Questo lavoro è supportato da una concessione Research Council Baylor University e da un assegno di ricerca della Fondazione Epilessia.

I topi utilizzati nel seguente esperimento sono stati generati e ospitati presso la Baylor University a una temperatura ambiente di 22 ° C, con una luce 14 ore e 10 ore scuro (20:00-06:00 h) ciclo diurno. I topi sono stati dati ad libitum l&#39;accesso al cibo e all&#39;acqua. Tutte le procedure per i topi erano in conformità con il National Institutes of Linee guida per la salute per la cura e l&#39;uso di animali da laboratorio e il protocollo animale è stato approvato dal Baylor University cura degli animali e del Comitato Usa. Panoramica Il compito paura traccia condizionata, basato su procedure descritte da Wiltgen e colleghi 9. 1. Preparazione delle attrezzature Il timore camera di un&#39;apparecchiatura di condizionamento (26 centimetri x 22 cm x 18 cm) costituito da due lati che sono acrilico, due lati che sono metallo ed estremità reticolati che viene utilizzato per erogare la scarica piede lieve. La camera di prova è la casad in un suono da camera attenuato. La camera è anche stretto luce per evitare che la luce esterna da influenzare il software di rilevamento del movimento. <ol><li> Calibrare i livelli di shock, livelli di luce, e livelli di intensità del suono per la camera di prova. Misurare il livello della camera di prova sfondo. Il rumore di fondo in questa camera è di 65 dB. Utilizzare un fonometro per misurare questo livello. Il contatore deve essere impostato a 70 dB, impostato su C, e al rilevamento impostazione lento. <ol><li> Calibrare il livello di shock a 0,5 mA. Utilizzare un dispositivo di calibrazione esterno per calibrare correttamente il livello di shock (vedi Materiali Tavolo). La misura interna del generatore scossa non è preciso. Il generatore di shock amministra uno shock criptato che non può essere misurata con precisione da un amperometro standard. </li><li> Collocare un cavo su una delle sbarre di griglia e posizionare l&#39;altro cavo 3 o 4 bar sopra. Utilizzare il generatore di shock per somministrare lo shock. Regolare il livello di shock fino a quando il livello corretto è raggiunto. Fate questo per EAch temere camera di condizionamento. </li><li> Chiudere la porta alla camera quando si misura il livello di luminosità della camera. Calibrare livello la luce a circa 1,0. Questo è un numero specifico per il software FreezeFrame utilizzato in questo esperimento. Un esposimetro esterno leggerà questo come 2 lux. L&#39;intensità luminosa può essere regolata spostando la posizione della luce casa o regolando il condensatore della lente. Assicurarsi di stringere le viti di regolazione per la lente dopo la regolazione. </li><li> Calibrare il livello del suono di decibel a 85 dB. Utilizzare un misuratore di decibel esterno dentro la gabbia di test per calibrare il livello di decibel (vedi Materiali Tavolo). Il suono ha presentato sarà un tono di 2.700 Hz. Nota: Se si utilizzano topi di età superiore ai 6 mesi di età, può essere meglio usare rumore bianco dal momento che i topi più anziani possono avere deficit dell&#39;udito. </li></ol></li><li> Dopo che l&#39;apparecchio è pronto prendere i soggetti in una stanza un&#39;azienda distinta. Nota: non ospitare il mouse nella stessa stanza come sala prove. </li><li> Label the coda dei topi che saranno testati in ordine delle prove. E &#39;meglio ridurre le eccessive manipolazioni prima del test. In alternativa, le code possono essere etichettati il ​​giorno prima dell&#39;esperimento per ridurre lo stress da maneggiamento. Dopo i topi sono stati etichettati permettono loro di acclimatarsi alla camera per 30 min. Avere gabbie in più pulite per alloggiare i topi dopo il test è stato completato. </li></ol> 2. Traccia condizionata Day 1 <ol><li> Rimuovere ogni mouse dalla gabbia e metterli in gabbie individuali per il trasporto alla paura camera di condizionamento. Utilizzare biancheria pulita per ogni gabbia. Mettere una nota adesiva sulla gabbia di trasferimento per mantenere l&#39;ordine dei topi testare il corretto. Nota: se i topi sono alloggiati singolarmente quindi possono essere trasportati nella loro gabbia casa. </li><li> Posizionare il mouse nella camera di prova e chiudere la porta. Avviare il programma software. </li><li> Il giorno di formazione, consentono ai topi di esplorare la camera per 3 min. Il software presenta poi un secondo tono (85 dB, 2.700 Hz) t 20o l&#39;animale. Dopo un periodo di traccia 20 sec una leggera scossa (2 sec, 0,5 mA) viene somministrato all&#39;animale. </li><li> Registrare la reazione del soggetto per confermare che hanno ricevuto i stimoli avversi visualizzando il video. Un intervallo inter-trial 200 sec separa le 5 prove di condizionamento. Ogni prova è costituito da un tono 20 secondi seguita da un ritardo di 20 sec poi uno shock. </li><li> Al termine del test consentire agli animali di rimanere nella camera di prova per 1 min prima di rimuoverla dalla gabbia test. </li><li> Posizionare l&#39;animale torna nella gabbia trasferimento e tornare alla sua gabbia casa. Se ci sono i topi aggiuntivi nella loro gabbia casa poi ospitare singolarmente il mouse fino a quando tutti i test completo topi. Ciò consentirà di ridurre lo stress all&#39;altra topi che non sono stati testati. Una soluzione alternativa potrebbe essere quella di alloggiare singolarmente tutti i topi una settimana prima del test per ridurre l&#39;influenza di rimuovere in continuo topi da una gabbia. </li><li> Pulire la camera di prova con il 30% di isopropanolo dopo each animale è testato. </li><li> Ripetere i passaggi 2,2-2,7 per tutti i mouse nella coorte di test. </li><li> Rientro tutti i topi nella loro stanza colonia dopo l&#39;ultimo mouse nella coorte è stato testato. </li></ol> 3. Trace paura condizionata Day 2: Prove di memoria Trace <ol><li> Test sulla memoria Trace avverrà il giorno 2. In questo protocollo ci sono 3 presentazioni di tono. Posizionare il mouse in un nuovo contesto per la prova traccia condizionata. </li><li> Preparare il software per eseguire un programma con un periodo di riferimento 2 min seguita da tre presentazioni del 20 tono sec. C&#39;è sa 220 sec ITI tra ogni presentazione di tono. </li><li> Per il nuovo condizione di contesto, posizionare inserti acriliche chiare sul pavimento della camera a modificare la forma, la consistenza e il colore della camera di condizionamento. </li><li> Cambiare l&#39;odore nella camera posizionando l&#39;estratto di vaniglia in una barca pesare sotto l&#39;inserto pavimento. </li><li> Pulire la camera con il 70% di etanolo invece del 30% di isopropanolo. Nota: Questo aiuteràper creare un contesto romanzo. </li><li> Portare i topi nella stanza detenzione e relabel loro code per la prova, se necessario. </li><li> Preparare nuove gabbie di trasferimento contesto sostituendo le lenzuola con carta straccia. Nota: Ciò contribuirà a creare un ambito romanzo. </li><li> Posizionare il mouse nella camera di prova quindi avviare il programma. Pulire la camera con il 70% di etanolo dopo che il test è stato completato. </li><li> Riportare i topi per loro gabbia casa dopo la traccia di condizionamento è completata. Rientro tutti i topi nella loro stanza colonia quando tutti i topi sono stati testati. </li></ol> 4. Trace paura condizionata Giorno 3: Prove di memoria contestuale <ol><li> Il terzo giorno di condizionamento contestuale è condotta. Preparare il software per eseguire un programma per registrare il comportamento di congelamento per 8 min. </li><li> Pulire la camera con il 30% di isopropanolo prima delle prove e dopo aver testato ogni mouse. Il contesto deve essere identica a quella del giorno 1. Le gabbie di trasferimento devono essere le stesse in 1 giorno. </li><li&gt; Portare i topi nella stanza detenzione ed etichettare le loro code per il test. </li><li> Collocare ogni mouse nella camera di prova quindi avviare il programma. Pulire la camera con il 30% di isopropanolo al termine del test. </li><li> Rientro tutti i topi nella loro stanza colonia una volta completato. </li></ol>

<ol>
	<li>Powell, R. A., Honey, P. L., Symbaluk, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Introduction to learning and behavior. , .</li><li>Tsaltas, E., Preston, G. C., Gray, J. A. <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+dorsal+bundle+lesions+on+serial+and+trace+conditioning.">The effects of dorsal bundle lesions on serial and trace conditioning.</a> Behav. Brain Res. 10, 361-374 (1983).</li><li>McAlonan, G. M., Dawson, G. R., Wilkinson, L. O., Robbins, T. W., Everitt, B. J. <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+AMPA-induced+lesions+of+the+medial+septum+and+vertical+limb+nucleus+of+the+diagonal+band+of+Broca+on+spatial+delayed+non-matching+to+sample+and+spatial+learning+in+the+water+maze.">The effects of AMPA-induced lesions of the medial septum and vertical limb nucleus of the diagonal band of Broca on spatial delayed non-matching to sample and spatial learning in the water maze.</a> Eur. J. Neurosci. 7, 1034-1049 (1995).</li><li>Chowdhury, N., Quinn, J. J., Fanselow, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dorsal+hippocampus+involvement+in+trace+fear+conditioning+with+long,+but+not+short,+trace+intervals+in+mice.">Dorsal hippocampus involvement in trace fear conditioning with long, but not short, trace intervals in mice.</a> Behav. Neurosci. 119, 1396-1402 (2005).</li><li>Quinn, J. J., Oommen, S. S., Morrison, G. E., Fanselow, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Post-training+excitotoxic+lesions+of+the+dorsal+hippocampus+attenuate+forward+trace,+backward+trace,+and+delay+fear+conditioning+in+a+temporally+specific+manner.">Post-training excitotoxic lesions of the dorsal hippocampus attenuate forward trace, backward trace, and delay fear conditioning in a temporally specific manner.</a> Hippocampus. 12, 495-504 (2002).</li><li>Misane, I., 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=Time-dependent+involvement+of+the+dorsal+hippocampus+in+trace+fear+conditioning+in+mice.">Time-dependent involvement of the dorsal hippocampus in trace fear conditioning in mice.</a> Hippocampus. 15, 418-426 (2005).</li><li>Quinn, J. J., Loya, F., Ma, Q. D., Fanselow, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dorsal+hippocampus+NMDA+receptors+differentially+mediate+trace+and+contextual+fear+conditioning.">Dorsal hippocampus NMDA receptors differentially mediate trace and contextual fear conditioning.</a> Hippocampus. 15, 665-674 (2005).</li><li>McEchron, M. D., Bouwmeester, H., Tseng, W., Weiss, C., Disterhoft, 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=Hippocampectomy+disrupts+auditory+trace+fear+conditioning+and+contextual+fear+conditioning+in+the+rat.">Hippocampectomy disrupts auditory trace fear conditioning and contextual fear conditioning in the rat.</a> Hippocampus. 8, 638-646 (1998).</li><li>Wiltgen, B. J., Sanders, M. J., Ferguson, C., Homanics, G. E., Fanselow, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trace+fear+conditioning+is+enhanced+in+mice+lacking+the+delta+subunit+of+the+GABAA+receptor.">Trace fear conditioning is enhanced in mice lacking the delta subunit of the GABAA receptor.</a> Learn. Mem. 12, 327-333 (2005).</li><li>Davis, R. R., 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=Genetic+basis+for+susceptibility+to+noise-induced+hearing+loss+in+mice.">Genetic basis for susceptibility to noise-induced hearing loss in mice.</a> Hear. Res. 155, 82-90 (2001).</li><li>Zheng, Q. Y., Johnson, K. R., Erway, L. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+hearing+in+80+inbred+strains+of+mice+by+ABR+threshold+analyses.">Assessment of hearing in 80 inbred strains of mice by ABR threshold analyses.</a> Hear. Res. 130, 94-107 (1999).</li><li>Moyer, J. R., Thompson, L. T., Disterhoft, 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=Trace+eyeblink+conditioning+increases+CA1+excitability+in+a+transient+and+learning-specific+manner.">Trace eyeblink conditioning increases CA1 excitability in a transient and learning-specific manner.</a> 16, 5536-5546 (1996).</li><li>Leuner, B., Falduto, J., Shors, T. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Associative+memory+formation+increases+the+observation+of+dendritic+spines+in+the+hippocampus.">Associative memory formation increases the observation of dendritic spines in the hippocampus.</a> J. Neurosci. 23, 659-665 (2003).</li><li>McEchron, M. D., Disterhoft, 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=Hippocampal+encoding+of+non-spatial+trace+conditioning.">Hippocampal encoding of non-spatial trace conditioning.</a> Hippocampus. 9, 385-396 (1999).</li><li>McLaughlin, J., Skaggs, H., Churchwell, J., Powell, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Medial+prefrontal+cortex+and+pavlovian+conditioning:+trace+versus+delay+conditioning.">Medial prefrontal cortex and pavlovian conditioning: trace versus delay conditioning.</a> Behav. Neurosci. 116, 37-47 (2002).</li><li>Runyan, J. D., Moore, A. N., Dash, P. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+role+for+prefrontal+cortex+in+memory+storage+for+trace+fear+conditioning.">A role for prefrontal cortex in memory storage for trace fear conditioning.</a> J. Neurosci. 24, 1288-1295 (2004).</li><li>Gilmartin, M. R., McEchron, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Single+neurons+in+the+medial+prefrontal+cortex+of+the+rat+exhibit+tonic+and+phasic+coding+during+trace+fear+conditioning.">Single neurons in the medial prefrontal cortex of the rat exhibit tonic and phasic coding during trace fear conditioning.</a> Behav. Neurosci. 119, 1496-1510 (2005).</li><li>Crow, T., Xue-Bian, J. J., Siddiqi, V., Kang, Y., Neary, J. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phosphorylation+of+mitogen-activated+protein+kinase+by+one-trial+and+multi-trial+classical+conditioning.">Phosphorylation of mitogen-activated protein kinase by one-trial and multi-trial classical conditioning.</a> J. Neurosci. 18, 3480-3487 (1998).</li><li>Martin, K. C., 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=MAP+kinase+translocates+into+the+nucleus+of+the+presynaptic+cell+and+is+required+for+long-term+facilitation+in+Aplysia.">MAP kinase translocates into the nucleus of the presynaptic cell and is required for long-term facilitation in Aplysia.</a> Neuron. 18, 899-912 (1997).</li><li>Crestani, F., 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=Trace+fear+conditioning+involves+hippocampal+alpha5+GABA(A)+receptors.">Trace fear conditioning involves hippocampal alpha5 GABA(A) receptors.</a> Proc. Natl. Acad. Sci. U.S.A. 99, 8980-8985 (2002).</li><li>Crestani, F., 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=Decreased+GABAA-receptor+clustering+results+in+enhanced+anxiety+and+a+bias+for+threat+cues.">Decreased GABAA-receptor clustering results in enhanced anxiety and a bias for threat cues.</a> Nat. Neurosci. 2, 833-839 (1999).</li><li>Moore, M. 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=Trace+and+contextual+fear+conditioning+is+enhanced+in+mice+lacking+the+alpha4+subunit+of+the+GABA(A)+receptor.">Trace and contextual fear conditioning is enhanced in mice lacking the alpha4 subunit of the GABA(A) receptor.</a> Neurobiol. Learn. Mem. 93, 383-387 (2010).</li><li>Cushman, J. D., Moore, M. D., Jacobs, N. S., Olsen, R. W., Fanselow, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+pharmacogenetic+analysis+on+the+role+of+the+alpha4+GABA(A)+receptor+subunit+in+the+ethanol-mediated+impairment+of+hippocampus-dependent+contextual+learning.">Behavioral pharmacogenetic analysis on the role of the alpha4 GABA(A) receptor subunit in the ethanol-mediated impairment of hippocampus-dependent contextual learning.</a> Alcohol Clin. Exp. Res. 35, 1948-1959 (2011).</li><li>Raybuck, J. D., Lattal, K. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Double+dissociation+of+amygdala+and+hippocampal+contributions+to+trace+and+delay+fear+conditioning.">Double dissociation of amygdala and hippocampal contributions to trace and delay fear conditioning.</a> PLoS ONE. 6, (2011).</li><li>Kwapis, J. L., Jarome, T. J., Schiff, J. C., Helmstetter, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Memory+consolidation+in+both+trace+and+delay+fear+conditioning+is+disrupted+by+intra-amygdala+infusion+of+the+protein+synthesis+inhibitor+anisomycin.">Memory consolidation in both trace and delay fear conditioning is disrupted by intra-amygdala infusion of the protein synthesis inhibitor anisomycin.</a> Learn. Mem. 18, 728-732 (2011).</li><li>Gilmartin, M. R., Kwapis, J. L., Helmstetter, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trace+and+contextual+fear+conditioning+are+impaired+following+unilateral+microinjection+of+muscimol+in+the+ventral+hippocampus+or+amygdala,+but+not+the+medial+prefrontal+cortex.">Trace and contextual fear conditioning are impaired following unilateral microinjection of muscimol in the ventral hippocampus or amygdala, but not the medial prefrontal cortex.</a> Neurobiol. Learn. Mem. 97, 452-464 (2012).</li><li>Baysinger, A. N., Kent, B. A., Brown, T. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Muscarinic+receptors+in+amygdala+control+trace+fear+conditioning.">Muscarinic receptors in amygdala control trace fear conditioning.</a> PLoS ONE. 7, (2012).</li><li>Wanisch, K., Tang, J., Mederer, A., Wotjak, C. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trace+fear+conditioning+depends+on+NMDA+receptor+activation+and+protein+synthesis+within+the+dorsal+hippocampus+of+mice.">Trace fear conditioning depends on NMDA receptor activation and protein synthesis within the dorsal hippocampus of mice.</a> Behav. Brain. 157, 63-69 (2005).</li><li>Smith, D. R., Gallagher, M., Stanton, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+background+differences+and+nonassociative+effects+in+mouse+trace+fear+conditioning.">Genetic background differences and nonassociative effects in mouse trace fear conditioning.</a> Learn. Mem. 14, 597-605 (2007).</li><li>Rudy, J. W., O'Reilly, R. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contextual+fear+conditioning,+conjunctive+representations,+pattern+completion,+and+the+hippocampus.">Contextual fear conditioning, conjunctive representations, pattern completion, and the hippocampus.</a> Behav. Neurosci. 113, 867-880 (1999).</li><li>Wiltgen, B. J., Sanders, M. J., Anagnostaras, S. G., Sage, J. R., Fanselow, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Context+fear+learning+in+the+absence+of+the+hippocampus.">Context fear learning in the absence of the hippocampus.</a> J. Neurosci. 26, 5484-5491 (2006).</li><li>Reijmers, L. G., Perkins, B. L., Matsuo, N., Mayford, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Localization+of+a+stable+neural+correlate+of+associative+memory.">Localization of a stable neural correlate of associative memory.</a> Science. 317, 1230-1233 (2007).</li><li>Huerta, P. T., Sun, L. D., Wilson, M. A., Tonegawa, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Formation+of+temporal+memory+requires+NMDA+receptors+within+CA1+pyramidal+neurons.">Formation of temporal memory requires NMDA receptors within CA1 pyramidal neurons.</a> Neuron. 25, 473-480 (2000).</li><li>Jacobs, N. S., Cushman, J. D., Fanselow, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+accurate+measurement+of+fear+memory+in+Pavlovian+conditioning:+Resolving+the+baseline+issue.">The accurate measurement of fear memory in Pavlovian conditioning: Resolving the baseline issue.</a> J. Neurosci. Methods. 190, 235-239 (2010).</li></ol>

Gli autori non hanno nulla da rivelare.

Ci sono stati diversi studi che hanno chiarito i circuiti neurali che sottende traccia paura condizionata. Trace condizionamento alla paura si ritiene di coinvolgere il CA1 dell&#39;ippocampo 12-14. Vi sono anche prove che la corteccia prefrontale mediale (mPFC) gioca un ruolo importante nella traccia occhiolino condizionata 15 e il mPFC è stato trovato per essere coinvolti in traccia paura condizionata. Uno studio ha trovato che i neuroni mPFC forniscono attività sostenuta durante il periodo di ...

In condizionamento alla paura uno stimolo neutro (NS) è accoppiato con uno stimolo incondizionato aversivo (US). Il NS è normalmente un tono e diventa uno stimolo condizionato (CS), attraverso abbinamenti ripetuti con gli Stati Uniti. Il CS può quindi provocare una risposta condizionata (CR), come il congelamento, in assenza del aversive US. Una paura protocollo condizionata comunemente usato è il ritardo condizionata. In questo protocollo l&#39;insorgenza della NS e Stati Uniti è contiguo o con qualche sovrapposizione nella presentazione dello stimolo. Anche se il ritardo condizionamento alla paura è uno dei tipi più comunemente usati di temporale condizionata associativa, ci sono molti altri tipi di condizionamento associativo modalità temporali: il condizionamento simultanea, di condizionamento all&#39;indietro, e la traccia di condizionamento 1. Nella traccia di paura condizionata c&#39;è un intervallo libero da stimolo tra NS e gli Stati Uniti di alcuni secondi con un conseguente periodo di &quot;traccia&quot;. Diversi studi hanno riportato deficits in traccia paura condizionata quando le lesioni neurotossiche sono prodotte nelle strutture che l&#39;input nei ippocampo 2-5 o quando gli agenti farmacologici sono usati per bloccare la funzione del recettore nell&#39;ippocampo. Lesione ai risultati ippocampo in deficit nella traccia di condizionamento e condizionamento contestuale, ma non compromette il ritardo condizionamento alla paura 8. Ci sono molti vantaggi di utilizzare trace fear conditioning. Il protocollo condizionata paura può essere realizzato in un periodo di test di tre giorni e permette memoria ippocampo-dipendente che non è spazialmente dipendente. Tracciare paura condizionata può essere usata come test complementare al labirinto ad acqua di Morris, prova romanzo riconoscimento di oggetti, o altri test del labirinto a indagare memoria ippocampo-dipendente.

<table border="0" cellpadding="0" cellspacing="0" width="741">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">FreezeFrame</td>
			<td style="width:143px;">Coulbourn</td>
			<td style="width:119px;"></td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">30% Isopropanol</td>
			<td style="width:143px;"></td>
			<td style="width:119px;"></td>
			<td style="width:263px;">Purchase 90% isopropanol and dilute it down to 30%</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">70% Ethanol</td>
			<td style="width:143px;"></td>
			<td style="width:119px;"></td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Amp-meter</td>
			<td style="width:143px;">Med-Associates</td>
			<td style="width:119px;">ENV-420</td>
			<td style="width:263px;">Windows XP, Vista, and 7 Compatible (32-bit only)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Digital Sound Level Meter</td>
			<td style="width:143px;"></td>
			<td style="width:119px;">33-2055</td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Vanilla Extract</td>
			<td style="width:143px;">McCormick Pure Vanilla Extract</td>
			<td style="width:119px;"></td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Sticky Notes</td>
			<td style="width:143px;">Post-it</td>
			<td style="width:119px;"></td>
			<td style="width:263px;">3X3 inch</td>
		</tr>
	</tbody>
</table>

trace fear conditioning in mice

In questo esperimento presentiamo una tecnica per misurare l&#39;apprendimento e la memoria. Nel protocollo di condizionamento trace paura presentato qui ci sono cinque abbinamenti tra uno stimolo neutro e uno stimolo incondizionato. C&#39;è un periodo di traccia di 20 secondi che separa ogni prova condizionata. Sul seguente congelamento giorno viene misurata durante la presentazione dello stimolo condizionato (CS) e il periodo di analisi. Il terzo giorno è prevista una prova 8 min per misurare memoria contestuale. I risultati sono rappresentativi di topi che sono stati presentati con lo stimolo incondizionato avversivo (shock) rispetto ai topi che hanno ricevuto le presentazioni di tono, senza lo stimolo incondizionato. Trace condizionamento alla paura è stato utilizzato con successo per rilevare i deficit e miglioramenti di apprendimento e memoria sottili nei topi che non si trovano con altri metodi di condizionamento alla paura. Questo tipo di paura condizionata è ritenuta essere dipendente connessioni tra la corteccia prefrontale mediale e l&#39;ippocampo. Una polemica attuale è se questo metodo si crede di essere amigdala-indipendente. Pertanto, è necessario un altro test di condizionamento paura per esaminare apprendimento e memoria effetti amigdala-dipendenti, ad esempio attraverso il ritardo condizionamento alla paura.

Per i risultati rappresentativi presentiamo i dati di controllo C57BL/6J topi adulti che hanno ricevuto gli abbinamenti stimolo neutro con lo stimolo incondizionato (condizione di shock) rispetto ai topi che hanno ricevuto lo stimolo neutro, ma non hanno ricevuto lo stimolo incondizionato (nessuna condizione di shock). È importante eseguire questa condizione quando prima configurazione di questa prova comportamentale per determinare se il protocollo è stato eseguito correttamente. I dati d...

Watch this Scientific Journal Video about Trace Fear Conditioning in Mice at JoVE.com

Trace Fear Conditioning in Mice

Nel seguente esperimento si descrive un protocollo per la traccia di paura condizionata nei topi. Questo tipo di memoria associativa comprende un periodo di traccia che separa lo stimolo neutro e lo stimolo incondizionato.

Trace paura condizionata in topi

In this experiment we present a technique to measure learning and memory. In the trace fear conditioning protocol presented here there are five pairings ...

Research

JoVE Journal

Behavior

32.7K Views.  Baylor University.  Nel seguente esperimento si descrive un protocollo per la traccia di paura condizionata nei topi. Questo tipo di memoria associativa comprende un periodo di traccia che separa lo stimolo neutro e lo stimolo incondizionato. 

Video: Trace Fear Conditioning in Mice

Contextual and Cued Fear Conditioning Test Using a Video Analyzing System in Mice

The contextual and cued fear conditioning test is one of the behavioral tests that assesses the ability of mice to learn and remember an association between environmental cues and aversive experiences. In this test, mice are placed into a conditioning chamber and are given parings of a conditioned stimulus (an auditory cue) and an aversive unconditioned stimulus (an electric footshock). After a delay time, the mice are exposed to the same conditioning chamber and a differently shaped chamber with presentation of the auditory cue. Freezing behavior during the test is measured as an index of fear memory. To analyze the behavior automatically, we have developed a video analyzing system using the ImageFZ application software program, which is available as a free download at http://www.mouse-phenotype.org/. Here, to show the details of our protocol, we demonstrate our procedure for the contextual and cued fear conditioning test in C57BL/6J mice using the ImageFZ system. In addition, we validated our protocol and the video analyzing system performance by comparing freezing time measured by the ImageFZ system or a photobeam-based computer measurement system with that scored by a human observer. As shown in our representative results, the data obtained by ImageFZ were similar to those analyzed by a human observer, indicating that the behavioral analysis using the ImageFZ system is highly reliable. The present movie article provides detailed information regarding the test procedures and will promote understanding of the experimental situation.

This article presents a protocol for a contextual and cued fear conditioning test using a video analyzing system to assess fear learning and memory in mice.

Contestuali e Cued paura condizionata test utilizzando un sistema di video Analizzare in topi

The contextual and cued fear conditioning test is one of the behavioral tests that assesses the ability of mice to learn and remember an association ...

Ricerca

Comportamento

Disrupting Reconsolidation of Fear Memory in Humans by a Noradrenergic &#946;-Blocker

The basic design used in our human fear-conditioning studies on disrupting reconsolidation includes testing over different phases across three consecutive days. On day 1 - the fear acquisition phase, healthy participants are exposed to a series of picture presentations. One picture stimulus (CS1+) is repeatedly paired with an aversive electric stimulus (US), resulting in the acquisition of a fear association, whereas another picture stimulus (CS2-) is never followed by an US. On day 2 - the memory reactivation phase, the participants are re-exposed to the conditioned stimulus without the US (CS1-), which typically triggers a conditioned fear response. After the memory reactivation we administer an oral dose of 40 mg of propranolol HCl, a &#946;-adrenergic receptor antagonist that indirectly targets the protein synthesis required for reconsolidation by inhibiting the noradrenaline-stimulated CREB phosphorylation. On day 3 - the test phase, the participants are again exposed to the unreinforced conditioned stimuli (CS1- and CS2-) in order to measure the fear-reducing effect of the manipulation. This retention test is followed by an extinction procedure and the presentation of situational triggers to test for the return of fear. Potentiation of the eye blink startle reflex is measured as an index for conditioned fear responding. Declarative knowledge of the fear association is measured through online US expectancy ratings during each CS presentation. In contrast to extinction learning, disrupting reconsolidation targets the original fear memory thereby preventing the return of fear. Although the clinical applications are still in their infancy, disrupting reconsolidation of fear memory seems to be a promising new technique with the prospect to persistently dampen the expression of fear memory in patients suffering from anxiety disorders and other psychiatric disorders.

Disrupting reconsolidation is a promising approach to dampen the behavioral expression of fear memory in patients with anxiety disorders or posttraumatic stress disorder. In a series of human fear conditioning studies we showed that disrupting reconsolidation by the noradrenergic &#946;-blocker propranolol is very effective in erasing conditioned fear responding.

L&#39;interruzione Ricompattamento della Paura memoria in esseri umani da un noradrenergico β-Blocker

The basic design used in our human fear-conditioning studies on disrupting reconsolidation includes testing over different phases across three consecutive ...

Protocol for Studying Extinction of Conditioned Fear in Naturally Cycling Female Rats

Extinction of conditioned fear has been extensively studied in male rodents. Recently, there have been an increasing number of studies indicating that neural mechanisms for certain behavioral tasks and response behaviors are different in females and males. Using females in research studies can represent a challenge because of the variation of gonadal hormones during their estrous cycle. This protocol describes well-established procedures that are useful in investigating the role of estrogen in fear extinction memory consolidation in female rats. Phase of the estrous cycle and exogenous estrogen administration prior to extinction training can influence extinction recall 24 hr later. The vaginal swabbing technique for estrous phase identification described here aids the examination and manipulation of naturally cycling gonadal hormones. The use of this basic rodent model may further delineate the mechanisms by which estrogen can modulate fear extinction memory in females.

Gonadal hormones such as estrogen modulate memory formation in a number of experimental paradigms including fear extinction memory. This protocol describes a set of methods for investigating the influence of gonadal hormones specifically during extinction in naturally cycling females, including estrous cycle monitoring and exogenous estrogen administration.

Protocollo per lo studio Extinction of Fear Condizionata in Naturalmente bicicletta ratti di sesso femminile

Extinction of conditioned fear has been extensively studied in male rodents. Recently, there have been an increasing number of studies indicating that ...

Quantifying Social Motivation in Mice Using Operant Conditioning

In this protocol, social motivation is measured in mice through a pair of operant conditioning paradigms. To conduct the experiments, two-chambered shuttle boxes were equipped with two operant levers (left and right) and a food receptacle in one chamber, which was then divided from the second chamber by an automated guillotine door covered by a wire grid. Different stimulus mice, rotated across testing days, served as a social stimulus behind the wire grid, and were only visible following the opening of the guillotine door. Test mice were trained to lever press in order to open the door and gain access to the stimulus partner for 15 sec. The number of lever presses required to obtain the social reward progressively increased on a fixed schedule of 3. Testing sessions ended after test mice stopped lever pressing for 5 consecutive minutes. The last reinforced ratio or breakpoint can be used as a quantitative measure of social motivation. For the second paradigm, test mice were trained to discriminate between left and right lever presses in order to obtain either a food reward or the social reward. Mice were rewarded for every 3 presses of each respective lever. The number of food and social rewards can be compared as a measurement of the value placed upon each reward. The ratio of each reward type can also be compared between mouse strains and the change in this ratio can be monitored within testing sessions to measure satiation with a given reward type. Both of these operant conditioning paradigms are highly useful for the quantification of social motivation in mouse models of autism and other disorders of social behavior.

This article describes a new protocol for the quantitative measurement of social motivation in mice using operant conditioning for a social reward. The protocol is useful for the measurement of social motivation in mouse models of autism and other disorders of social behavior.

Quantificare motivazione sociale nei topi mediante condizionamento operante

In this protocol, social motivation is measured in mice through a pair of operant conditioning paradigms. To conduct the experiments, two-chambered ...

Whisker-signaled Eyeblink Classical Conditioning in Head-fixed Mice

Eyeblink conditioning is a common paradigm for investigating the neural mechanisms underlying learning and memory. To better utilize the extensive repertoire of scientific techniques available to study learning and memory at the cellular level, it is ideal to have a stable cranial platform. Because mice do not readily tolerate restraint, they are usually trained while moving about freely in a chamber. Conditioned stimulus (CS) and unconditioned stimulus (US) information are delivered and eyeblink responses recorded via a tether connected to the mouse's head. In the head-fixed apparatus presented here, mice are allowed to run as they desire while their heads are secured to facilitate experimentation. Reliable conditioning of the eyeblink response is obtained with this training apparatus, which allows for the delivery of whisker stimulation as the CS, a periorbital electrical shock as the US, and analysis of electromyographic (EMG) activity from the eyelid to detect blink responses.

The preparation presented here for whisker-signaled eyeblink conditioning in head-fixed mice precisely stimulates specific whiskers while allowing mice to ambulate on a cylindrical treadmill. A whisker stimulation conditioned stimulus (CS) paired with a periorbital shock unconditioned stimulus (US) results in reliable associative learning on this apparatus.

Baffo-segnalato Eyeblink condizionamento classico nei topi Head-fisso

Eyeblink conditioning is a common paradigm for investigating the neural mechanisms underlying learning and memory. To better utilize the extensive ...

Reducing State Anxiety Using Working Memory Maintenance

The purpose of this protocol is to explain how to examine the relationship between working memory processes and anxiety by combining the Sternberg Working Memory (WM) and the threat of shock paradigms. In the Sternberg WM paradigm, subjects are required to maintain a series of letters in the WM for a brief interval and respond by identifying whether the position of a given letter in the series matches a numerical prompt. In the threat of shock paradigm, subjects are exposed to alternating blocks where they are either at risk of receiving unpredictable presentations of a mild electric shock or are safe from the shock. Anxiety is probed throughout the safe and threat blocks using the acoustic startle reflex, which is potentiated under threat (Anxiety-Potentiated Startle (APS)). By conducting the Sternberg WM paradigm during the threat of shock and probing the startle response during either the WM maintenance interval or the intertrial interval, it is possible to determine the effect of WM maintenance on APS.

This protocol demonstrates how to measure anxiety-potentiated startle during the Sternberg Working Memory paradigm.

Ridurre l&#39;ansia di Stato utilizzando la manutenzione della memoria di lavoro

The purpose of this protocol is to explain how to examine the relationship between working memory processes and anxiety by combining the Sternberg Working ...

Classical Short-Delay Eyeblink Conditioning in One-Year-Old Children

Classical eyeblink conditioning (EBC) refers to the learned association between a conditioned stimulus (an auditory tone) and an unconditioned stimulus (a puff of air to the cornea). Eyeblink conditioning is often used experimentally to detect abnormalities in cerebellar-dependent learning and memory that underlies this type of associative learning. While experiments in adults and older children are relatively simple to administer using commercial equipment, eyeblink conditioning in infants is more challenging due to their poor compliance, which makes correct positioning of the equipment difficult. To achieve conditioning in one-year-old infants, a custom-made or an adapted commercial system can be used to deliver the air puff to the infant's cornea. The main challenge lies in successfully detecting and classifying the behavioral responses. We report that automated blink detection methods are unreliable in this population, and that conditioning experiments should be analyzed using frame-by-frame analysis of supplementary video camera recordings. This method can be applied to study developmental changes in eyeblink conditioning and to examine whether this paradigm can detect children with neurological disorders.

This protocol describes an eyeblink conditioning paradigm appropriate for experiments with one-year-old infants. Commercial or custom-made equipment can be used to deliver the stimuli, and data collection and analysis should be performed on the video recordings.

Classica-ritardo Eyeblink condizionata a One-Year-Old Children

Classical eyeblink conditioning (EBC) refers to the learned association between a conditioned stimulus (an auditory tone) and an unconditioned stimulus (a ...

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 ...

Visual Classical Conditioning in Wood Ants

Several species of insects have become model systems for studying learning and memory formation. Although many studies focus on freely moving animals, studies implementing classical conditioning paradigms with harnessed insects have been important for investigating the exact cues that individuals learn and the neural mechanisms underlying learning and memory formation. Here we present a protocol for evoking visual associative learning in wood ants through classical conditioning. In this paradigm, ants are harnessed and presented with a visual cue (a blue cardboard), the conditional stimulus (CS), paired with an appetitive sugar reward, the unconditional stimulus (US). Ants perform a Maxilla-Labium Extension Reflex (MaLER), the unconditional response (UR), which can be used as a readout for learning. Training consists of 10 trials, separated by a 5-minute intertrial interval (ITI). Ants are also tested for memory retention 10 minutes or 1 hour after training. This protocol has the potential to allow researchers to analyze, in a precise and controlled manner, the details of visual memory formation and the neural basis of learning and memory formation in wood ants.

We present a protocol for the classical conditioning of harnessed ants that permits researchers to study visual learning and memory formation at a level of analysis not possible with freely moving individuals.

Visual condizionamento classico in legna formiche

Several species of insects have become model systems for studying learning and memory formation. Although many studies focus on freely moving animals, ...

Fear Incubation Using an Extended Fear-Conditioning Protocol for Rats

Emotional memory has been primarily studied with fear-conditioning paradigms. Fear conditioning is a form of learning through which individuals learn the relationships between aversive events and otherwise neutral stimuli. The most-widely utilized procedures for studying emotional memories entail fear conditioning in rats. In these tasks, the unconditioned stimulus (US) is a footshock presented once or several times across single or several sessions, and the conditioned response (CR) is freezing. In a version of these procedures, called cued fear conditioning, a tone (conditioned stimulus, CS) is paired with footshocks (US) during the training phase. During the first test, animals are exposed to the same context in which training took place, and freezing responses are tested in the absence of footshocks and tones (i.e., a context test). During the second test, freezing is measured when the context is changed (e.g., by manipulating the smell and walls of the experimental chamber) and the tone is presented in the absence of footshocks (i.e., a cue test). Most cued fear conditioning procedures entail few tone-shock pairings (e.g., 1-3 trials in a single session). There is a growing interest in less common versions involving an extensive number of pairings (i.e., overtraining) related to the long-lasting effect called fear incubation (i.e., fear responses increase over time without further exposure to aversive events or conditioned stimuli). Extended fear-conditioning tasks have been key to the understanding of fear incubation&#8217;s behavioral and neurobiological aspects, including its relationship with other psychological phenomena (e.g., post-traumatic stress disorder). Here, we describe an extended fear-conditioning protocol that produces overtraining and fear incubation in rats. This protocol entails a single training session with 25 tone-shock pairings (i.e., overtraining) and a comparison of conditioned freezing responses during context and cue tests 48 h (short-term) and 6 weeks (long-term) after training.

We describe an extended fear-conditioning protocol that produces overtraining and fear incubation in rats. This protocol entails a single training session with 25 tone-shock pairings (i.e., overtraining) and a comparison of conditioned freezing responses during context and cue tests 48 h (short-term) and 6 weeks (long-term) after training.

L'incubazione della paura utilizzando un protocollo esteso di condizionamento della paura per i ratti

Emotional memory has been primarily studied with fear-conditioning paradigms. Fear conditioning is a form of learning through which individuals learn the ...