

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 ...

A Method for Evaluating the Reinforcing Properties of Ethanol in Rats without Water Deprivation, Saccharin Fading or Extended Access Training

Operant oral self-administration methods are commonly used to study the reinforcing properties of ethanol in animals. However, the standard methods require saccharin/sucrose fading, water deprivation and/or extended training to initiate operant responding in rats. This paper describes a novel and efficient method to quickly initiate operant responding for ethanol that is convenient for experimenters and does not require water deprivation or saccharin/sucrose fading, thus eliminating the potential confound of using sweeteners in ethanol operant self-administration studies. With this method, Wistar rats typically acquire and maintain self-administration of a 20% ethanol solution in less than two weeks of training. Furthermore, blood ethanol concentrations and rewards are positively correlated for a 30 min self-administration session. Moreover, naltrexone, an FDA-approved medication for alcohol dependence that has been shown to suppress ethanol self-administration in rodents, dose-dependently decreases alcohol intake and motivation to consume alcohol for rats self-administering 20% ethanol, thus validating the use of this new method to study the reinforcing properties of alcohol in rats.

This protocol describes a novel and efficient method to quickly initiate operant responding for ethanol in rats that, contrary to standard methods, does not require water deprivation or saccharin/sucrose fading to initiate responding.

Un metodo per valutare le proprietà di rinforzo di etanolo in ratti senza acqua privazione, Saccarina dissolvenza o di formazione accesso esteso

Operant oral self-administration methods are commonly used to study the reinforcing properties of ethanol in animals. However, the standard methods ...

Examining Recall Memory in Infancy and Early Childhood Using the Elicited Imitation Paradigm

The ability to recall the past allows us to report on details of previous experiences, from the everyday to the significant. Because recall memory is commonly assessed using verbal report paradigms in adults, studying the development of this ability in preverbal infants and children proved challenging. Over the past 30 years, researchers have developed a non-verbal means of assessing recall memory known as the elicited or deferred imitation paradigm. In one variant of the procedure, participants are presented with novel three-dimensional stimuli for a brief baseline period before a researcher demonstrates a series of actions that culminate in an end- or goal-state. The participant is allowed to imitate the demonstrated actions immediately, after a delay, or both. Recall performance is then compared to baseline or to performance on novel control sequences presented at the same session; memory can be assessed for the individual target actions and the order in which they were completed. This procedure is an accepted analogue to the verbal report techniques used with adults, and it has served to establish a solid foundation of the nature of recall memory in infancy and early childhood. In addition, the elicited or deferred imitation procedure has been modified and adapted to answer questions relevant to other aspects of cognitive functioning. The broad utility and application of imitation paradigms is discussed, along with limitations of the approach and directions for future research.

The elicited imitation procedure was established to examine the development of recall memory in infancy and early childhood. This procedure has been widely used to establish a solid foundation of the nature of recall memory in infancy and early childhood.

Esaminando Richiamo Memoria in prima e seconda infanzia Utilizzando il suscitato imitazione Paradigma

The ability to recall the past allows us to report on details of previous experiences, from the everyday to the significant. Because recall memory is ...

The Rodent Psychomotor Vigilance Test (rPVT): A Method for Assessing Neurobehavioral Performance in Rats and Mice

The human Psychomotor Vigilance Test (PVT) is a widely used procedure for measuring changes in fatigue and sustained attention. The present article describes a rodent version of the PVT&#8212;termed the &#34;rPVT&#34;&#8212;that measures similar aspects of attention (i.e., performance accuracy, motor speed, premature responding, and lapses in attention). Data are presented that demonstrate both the short- and long-term usefulness of the rPVT when employed with laboratory rats. Rats easily learn the rPVT, and learning to perform the basic procedure takes less than two weeks of training. Once acquired, rat performances in the rPVT show a high degree of similarity to these same performance measures in the human PVT, including similarities in, lapses in attention, reaction times, vigilance decrements across session time (i.e., the human &#34;time-on-task&#34; effects), and the response-stimulus interval (RSI) effect described for humans. Thus the rPVT can be an extremely valuable tool for assessing the effects of a wide range of variables on sustained attention quite similar to human PVT performances, and thus can be useful for developing novel treatments for neurobehavioral dysfunctions.

The Rodent Psychomotor Vigilance Test rPVT: A Method for Assessing Neurobehavioral Performance in Rats and Mice

A rat version of the human Psychomotor Vigilance Test (PVT) is described that measures aspects of attention similar to those measured with the human PVT, including aspects of human vigilance such as performance accuracy, motor speed, premature responding, and lapses in attention.

Il roditore psicomotorio vigilanza Test (rPVT): un metodo per la valutazione delle prestazioni neurocomportamentali in ratti e topi

The human Psychomotor Vigilance Test (PVT) is a widely used procedure for measuring changes in fatigue and sustained attention. The present article ...

Using Clicker Training and Social Observation to Teach Rats to Voluntarily Change Cages

Cage cleaning is a routinely performed husbandry procedure and is known to induce stress in laboratory rats. As stress can have a negative impact on well-being and can affect the comparability and reproducibility of research results, the amount of stress experienced by laboratory animals should be minimized and avoided when possible. Further, the direct contact between the rat and animal caretaker during the cage change bears hygiene risks and therefore possibly negatively impacts the well-being of the rats and the quality of the research. 
Our protocol aims to improve the routinely performed cage changing procedure. For this reason, we present a feasible protocol that enables rats to learn via clicker training and observation to voluntarily change to a clean cage. This training helps to reduce stress caused by the physical disturbance and handling associated with the cage changes and concurrently enables a reduction in direct contact between animal and animal caretaker after the training phase is completed.
The implementation of clicker training to rats is fast and easy. Rats are generally interested in the training and efficiently learn the desired behavior, which entails changing cages through a pipe. Even without training, the rats learn to perform the desired behavior by observation, as 80% of the observational learning group successfully changed cages when tested. The training further helps to establish a relationship of trust between trainer and animal. As hygiene and well-being are both very important in animal experiments, this protocol might also help to improve high-quality research.

This protocol introduces a method of cage change for rats via clicker training. Rats learn the desired behavior not only by direct training but also by observational learning. The implementation of this fast and easy protocol might help to improve well-being and hygiene in rodent facilities.

Utilizzando Clicker Training e osservazione sociale per insegnare ratti volontariamente cambiare gabbie

Cage cleaning is a routinely performed husbandry procedure and is known to induce stress in laboratory rats. As stress can have a negative impact on ...

Integrating Computerized Linguistic and Social Network Analyses to Capture Addiction Recovery Capital in an Online Community

The article describes a new methodology designed with the aim of finding a comprehensive, unobtrusive, and accurate way of capturing social recovery capital development in online communities of recovery from alcohol and drug (AOD) addiction. Recovery capital was conceptualised as both engagement in the online recovery community and identification with the community. To measure recovery capital development, naturally occurring data were extracted from the social media page of a specific recovery program, with the page being set up as a resource for a face-to-face recovery program. To map engagement with the online community, social network analysis (SNA) capturing online social interaction was performed. Social interaction was measured through the linkages between the online contributors/members of the online community as represented by program clients, staff, and supporters from the broader community. To capture markers of social identification with the online community, computerised linguistic analysis of the textual data (content from posts and comments) was conducted. Recovery capital captured in this way was analysed against retention data (a proxy outcome indicator), as days spent in the (face-to-face) recovery program. The online data extracted was linked to participant data in regards to program retention to test prediction of a key recovery outcome. This approach allowed the examination of the role of online support communities and assessment of the association between recovery capital (developed via the online community of recovery) and recovery outcomes.

The article describes a novel approach for analyzing dynamic online social interactions (in an online context) exemplified by a study of an online community of recovery from alcohol and drug addiction.

Integrazione di analisi linguistiche e sociali computerizzate per catturare il capitale di recupero dalle dipendenze in una comunità online

The article describes a new methodology designed with the aim of finding a comprehensive, unobtrusive, and accurate way of capturing social recovery ...

A Complex Diving-For-Food Task to Investigate Social Organization and Interactions in Rats

For many species, where status is a vital motivator that can affect health, social hierarchies influence behavior. Social hierarchies that include dominant-submissive relationships are common in both animal and human societies. These relationships can be affected by interactions with others and with their environment, making them difficult to analyze in a controlled study. Rather than a simple dominance hierarchy, this formation has a complicated presentation that allows rats to avoid aggression. Status can be stagnant or mutable, and results in complex societal stratifications. Here we describe a complex diving-for-food task to investigate rodent social hierarchy and behavioral interactions. This animal model may allow us to assess the relationship between a wide range of mental illnesses and social organization, as well as to study the effectiveness of therapy on social dysfunction.

This protocol describes a method of examining social hierarchy in a rat model. Rats perform a complex diving-for-food task in which they form a distinct hierarchy according to their willingness to dive underwater and swim to obtain a food pellet. This method is used to understand decision making and social relationships among highly social animals in small groups.