Questo lavoro è stato sostenuto da una borsa-in-Aid per JSPS Fellows a AA; da sovvenzioni-in-Aid per la Ricerca Scientifica sulle zone innovative per JSP Fellows (n 4501 e n 25.132.712) di TK; e da una borsa di progetto di ricerca assegnato dalla Azabu University. figura 2 è ristampato da Kihara, T., Harada, T., e Koshida, N. Wafer-compatibile fabbricazione e le caratteristiche del silicio nanocristallino emettitori di ultrasuoni indotte termicamente. In: Sensori e Attuatori A: fisiche, volume 125, Elsevier, pag. 426, (2006), con il permesso di Elsevier.

<html> Tutte le procedure sono state approvate dal Comitato Etico di Azabu University. Tutti gli esperimenti sono stati eseguiti in una camera insonorizzata. 1. Preparazione degli animali <ol><li> Maschi per la registrazione <ol><li> Ottenere topi maschi sessualmente maturi esperienza in accoppiamento. </li></ol></li><li> Soggetti di sesso femminile <ol><li> Ottenere vergine topi femmina che hanno ospitato da 2 a 5 fratellini per gabbia (di solito 8 - 12 settimane). </li><li> Ottenere striscio vaginale giornaliero per determinare la fase del ciclo estrale prima della prova, secondo McLean 21. Si deve prestare attenzione a ridurre al minimo la stimolazione vaginale per evitare pseudogravidanza. </li><li> Stain gli strisci con Giemsa, e determinare fase del ciclo estrale in base alla presenza o assenza di leucociti, cellule epiteliali cornified e cellule epiteliali nucleate, secondo Nelson 22. </li></ol></li></ol> 2. Dispositivi (Figura 1) _content &quot;&gt; <img alt="Figura 1" src="/files/ftp_upload/53074/53074fig1.jpg" /> Figura 1. Schema del due scelta Box Test e dispositivi. I topi possono accedere sale A e B attraverso i piccoli cancelli situati tra loro e la zona neutra. La scatola due scelta e due emettitori ultrasuoni sono posti sul pavimento di una camera insonorizzata. Il microfono è sospeso all&#39;interno della camera insonorizzata. Abbreviazione:. PC, personal computer <a href="https://www.jove.com/files/ftp_upload/53074/53074fig1large.jpg" target="_blank">Clicca qui per vedere una versione più grande di questa figura.</a> <ol><li> Per la registrazione <ol><li> Collegare un microfono a condensatore ad ultrasuoni a un PC utilizzando un convertitore USB A / D. </li><li> Impostare il livello di guadagno del convertitore A / D a &quot;7&quot;. </li><li> Dati acustici record in file di formato a 16 bit con una frequenza di campionamento di 400 kHz utilizzando il software di acquisizione audio. </li></ol></li><li> Per la riproduzione <ol><li> Collegare un convertitore D / A al PC utilizzandoun&#39;interfaccia USB. </li><li> Collegare due amplificatori per il convertitore D / A attraverso un attenuatore. (Quest&#39;ultimo dispositivo serve per evitare sovraccarichi.) </li><li> Collegare un emettitore ad ultrasuoni per ogni amplificatore. </li></ol></li></ol> 3. USV registrazione <ol><li> Inserire un topo maschio in piccole (17 x 10 x 11 cm) o medie (21 x 14 x 13 cm) gabbia con un foro di 6 cm di diametro nella parete laterale e il foro coperto con una rete metallica 0,5 cm. Quando si usa una gabbia di medie dimensioni, realizzare una partizione attraverso il centro della gabbia per ridurre l&#39;area. </li><li> Non utilizzare biancheria da letto nella gabbia per evitare fruscii che possono contaminare le registrazioni. </li><li> Collocare un microfono accanto alla mesh. </li><li> Iniziare il monitoraggio ultrasuoni vocalizzazione con l&#39;impostazione di registrazione mostrata al punto 2.1. </li><li> Posizionare un sessualmente maturo topo femmina sia metestrus o diestrus nella gabbia con il topo maschio. </li><li> USV record per 3-5 min. </li></ol> 4. Test Box <ol><li> Costruire la casella prova utilizzando bordo acrilico (Figura 1). Nota: La casella di prova (alta 35 x 20, e x 20 cm) è diviso in tre scomparti: una zona neutra (15 x 20 cm), e sale A e B (20 x 10 cm, ciascuno). Camere A e B hanno ciascuno un foro di 4 centimetri di diametro che è coperto con una rete metallica 0,5 centimetri. I fori e tele si trovano a ciascuna estremità del sale A e B. Fai cancelli (5 x 5 cm ciascuno) tra la zona neutra e camere A e B. </li><li> Per evitare perdite suono isolare ciascun emettitore ultrasuoni, ad eccezione del lato rivolto verso la rete, utilizzando lastre in gomma, loro tenuta alla scatola intorno al perimetro di ciascuna maglia. </li><li> Sigilli di piombo posto su entrambi i lati del muro tra le stanze A e B per l&#39;isolamento acustico. </li></ol> 5. La preparazione per la riproduzione del suono <ol><li> Creazione di riproduzione Suoni <ol><li> Catturare un segmento di 20 secondi di USV dal file registrato utilizzando software di editing audio digitale. </li><li> Selezionare e catturare sfondo noise (0.35 sec) dal file segmento 20 sec USV. Creare un file di rumore di fondo 20 sec ripetendo il segmento rumore. </li><li> Per riprodurre i due file audio contemporaneamente, esportarli come un unico file audio stereo, con un file per il &quot;orecchio sinistro&quot; e l&#39;altro file per il &quot;orecchio destro.&quot; </li><li> Filtrare i file audio utilizzando come filtro passa-alto con un taglio di 40 kHz. </li><li> Ridurre il rumore utilizzando lo strumento di riduzione del rumore in software di post-elaborazione. Selezionare e catturare un segmento del file che contiene USV come il profilo di riduzione del rumore, ed eseguire la riduzione del rumore con il livello 40. </li></ol></li><li> Taratura di riproduzione Suoni <ol><li> Assicurarsi che lo spazio tra gli emettitori di ultrasuoni e il microfono è di 10 cm. </li><li> Per calibrare i livelli di pressione sonora dagli emettitori di ultrasuoni, effettuare il monitoraggio ad ultrasuoni utilizzando la configurazione descritta al punto 2.1. Misurare il livello di pressione sonora dal microfono in decibel (dB). </li><li> Using l&#39;attenuatore e amplificatori, regolare il volume dei USV generati dai emettitori ultrasuoni allo stesso livello di pressione sonora come USV maschi registrate nel passaggio 3. </li><li> Quando si utilizzano i due emettitori di ultrasuoni e un file audio composto da USV e rumore di fondo, verificare che i due emettitori di ultrasuoni presentano lo stesso livello di pressione sonora con un suono di calibrazione (per esempio, un tono puro 75-kHz) prima di eseguire il test comportamentale. </li><li> Quando si riproduce un file audio composto da due flussi di USV, regolare i due livelli di pressione sonora generati allo stesso livello, prima del test. </li></ol></li></ol> 6. Prova a due scelta <ol><li> Posizionare gli emettitori di ultrasuoni dietro le maglie della scatola della prova. </li><li> Chiudere le porte di stanze A e B con un bordo acrilico, e abituare il soggetto femminile alla zona neutra per 30 min. Coprire la scatola con bordo acrilico per evitare che il mouse può sfuggire. </li><li> Avviare la registrazione video utilizzando un CCFotocamera D montata sopra la casella. La telecamera (non mostrato in figura 1) comprende una superficie di tutte tre scomparti. </li><li> Rimuovere il cancello e la copertura di bordo e lasciare la femmina di esplorare liberamente la casella di prova, comprese le zone sane. </li><li> Una volta che la femmina ha indagato due maglie, ed è tornato alla zona neutra, avviare la riproduzione di un file audio di 20 secondi più volte per 5 o 10 minuti. </li><li> Condurre il monitoraggio comportamentale per 5 o 10 minuti. </li><li> Per ridurre stimoli olfattivi indesiderati depositati dal soggetto precedente, pulire le gabbie di prova tra le prove con il 70% di etanolo. </li><li> Accendere il luogo di emettitori di ultrasuoni A e B in modo casuale al fine di evitare pregiudizi inerenti effetto lato verso le camere A o B. </li></ol> 7. Analisi statistica <ol><li> Utilizzando il software comportamentale evento-gol, l&#39;analisi dei seguenti parametri: numero totale delle voci in ogni stanza; durata totale di permanenza in ogni zona mediana; durata totale di permanenza in ogni zona del suono; edurata totale di ricerca di ciascuna maglia davanti ultrasuoni emettitore. Figura 1 illustra ciò che si intende per &quot;zona intermedia&quot; e &quot;zona suono.&quot; Nota: Nei casi in cui il mouse rimane sulla linea di separazione tra le due zone (figura 1, linea tratteggiata) decidere la posizione della direzione della testa. Quando la testa rivolta verso il lato della maglia, è segnato come permanenza nella zona di suono. </li><li> Per ogni parametro comportamentale, confrontare le risposte per camere A e B con un Wilcoxon prova firmato-rank, o t -test in coppia con un livello di significatività di 0.05. </li></ol>

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The authors declare that they have no competing interests.

Here, the results of a representative test showed that female mice can discriminate between artificial male USVs and background noise (Figure 3). The conclusion to be drawn from these results is that the discrimination signal is reflected in the duration of stay in the room and sound zone, and in the duration of searching the mesh in the first 5 min of testing, but not in the second 5 min (Figure 3C, E and F). These data indicate that mice become habituated to the playback sounds, possib...

Molti animali usano vocalizzazioni per la comunicazione intraspecifica. Nel topo Mus musculus, uno importante tipo di segnale di comunicazione è vocalizzazioni ultrasoniche (USV), che hanno frequenze superiori a 20 kHz. USV emessi dai topi sono considerati una componente di riconoscimento sociale in maschio-femmina 1-4, femmina-femmina 1, 5, e maschio-maschio 1, 6 interazioni. USV vengono emessi anche da cuccioli quando sono isolate dalla loro madre, che aumenta il suo comportamento-pup recupero, e di conseguenza la sopravvivenza Pup 7. Anche se molti studi hanno analizzato e classificato USV del mouse 8, 9, le risposte comportamentali e meccanismi neurali dell&#39;animale ricevente sono stati meno documentati 10, 11. Quest&#39;ultimo è necessario per chiarire il significato biologico delle diverse caratteristiche di USV. Per rivelare questi meccanismi, l&#39;esperimento di riproduzione è un metodo efficiente. Studi di riproduzione recenti hanno rivelato che femminilei topi sono attratti da USV 12, e che preferiscono USV da maschi che sono diversi dai loro genitori 13, 14. Questo articolo spiega il test di riproduzione utilizzato per valutare USV preferenza nei topi. Una scatola test a due scelta è stata sviluppata in cui due USV differenti possono essere riprodotte contemporaneamente in due compartimenti di un contenitore di prova, come illustrato in Figura 1. Questo tipo di scatola di prova impedisce la contaminazione suono dividendo l&#39;area di prova in tre sottocamere , con muri di piombo. Gli emettitori di ultrasuoni si trovano al di fuori di ogni stanza. Nel muro tra le camere e emettitori di ultrasuoni sono buchi coperti di rete metallica. Topi può muoversi liberamente nelle tre sale, e mostra una &quot;ricerca della rete&quot; comportamento, come se per rispondere a USV riprodotte dagli emettitori di ultrasuoni. In questo test, i topi soggiornare per periodi di durata diversa vicino a un emettitore sonoro o l&#39;altro. Questi parametri possono essere registrati per ottenere un sensibile misu<html> re di preferenza suono. Per riprodurre i USV indietro, silicio nanocristallino emettitori termo-acustico (cioè, &quot;emettitore nc-Si&quot;), sono stati utilizzati come in studi precedenti 15-17. Questi dispositivi sono composti da un elettrodo a film sottile riscaldatore, uno strato di silicio poroso nano, e un wafer di silicio monocristallino. Il file audio digitale viene convertito in un segnale analogico e poi fatto passare attraverso l&#39;elettrodo riscaldatore. Il dispositivo converte i segnali termici derivanti voltaggio-dipendenti in significativo di pressione sonora con bassa distorsione. Questo dispositivo è unico in quanto, a differenza di generatori di suoni comuni che dipendono da vibrazioni meccaniche, può riprodurre il suono senza la necessità di un diaframma. L&#39;emettitore presenta un livello di pressione sonora piatta a frequenze da 20 a 160 kHz (figura 2), e può riprodurre USV murini digitalmente registrate molto accuratamente in termini di durata, frequenza e livello sonoro di pressione 15, 18, ​​19. ve_content &quot;&gt; In un esperimento rappresentativo mostrato in Figura 3, C57BL / 6 (B6) femmine hanno permesso di scegliere tra BALB / c (BALB) USV maschi e rumore di fondo. Inoltre, la Figura 4 mostra la scelta delle femmine B6 e BALB tra riproduzioni USV simultanee da un topo BALB e un maschio B6, come riportato in un precedente studio 14. Le caratteristiche di USV maschili differiscono B6 e BALB ceppi 20. Come mostrato da questi risultati, l&#39;attrattiva di USV può essere valutata con il presente protocollo, in cui i suoni sono registrati da un individuo dal vivo, acusticamente analizzati e riprodotti ad altri individui.

<table><tbody><tr><td>Soundproof chamber</td><td>Muromachi Kikai</td><td></td><td></td></tr><tr><td>Small cage</td><td>CLEA Japan</td><td>CL-0113-1</td><td></td></tr><tr><td>Middle cage</td><td>CLEA Japan</td><td>CL-0103-1</td><td></td></tr><tr><td>Ultrasound condenser microphones</td><td>Avisoft Bioacoustics</td><td>CM16/CMPA</td><td></td></tr><tr><td>A/D converter</td><td>Avisoft Bioacoustics</td><td>UltraSoundGate116H</td><td></td></tr><tr><td>Audio software</td><td>Avisoft Bioacoustics</td><td>RECORDER USGH</td><td></td></tr><tr><td>Adobe Audition 3.0 / Audio editing software</td><td>Adobe Systems</td><td>Adobe Audition 3.0</td><td></td></tr><tr><td>Nc-Si emitter</td><td>Original</td><td></td><td>not commercially available but it is planned to be so in near future</td></tr><tr><td>D/A converter</td><td>National Instruments</td><td>NI USB-6251 BNC</td><td></td></tr><tr><td>Attenuator</td><td>Original</td><td></td><td></td></tr><tr><td>Amplifier</td><td>Yamatake</td><td></td><td></td></tr><tr><td>PC</td><td>Windows 7 professional</td><td></td><td>Intel&amp;reg; core i7-2600K CPU @ 3.4GHz, 8GB RAM, 64-bit operating system</td></tr><tr><td>Event recorder Excel-macro / Event-scoring software</td><td>original</td><td></td><td>Programmed by Naoto Akagawa &amp;amp; Takeru Yamamoto</td></tr><tr><td>CCD Camera</td><td></td><td></td><td></td></tr><tr><td>Rubber plates (made of elastomer resin)</td><td>Tokyo bouon</td><td>TI-75BK B4</td><td>Cut them to the proper size http://www.piano-bouon.jp/shopping/?pid=1329272401-447630&amp;amp;ca=6&amp;amp;p=3</td></tr><tr><td>Giemsa&#039;s azur eosin methylene blue solution</td><td>Merck Millipore</td><td>1.09204.0500</td><td></td></tr></tbody></table>

determining ultrasonic vocalization preferences in mice using a two-choice playback test

Mice emit ultrasonic vocalizations (USVs) during a variety of conditions, such as pup isolation and adult social interactions. These USVs differ with age, sex, condition, and genetic background of the emitting animal. Although many studies have characterized these differences, whether receiver mice can discriminate among objectively different USVs and show preferences for particular sound traits remains to be elucidated. To determine whether mice can discriminate between different characteristics of USVs, a playback experiment was developed recently, in which preference responses of mice to two different USVs could be evaluated in the form of a place preference.
First, USVs from mice were recorded. Then, the recorded USVs were edited, trimmed accordingly, and exported as stereophonic sound files. Next, the USV amplitudes generated by the two ultrasound emitters used in the experiment were adjusted to the same sound pressure level. Nanocrystalline silicon thermo-acoustic emitters were used to play the USVs back. Finally, to investigate the preference of subject mice to selected USVs, pairs of two differing USV signals were played back simultaneously in a two-choice test box. By repeatedly entering a defined zone near an ultrasound emitter and searching the wire mesh in front of the emitter, the mouse reveals its preference for one sound over another. This model allows comparing the attractiveness of the various features of mouse USVs, in various contexts.

I USV registrate da un topo BALB-maschio (161 sillabe per 20 sec), così come rumore di fondo sono stati utilizzati come riprodurre suoni nell&#39;esperimento rappresentante illustrato nella figura 3. In questo esperimento, 7 topi B6 femminili sono stati usati a 9 settimane di età . Per determinare la migliore durata della prova della risposta femminile di riproduzione suoni, i parametri comportamentali sono state analizzate separatamente per i primi e gli ultimi cinque minuti del tempo di collaudo tot...

Watch this Scientific Journal Video about Determining Ultrasonic Vocalization Preferences in Mice using a Two-choice Playback Test at JoVE.com

Determining Ultrasonic Vocalization Preferences in Mice using a Two-choice Playback Test

Ultrasonic vocalizations (USVs) in mice differ depending on age, sex, condition, and genetic background. Using two ultrasound emitters broadcasting simultaneously in different locations, this two-choice test can evaluate murine recognition and preference responses to different characteristics of USVs.

Determinazione Preferenze ultrasuoni vocalizzazione nei topi utilizzando un test di riproduzione a due scelta

Mice emit ultrasonic vocalizations (USVs) during a variety of conditions, such as pup isolation and adult social interactions. These USVs differ with age, ...

determining-ultrasonic-vocalization-preferences-mice-using-two-choice

Research

JoVE Journal

Behavior

11.1K Views.  Azabu University. Ultrasonic vocalizations (USVs) in mice differ depending on age, sex, condition, and genetic background. Using two ultrasound emitters broadcasting simultaneously in different locations, this two-choice test can evaluate murine recognition and preference responses to different characteristics of USVs.

Video: Determining Ultrasonic Vocalization Preferences in Mice using a Two-choice Playback Test

Assessment of Ultrasonic Vocalizations During Drug Self-administration in Rats

Drug self-administration procedures are commonly used to study behavioral and neurochemical changes associated with human drug abuse, addiction and relapse. Various types of behavioral activity are commonly utilized as measures of drug motivation in animals. However, a crucial component of drug abuse relapse in abstinent cocaine users is "drug craving", which is difficult to model in animals, as it often occurs in the absence of overt behaviors. Yet, it is possible that a class of ultrasonic vocalizations (USVs) in rats may be a useful marker for affective responses to drug administration, drug anticipation and even drug craving. Rats vocalize in ultrasonic frequencies that serve as a communicatory function and express subjective emotional states. Several studies have shown that different call frequency ranges are associated with negative and positive emotional states. For instance, high frequency calls ("50-kHz") are associated with positive affect, whereas low frequency calls ("22-kHz") represent a negative emotional state. This article describes a procedure to assess rat USVs associated with daily cocaine self-administration. For this procedure, we utilized standard single-lever operant chambers housed within sound-attenuating boxes for cocaine self-administration sessions and utilized ultrasonic microphones, multi-channel recording hardware and specialized software programs to detect and analyze USVs. USVs measurements reflect emotionality of rats before, during and after drug availability and can be correlated with commonly assessed drug self-administration behavioral data such lever responses, inter-response intervals and locomotor activity. Since USVs can be assessed during intervals prior to drug availability (e.g., anticipatory USVs) and during drug extinction trials, changes in affect associated with drug anticipation and drug abstinence can also be determined. In addition, determining USV changes over the course of short- and long-term drug exposure can provide a more detailed interpretation of drug exposure effects on affective functioning.

Drug self-administration and ultrasonic vocalizations (USV) are used as behavioral assessments in animal research, but rarely in combination. The purpose of this article is to describe the advantages of recording USVs during drug self-administration procedures to assess affective responses to drug experience.

Valutazione di vocalizzazioni ultrasoniche Durante Drug autosomministrazione in topi

Drug self-administration procedures are commonly used to study behavioral and neurochemical changes associated with human drug abuse, addiction and ...

Ricerca

Neuroscienze

Recording Mouse Ultrasonic Vocalizations to Evaluate Social Communication

Mice emit ultrasonic vocalizations in different contexts throughout development and in adulthood. These vocal signals are now currently used as proxies for modeling the genetic bases of vocal communication deficits. Characterizing the vocal behavior of mouse models carrying mutations in genes associated with neuropsychiatric disorders such as autism spectrum disorders will help to understand the mechanisms leading to social communication deficits. We provide here protocols to reliably elicit ultrasonic vocalizations in pups and in adult mice. This standardization will help reduce inter-study variability due to the experimental settings. Pup isolation calls are recorded throughout development from individual pups isolated from dam and littermates. In adulthood, vocalizations are recorded during same-sex interactions (without a sexual component) by exposing socially motivated males or females to an unknown same-sex conspecific. We also provide a protocol to record vocalizations from adult males exposed to an estrus female. In this context, there is a sexual component in the interaction. These protocols are established to elicit a large amount of ultrasonic vocalizations in laboratory mice. However, we point out the important inter-individual variability in the vocal behavior of mice, which should be taken into account by recording a minimal number of individuals (at least 12 in each condition). These recordings of ultrasonic vocalizations are used to evaluate the call rate, the vocal repertoire and the acoustic structure of the calls. Data are combined with the analysis of synchronous video recordings to provide a more complete view on social communication in mice. These protocols are used to characterize the vocal communication deficits in mice lacking ProSAP1/Shank2, a gene associated with autism spectrum disorders. More ultrasonic vocalizations recordings can also be found on the mouseTube database, developed to favor the exchange of such data.

Mouse ultrasonic vocalizations are used as proxies to model the genetic bases of vocal communication deficits in mouse models for neuropsychiatric disorders. The present protocol describes three experimental contexts that reliably elicit ultrasonic vocalizations from pups (throughout development) and adult mice (same-sex interactions, male-estrus female interactions).

Registrazione del mouse ultrasuoni Vocalizations per valutare Comunicazione Sociale

Mice emit ultrasonic vocalizations in different contexts throughout development and in adulthood. These vocal signals are now currently used as proxies ...

Comportamento

Targeted Training of Ultrasonic Vocalizations in Aged and Parkinsonian Rats

Voice deficits are a common complication of both Parkinson disease (PD) and aging; they can significantly diminish quality of life by impacting communication abilities. 1, 2 Targeted training (speech/voice therapy) can improve specific voice deficits,3, 4 although the underlying mechanisms of behavioral interventions are not well understood. Systematic investigation of voice deficits and therapy should consider many factors that are difficult to control in humans, such as age, home environment, age post-onset of disease, severity of disease, and medications. The method presented here uses an animal model of vocalization that allows for systematic study of how underlying sensorimotor mechanisms change with targeted voice training. The ultrasonic recording and analysis procedures outlined in this protocol are applicable to any investigation of rodent ultrasonic vocalizations.
The ultrasonic vocalizations of rodents are emerging as a valuable model to investigate the neural substrates of behavior.5-8 Both rodent and human vocalizations carry semiotic value and are produced by modifying an egressive airflow with a laryngeal constriction.9, 10 Thus, rodent vocalizations may be a useful model to study voice deficits in a sensorimotor context. Further, rat models allow us to study the neurobiological underpinnings of recovery from deficits with targeted training.
To model PD we use Long-Evans rats (Charles River Laboratories International, Inc.) and induce parkinsonism by a unilateral infusion of 7 &mu;g of 6-hydroxydopamine (6-OHDA) into the medial forebrain bundle which causes moderate to severe degeneration of presynaptic striatal neurons (for details see Ciucci, 2010).11, 12 For our aging model we use the Fischer 344/Brown Norway F1 (National Institute on Aging).
Our primary method for eliciting vocalizations is to expose sexually-experienced male rats to sexually receptive female rats. When the male becomes interested in the female, the female is removed and the male continues to vocalize. By rewarding complex vocalizations with food or water, both the number of complex vocalizations and the rate of vocalizations can be increased (Figure 1).
An ultrasonic microphone mounted above the male's home cage records the vocalizations. Recording begins after the female rat is removed to isolate the male calls. Vocalizations can be viewed in real time for training or recorded and analyzed offline. By recording and acoustically analyzing vocalizations before and after vocal training, the effects of disease and restoration of normal function with training can be assessed. This model also allows us to relate the observed behavioral (vocal) improvements to changes in the brain and neuromuscular system.

Voice disorders are debilitating in aging and Parkinson disease. The ultrasonic vocalizations of rats, also affected by these conditions, can be used to study these voice disorders, their neural substrates, and the nature of functional recovery with behavioral intervention.

Formazione mirata delle vocalizzazioni ultrasoniche in ratti anziani e Parkinson

Voice deficits are a common complication of both Parkinson disease (PD) and aging; they can significantly diminish quality of life by impacting ...

Eliciting and Analyzing Male Mouse Ultrasonic Vocalization (USV) Songs

Mice produce ultrasonic vocalizations (USVs) in a variety of social contexts throughout development and adulthood. These USVs are used for mother-pup retrieval1, juvenile interactions2, opposite and same sex interactions3,4,5, and territorial interactions6. For decades, the USVs have been used by investigators as proxies to study neuropsychiatric and developmental or behavioral disorders7,8,9, and more recently to understand mechanisms and evolution of vocal communication among vertebrates10. Within the sexual interactions, adult male mice produce USV songs, which have some features similar to courtship songs of songbirds11. The use of such multisyllabic repertoires can increase potential flexibility and information they carry, as they can be varied in how elements are organized and recombined, namely syntax. In this protocol a reliable method to elicit USV songs from male mice in various social contexts, such as exposure to fresh female urine, anesthetized animals, and estrus females is described. This includes conditions to induce a large amount of syllables from the mice. We reduce recording of ambient noises with inexpensive sound chambers, and present a quantification method to automatically detect, classify and analyze the USVs. The latter includes evaluation of call-rate, vocal repertoire, acoustic parameters, and syntax. Various approaches and insight on using playbacks to study an animal&#39;s preference for specific song types are described. These methods were used to describe acoustic and syntax changes across different contexts in male mice, and song preferences in female mice.

Eliciting and Analyzing Male Mouse Ultrasonic Vocalization USV Songs

Mice produce a complex multisyllabic repertoire of ultrasonic vocalizations (USVs). These USVs are widely used as readouts for neuropsychiatric disorders. This protocol describes some of the practices we learned and developed to consistently induce, collect, and analyze the acoustic features and syntax of mouse songs.

Eliciting e analisi di canzoni ad ultrasuoni (USV) di topo maschio

Mice produce ultrasonic vocalizations (USVs) in a variety of social contexts throughout development and adulthood. These USVs are used for mother-pup ...

Introducing Clicker Training as a Cognitive Enrichment for Laboratory Mice

Establishing new refinement strategies in laboratory animal science is a central goal in fulfilling the requirements of Directive 2010/63/EU. Previous research determined a profound impact of gentle handling protocols on the well-being of laboratory mice. By introducing clicker training to the keeping of mice, not only do we promote the amicable treatment of mice, but we also enable them to experience cognitive enrichment. Clicker training is a form of positive reinforcement training using a conditioned secondary reinforcer, the "click" sound of a clicker, which serves as a time bridge between the strengthened behavior and an upcoming reward. The effective implementation of the clicker training protocol with a cohort of 12 BALB/c inbred mice of each sex proved to be uncomplicated. The mice learned rather quickly when challenged with tasks of the clicker training protocol, and almost all trained mice overcame the challenges they were given (100% of female mice and 83% of male mice). This study has identified that clicker training for mice strongly correlates with reduced fear in the mice during human-mice interactions, as shown by reduced anxiety-related behaviors (e.g., defecation, vocalization, and urination) and fewer depression-like behaviors (e.g., floating). By developing a reliable protocol that can be easily integrated into the daily routine of the keeping of laboratory mice, the lifetime experience of welfare in the mice can be improved substantially.

The development of new refinement strategies for laboratory mice is a challenging task that contributes towards fulfilling the 3R principle. This protocol introduces clicker training as a cognitive enrichment program for laboratory mice.

L&#39;introduzione di Clicker Training come un arricchimento conoscitivo per topi di laboratorio

Establishing new refinement strategies in laboratory animal science is a central goal in fulfilling the requirements of Directive 2010/63/EU. Previous ...

A Behavioral Assay for Investigating the Role of Spatial Memory During Instinctive Defense in Mice

Evolution has selected a repertoire of defensive behaviors that are essential for survival across all animal species. These behaviors are often stereotyped actions elicited in response to innately aversive sensory stimuli, but their success requires enough flexibility for adapting to different spatial environments, which can change rapidly. Here, we describe a behavioral assay to evaluate the influence of learned spatial knowledge on defensive behaviors in mice. We have adapted the widely used Barnes maze spatial memory assay to investigate how mice navigate to a shelter during escape responses to innately aversive sensory stimuli in a novel environment, and how they adapt to acute changes in the environment. This new assay is an ethological paradigm that does not require training and exploits the natural exploration patterns and navigation strategies in mice. We propose that the set of protocols described here are a powerful means of studying goal-directed behaviors and stimulus-triggered navigation, which should be of interest to both the fields of instinctive behaviors and spatial memory.

This protocol describes a behavioral assay, based on the Barnes maze test, to study how instinctive defensive actions are modified by the knowledge of spatial environment.

Un'analisi comportamentale per investigare il ruolo della memoria spaziale durante la difesa istintiva nei topi

Evolution has selected a repertoire of defensive behaviors that are essential for survival across all animal species. These behaviors are often ...

A Low Cost Setup for Behavioral Audiometry in Rodents

In auditory animal research it is crucial to have precise information about basic hearing parameters of the animal subjects that are involved in the experiments. Such parameters may be physiological response characteristics of the auditory pathway, e.g. via brainstem audiometry (BERA). But these methods allow only indirect and uncertain extrapolations about the auditory percept that corresponds to these physiological parameters. To assess the perceptual level of hearing, behavioral methods have to be used. A potential problem with the use of behavioral methods for the description of perception in animal models is the fact that most of these methods involve some kind of learning paradigm before the subjects can be behaviorally tested, e.g. animals may have to learn to press a lever in response to a sound. As these learning paradigms change perception itself 1,2 they consequently will influence any result about perception obtained with these methods and therefore have to be interpreted with caution. Exceptions are paradigms that make use of reflex responses, because here no learning paradigms have to be carried out prior to perceptual testing. One such reflex response is the acoustic startle response (ASR) that can highly reproducibly be elicited with unexpected loud sounds in na&iuml;ve animals. This ASR in turn can be influenced by preceding sounds depending on the perceptibility of this preceding stimulus: Sounds well above hearing threshold will completely inhibit the amplitude of the ASR; sounds close to threshold will only slightly inhibit the ASR. This phenomenon is called pre-pulse inhibition (PPI) 3,4, and the amount of PPI on the ASR gradually depends on the perceptibility of the pre-pulse. PPI of the ASR is therefore well suited to determine behavioral audiograms in na&iuml;ve, non-trained animals, to determine hearing impairments or even to detect possible subjective tinnitus percepts in these animals. In this paper we demonstrate the use of this method in a rodent model (cf. also ref. 5), the Mongolian gerbil (Meriones unguiculatus), which is a well know model species for startle response research within the normal human hearing range (e.g. 6).

A fast and inexpensive method for the behavioral determination of hearing parameters like hearing thresholds, hearing impairments or phantom perceptions (subjective tinnitus) is described. It uses pre-pulse inhibition of the acoustic startle response and can be easily implemented in a personal computer using a programmable AD/DA-converter and a piezo sensor.

Un programma di installazione a basso costo per audiometria comportamentale nei roditori

In auditory animal research it is crucial to have precise information about basic hearing parameters of the animal subjects that are involved in the ...

A Novel Pavlovian Fear Conditioning Paradigm to Study Freezing and Flight Behavior

Fear- and anxiety-related behaviors significantly contribute to an organism&#8217;s survival. However, exaggerated defensive responses to perceived threat are characteristic of various anxiety disorders, which are the most prevalent form of mental illness in the United States. Discovering the neurobiological mechanisms responsible for defensive behaviors will aid in the development of novel therapeutic interventions. Pavlovian fear conditioning is a widely used laboratory paradigm to study fear-related learning and memory. A major limitation of traditional Pavlovian fear conditioning paradigms is that freezing is the only defensive behavior monitored. We recently developed a modified Pavlovian fear conditioning paradigm that allows us to study both conditioned freezing and flight (also known as escape) behavior within individual subjects. This model employs higher intensity footshocks and a greater number of pairings between the conditioned stimulus and unconditioned stimulus. Additionally, this conditioned flight paradigm utilizes serial presentation of pure tone and white noise auditory stimuli as the conditioned stimulus. Following conditioning in this paradigm, mice exhibit freezing behavior in response to the tone stimulus, and flight responses during the white noise. This conditioning model can be applied to the study of rapid and flexible transitions between behavioral responses necessary for survival.

Defensive behavioral responses are contingent upon threat intensity, proximity, and context of exposure. Based on these factors, we developed a classical conditioning paradigm that elicits clear transitions between conditioned freezing and flight behavior within individual subjects. This model is crucial for the understanding the pathologies involved in anxiety, panic, and post-traumatic stress disorders.

Un nuovo paradigma pavloviano di condizionamento della paura per studiare il congelamento e il comportamento di volo

Fear- and anxiety-related behaviors significantly contribute to an organism&#8217;s survival. However, exaggerated defensive responses to perceived threat ...

Studio dei comportamenti difensivi attivi nei topi con condizionamento della paura modificato

Modified Fear Conditioning for Inducing Flight Behaviors in Mice

The appropriate manifestation of defensive behavior in a threatening situation is critical for survival. The prevailing theory suggests that an active defensive behavior, such as jumping or rapid darting, is expressed under high threat imminence or actual threat, whereas passive defensive behavior, such as freezing, is expressed when the threat is predicted, but the threat imminence is relatively low. In classical fear conditioning, subjects typically exhibit freezing as a conditioned defensive response, with little expression of active defensive behavior in most cases. Here, we introduce a modified fear conditioning procedure for mice to observe the transition from freezing to flight and vice versa, involving five repetitive pairings of conditioned stimuli (CS; continuous tone, 8 kHz, 95 dB SPL (sound pressure levels)) and unconditioned stimuli (US; foot shock, 0.9 mA, 1.0 s) over two days. This modified fear conditioning procedure requires a relatively large number of conditioning sessions and conditioning days but does not necessitate a high-intensity foot shock for modest expression of flight behavior. Using the same context for conditioning and salient CS presentations is essential to elicit flight behaviors. This modified fear conditioning procedure is a reliable method for observing active defensive behaviors in mice, providing an opportunity to elucidate the fine mechanisms and characteristics of such behaviors in a fearful context.

Autore in primo piano: Esplorazione dei correlati neurali dei comportamenti difensivi nell'apprendimento e nell'estinzione della paura

To study flight behaviors in a fearful context, we introduce a modified fear conditioning protocol. This protocol ensures that mice consistently exhibit flight behaviors during cue presentation in the fear conditioning.

Condizionamento della paura modificato per indurre comportamenti di volo nei topi

The appropriate manifestation of defensive behavior in a threatening situation is critical for survival. The prevailing theory suggests that an active ...

Determinazione Preferenze ultrasuoni vocalizzazione nei topi utilizzando un test di riproduzione a due scelta

Riepilogo

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Capitoli in questo video

Valutazione di vocalizzazioni ultrasoniche Durante Drug autosomministrazione in topi

Registrazione del mouse ultrasuoni Vocalizations per valutare Comunicazione Sociale

Formazione mirata delle vocalizzazioni ultrasoniche in ratti anziani e Parkinson

Eliciting e analisi di canzoni ad ultrasuoni (USV) di topo maschio

L'introduzione di Clicker Training come un arricchimento conoscitivo per topi di laboratorio

Un'analisi comportamentale per investigare il ruolo della memoria spaziale durante la difesa istintiva nei topi

Un programma di installazione a basso costo per audiometria comportamentale nei roditori

Un nuovo paradigma pavloviano di condizionamento della paura per studiare il congelamento e il comportamento di volo

Condizionamento della paura modificato per indurre comportamenti di volo nei topi

Condizionamento della paura modificato per indurre comportamenti di volo nei topi