We thank Rachelle Kanippayoor for her help in identifying the new wild-type strain as being of the melanogaster species. R.W.F, O.F. and A.F.S were responsible for research design; R.W.F, M.N. and O.F. performed the experiments. R.W.F, M.N., O.F. and A.F.S. analyzed the data; R.W.F., I.S.M. and A.F.S. wrote the manuscript. 
This work was supported by PSC-CUNY research awards, jointly funded by The Professional Staff Congress and The City University of New York to A.F.S.; by internal funding from Western University to A.F.S. and I.S.M.; by a training support from the National Alliance for Hispanic Health&#8217;s Alliance/Merck Ciencia (Science) Hispanic Scholars Program and a University Fellowship from the Yale Graduate School of Arts and Sciences to R.W.F.

1. Attrezzature e reagenti creati in-house (vedi elenco dei materiali per gli altri) <ol><li> Preparare una Drosophila apparecchiatura anestesia fredda per svolgere un lavoro mosca. <ol><li> Tagliare un foglio di polietilene porosa per coprire una piccola scatola di plastica (lungo 12,7 centimetri, larghezza 10,2 centimetri), in genere la copertura superiore di una scatola punte per pipette (lungo 12,7 centimetri, larghezza 10,2 centimetri, 3,8 centimetri di profondità). </li><li> Riempire la scatola con ghiaccio tritato, coprire con il foglio di polietilene porosa. </li></ol></li><li> Preparare un apparato T-labirinto; adattare l&#39;apparecchiatura precedentemente descritta in dettaglio 28-31. </li><li> Preparare un aspiratore mosca come descritto 32. </li><li> Preparare un apparato di risposta fototassi per spostare le mosche dentro e fuori di flaconi di prova, senza stress: <ol><li> Utilizzare un apparato in controcorrente, come descritto da 29,33. </li></ol></li></ol> NOTA: La T-maze può essere utilizzato anche per altri scopi, come descritto da 28. <ol start="5"><li> Garantire homcondizioni di luce ogeneous: eseguire l&#39;esperimento su un banco coperto con un coperchio panchina bianca, e di fronte a un bordo bianco. NOTA: per questi esperimenti, i ceppi utilizzati sono stati mosca Drosophila melanogaster wild-tipo: Canton-S, Oregon e Samarcanda vola dalle nostre scorte di laboratorio 34; Mosche Elwood sono stati raccolti in autunno 2011 nel quartiere Elwood di Huntington, a Long Island, New York, Stati Uniti d&#39;America. </li></ol> 2. Preparare le mosche prima dell&#39;esperimento 1-2 giorni prima di eseguire l&#39;esperimento: <ol><li> Mantenere mosche sul medio farina di mais-agar-melassa-lievito di serie a 25 ° C su un 12 ore di luce / buio ciclo. </li><li> Raccogliere linea risponditore sotto anestesia freddi 1-2 giorni prima di eseguire l&#39;esperimento <ol><li> Trasferire le mosche dalle bottiglie in cui sono stati raccolti in 50 ml provette Falcon, con un imbuto. </li><li> Posizionare il tubo falco con le mosche sotto il livello del ghiaccio, in unsecchiello per il ghiaccio isolato. </li><li> Collocare l&#39;apparecchio di anestesia fredda con ghiaccio tritato a -4 ° C per rilassarsi. </li><li> Attendere 5 minuti per le mosche di entrare in un coma freddo. </li><li> Trasferire le mosche sul foglio di polietilene porosa dell&#39;apparato anestesia freddo. </li><li> Raccogliere le mosche risponditore sull&#39;apparato anestesia freddo. A RT, sotto uno stereomicroscopio, separare i 3-7 giorni vecchio maschio da mosche femmine e mantenere queste mosche in gruppi di 40. </li></ol></li><li> Raccogliere linea emettitore 1-2 giorni prima di eseguire l&#39;esperimento. <ol><li> Raccoglie delicatamente Canton-S mixed-sex aria dal loro bottiglie, con un aspiratore bocca, controllando per il loro numero. </li><li> Mantenere queste mosche in gruppo di 60-100, in quanto saranno gli emettitori. </li><li> Preparare tutti i campioni di emettitori come punti dati necessari. </li></ol></li><li> Lasciare almeno una notte di recupero dopo la raccolta dei soccorritori e per gli emettitori, per ridurre al minimo qualsiasi effetto confondente dell&#39;anestesia freddoe la raccolta della bocca di aspirazione sul comportamento. </li><li> Aprire la busta di fiale di prova, per permettere all&#39;aria di plastica maleodoranti stantio nel sacchetto per essere sostituito da aria fresca, almeno 1 giorno prima dell&#39;esperimento. </li></ol> 2 ore prima di eseguire l&#39;esperimento: <ol start="6"><li> Assicurarsi che la temperatura nel locale in cui viene eseguita l&#39;esperimento è di circa 23-25 ​​° C, con anche la luce, e umidità superiore al 30%. </li><li> Trasferire i soccorritori e mosche emettitori di frutta in fiale di cibo fresco per garantire che queste mosche non sono affamate. </li><li> Lasciate che i moscerini della frutta regolare per l&#39;ambiente per 2 ore, sulla panchina su cui verrà eseguita l&#39;esperimento. </li></ol> 3. Eseguire l&#39;esperimento sociale Prevenzione <ol><li> Eseguire ogni esperimento allo stesso tempo del giorno, in un intervallo di 3-4 ore nel pomeriggio tra tempo Zeitgeber ZT5 e ZT9. </li><li> Posizionare il T-labirinto sul pad martellante, e stringere la clip vite così tegli ascensore è stabile. </li><li> Trasferire linea risponditore in un test flacone fresco. Snap questo flaconcino contenente linea risponditore sulla parte superiore della T-labirinto. </li><li> Inclinare l&#39;apparecchio T-labirinto e toccare sul tappeto martellante in modo che i moscerini della frutta caduta in ascensore. </li><li> Spostare la parte superiore del l&#39;ascensore in modo in linea sono tra la parte superiore e inferiore del T-labirinto, ma attenzione per mantenere l&#39;ascensore sopra del punto scelta - per evitare le mosche fuoriuscita. </li><li> Ottenere fiale con DSO (vedi sezione 5 per generare fiale DSO-libero che in precedenza occupati da mosche). <ol><li> Mentre le mosche risponditore stanno adeguando a questo nuovo ambiente, mettere in linea emettitore in un flaconcino di prova. </li><li> Posizionare un pezzo di cotone per chiudere la fiala in modo che i moscerini non sfuggono. </li><li> Meccanicamente Agitare questo flaconcino su una mini-vortice come segue: vortex per 15 secondi, togliere il flacone dal vortice per 5 sec. Ripetere 3 volte per un totale di 55 sec. </li><li> Remil sobrio linea emettitore dal flaconcino fresca - trasferendo in una fiala alimentare - e rapidamente posizionare questo DSO fiala riempita in uno dei due lati dell&#39;apparato T-labirinto. </li><li> Alternare il lato di posizionamento per il flacone DSO compilato in ogni nuova esecuzione. </li></ol></li><li> Inserire un test flaconcino fresca sul lato della T-labirinto. </li><li> Portare l&#39;ascensore fino in fondo, in modo che i moscerini della frutta possono scegliere tra il flacone di prova fresco e la fiala test con DSO, e avviare il timer. </li><li> Lasciate che i mosche scegliere tra la fiala fresco e la fiala DSO, per 1 minuto, se non diversamente specificato. </li><li> Dopo 1 min, spostare l&#39;ascensore fino a separare le mosche nel flacone fresco, DSO flacone, e quelli bloccati in ascensore. </li><li> Contare il numero di mosche in ogni fiala, e in ascensore. </li><li> Ripetere questa operazione per ciascun genotipo / condizione. </li></ol> 4. Analizzare i dati sociale Prevenzione <ol><li> Contare il numero di moscerini della frutta in ogni fiala per ogni genotipo, e trasferire i dati in un foglio di calcolo. </li><li> Utilizzando un foglio di calcolo, calcolare il Performance Index (PI) per ciascun genotipo sottraendo il numero di mosche in flaconcino DSO dal numero di mosche in flacone DSO-libera, e dividendo per il numero totale di mosche. </li><li> Confrontare Pis &quot;: mediante software di analisi statistica per l&#39;analisi dei risultati. <ol><li> Idealmente, preparare 2-3 ripetizioni interne per ogni esperimento, e 3 ripetizioni indipendenti, effettuate in giorni diversi, con diverse bottiglie o croci 30. </li><li> Utilizzare test statistici distribuzione gaussiana, come i dati seguono una distribuzione normale. </li><li> Durante il test di un effetto (cioè, densità - figura 2), utilizzare un ANOVA, con post-test di confronto multiplo (colonna per colonna), o un semplice t-test per confrontare ogni risultato a un controllo. </li><li> Quando gli esperimenti di gruppo vengono eseguiti, testare entrambe le condizioni e generi (flaconi vuoti, ha sottolineato und vola non-ha sottolineato; ed effetto genere - figura 1), utilizzando una ANOVA a due vie. </li></ol></li></ol> 5. Generare fiale precedentemente occupato da mosche non accentate - Eseguire Invece di 3,5-3,6 <ol><li> Utilizzare fototassi preparare fiale occupati da mosche che non sono stati sottolineati, come controllo. </li><li> Trasferire le mosche in un flaconcino di prova, e farlo scattare in dell&#39;apparato controcorrente, con fiale freschi vuoti nella posizione di fronte. </li><li> Posare l&#39;apparecchio orizzontalmente con la fiala distale direttamente di fronte a 15 W fluorescente bianca fredda, e coprire con un panno nero. </li><li> Lasciate linea si muovono delicatamente nel flaconcino fresco. Questo potrebbe richiedere alcuni minuti. Una volta che la maggior parte delle mosche camminano verso la luce nel flacone distale, avviare il timer per 1 min. </li><li> Rimuovere delicatamente il panno nero, cambiare la posizione della luce, il recupero con il panno nero, e permettono in linea di lasciare il flacone di prova. </li><li> A quel tempo, initiate l&#39;esperimento scelta in 3.2 a 3.5. Salta 3.6 e 3.7. </li><li> Spostare il cursore dell&#39;apparecchiatura, per separare le mosche che liberate il flaconcino di prova dall&#39;altro lato. </li><li> Utilizzare il flacone di prova sgomberato nel T-labirinto subito per l&#39;esperimento scelta, come in 3.7. </li></ol>

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E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+decline+in+behavioral+performance+of+Drosophila+melanogaster+with+age.">Differential decline in behavioral performance of Drosophila melanogaster with age.</a> Mechanisms of Ageing and Development. , 127-647 (2006).</li></ol>

The authors declare that they have no competing financial interests.

Questo protocollo descrive una procedura dettagliata per il saggio evitamento sociale. Canton-S eviterà solo una fiala in che vola in precedenza stato meccanicamente sottolineato, e che il sesso e il numero di responder non influenza che le prestazioni evitamento sociale. Tuttavia, background genetico dei soccorritori ha una grande influenza. Di seguito sono riportati alcuni passaggi critici per l&#39;esecuzione di questo esperimento con successo: 1) il trasferimento sempre in linea 2 ore p...

L&#39;obiettivo di questo metodo è di quantificare facilmente un nuovo aspetto del semplice comportamento sociale in Drosophila melanogaster, indipendente dalla corteggiamento e aggressività. Interazioni sociali sono fondamentali per il corretto sviluppo e la salute delle persone all&#39;interno di una società, così come la funzionalità di un gruppo sociale nel suo complesso. La complessità di queste interazioni richiede campioni di grandi dimensioni e di un sistema che consente una semplificazione del comportamento come basi genetiche e neurali del comportamento sociale sono ancora poco conosciuti. Drosophila melanogaster è un modello genetico potente che può essere utilizzato per identificare il genetica e basi neurali delle interazioni sociali. Infatti, D. melanogaster ha un repertorio di comportamenti sociali complessi e alcune misure dirette di socializzazione sono già stato fatto 1-7. Tuttavia, la maggior parte di questi sforzi sono stati concentrati sui comportamenti sociali relativamente complessi, come aggresinterazioni sive 3,6, vari aspetti del corteggiamento 3,8-12, e come i social esperienza influisce altri comportamenti quali l&#39;apprendimento, o il ritmo circadiano 13-17. Inoltre, molti di questi test si basano su analisi di modelli di interazione complesse di gruppi di mosche, con il monitoraggio di video e software per analizzare l&#39;abbondanza risultante dei dati. Tali analisi hanno un valore inestimabile, e portare a nuove importanti informazioni quali la dinamica delle interazioni fly-fly in gruppi 7. Un limite, però, è l&#39;inaccessibilità di questi test per la comunità in generale, e la limitata conoscenza dei meccanismi alla base il riconoscimento degli altri. In altre parole, la base dell&#39;emissione di un segnale da un individuo e il suo riconoscimento da un altro è ancora scarsamente compreso 18. Al contrario, vola anche mostrare un comportamento semplice, evitamento sociale, in cui gli individui si muovono da un segnale emesso da mosche sollecitate: la D. Melanogaster stress odorizzante o DSO 19. In un saggio ad alta prestazione, questo comportamento può essere quantificato come la prevenzione di un segnale di stress emesso da altre mosche, o evitamento sociale 19. Mosche sono posti in un apparecchio T-labirinto e data la scelta per evitare una fiala contenente DSO. Utilizzando questo test, CO 2 ha dimostrato di essere un componente del DSO, e parte del circuito neuronale necessario per rispondere a CO 2 è stato dissezionato 19. Il saggio evitamento sociale qui presentato è simile concettualmente ai semplici test comportamentali sviluppati nel laboratorio di Seymour Benzer che hanno permesso generazioni di ricercatori per analizzare comportamenti complessi 20. Analisi di evitamento sociale può essere effettuata conveniente utilizzando il test T-maze, consentendo un più ampio studio del comportamento sociale. Ad esempio, utilizzando questo test abbiamo recentemente dimostrato che i diversi rischi genetici per l&#39;autismo hanno effetti contrastanti in b socialesaggi ehavior. Mutanti di un gene candidato per l&#39;autismo - neurobeachin 21,22 - carenze presenti sia nello spazio sociale (descritte altrove 23) e l&#39;evitare sociale 24. Segnalazione dopaminergico anomala si propone anche di svolgere un ruolo nell&#39;eziologia dell&#39;autismo negli esseri umani 25,26. In contrasto con i risultati ottenuti con neurobeachin, abbiamo scoperto che le prestazioni evitamento sociale è stata influenzata da un aumento o diminuzione dei livelli di Drosophila vescicolare Monoamino Transporter (VMAT) in cellule dopaminergiche, anche se lo spazio sociale è direttamente correlata a questi livelli di VMAT 27. I risultati ottenuti con contrasto neurobeachin e VMAT sottolineano la possibilità di individuare varie forme di comportamento asociale, e quindi le differenti circuiterie neurali sottostanti modulano la risposta ad altri.

<table><tbody><tr><td>Stereo Zoom Microscope&amp;nbsp;</td><td>Nikon&amp;nbsp;&amp;nbsp;</td><td>SMZ-645</td><td>Any other standard scope for fly handling would work</td></tr><tr><td>Small paint brushes&amp;nbsp;</td><td></td><td></td><td>for pushing flies</td></tr><tr><td>Porous Polyethylene, 12&quot; x 12&quot; Sheet</td><td>Flystuff - Genesee</td><td>46-100</td><td>http://www.flystuff.com/ProductInfo.php?productID=46-100&lt;br/&gt; Porous Plastic sheet for the cold anesthesia box</td></tr><tr><td>Mini-Alarm Timer/Stopwatch</td><td></td><td></td><td></td></tr><tr><td>Sharpie pens</td><td></td><td></td><td></td></tr><tr><td>Adhesive Tape</td><td></td><td></td><td></td></tr><tr><td>Mini vortex</td><td>Fisher</td><td>14-955-151&amp;nbsp;</td><td>http://www.fishersci.com/ecomm/servlet/itemdetail&lt;br/&gt; For mechanical agitation of the flies - any vortex would work.</td></tr><tr><td>Corning Life Sciences DL No.:352017, Falcon test tube; round bottom; disposable; no closure, 14 ml; 17 x 100 mm</td><td>Fisher</td><td>14-959-8 &amp;nbsp;&amp;nbsp;</td><td>http://www.fishersci.com/ecomm/servlet/itemdetail?storeId=10652&amp;amp;langId=-1&amp;amp;catalogId=29104&amp;amp;productId=2771811&amp;amp;distype=0&amp;amp;highlightProductsItemsFlag=Y&amp;amp;fromSearch=1&amp;amp;searchType=PROD&amp;amp;hasPromo=0&lt;br/&gt; These snap in place in the in-house made T-maze and counter-current apparatus (see text)</td></tr><tr><td>cotton balls</td><td></td><td></td><td>to close the vials after the experiment.</td></tr><tr><td>trifold board and white bench cover</td><td></td><td></td><td>to provide a white background, and a homogeneous light.</td></tr><tr><td>white bench cover</td><td></td><td></td><td></td></tr><tr><td>pounding pad</td><td></td><td></td><td>any mouse pad works.</td></tr><tr><td>large black cloth</td><td></td><td></td><td>to cover the counter-current apparatus in phototaxis response.</td></tr><tr><td>cool-white light&amp;nbsp;</td><td>Home Depot</td><td>1000516563</td><td>http://www.homedepot.ca/product/illume-26-fluorescent-plug-in-linear/911423&lt;br/&gt; any similar linear light with fluorescent light bulb cool-white at 13-15 W would work</td></tr></tbody></table>

straightforward assay for quantification of social avoidance in drosophila melanogaster

Drosophila melanogaster is an emerging model to study different aspects of social interactions. For example, flies avoid areas previously occupied by stressed conspecifics due to an odorant released during stress known as the Drosophila stress odorant (dSO). Through the use of the T-maze apparatus, one can quantify the avoidance of the dSO by responder flies in a very affordable and robust assay. Conditions necessary to obtain a strong performance are presented here. A stressful experience is necessary for the flies to emit dSO, as well as enough emitter flies to cause a robust avoidance response to the presence of dSO. Genetic background, but not their group size, strongly altered the avoidance of the dSO by the responder flies. Canton-S and Elwood display a higher performance in avoiding the dSO than Oregon and Samarkand strains. This behavioral assay will allow identification of mechanisms underlying this social behavior, and the assessment of the influence of genes and environmental conditions on both emission and avoidance of the dSO. Such an assay can be included in batteries of simple diagnostic tests used to identify social deficiencies of mutants or environmental conditions of interest.

Il saggio evitamento sociale è una prova robusta quantificare la capacità di Drosophila melanogaster riconoscere un segnale di stress (DSO) emessa da altri mosche, e valutare quindi un aspetto delle interazioni sociali. Il test viene eseguito utilizzando un apparecchio comunemente usato in vari saggi comportamento noto come T-labirinto, che presentano in linea con una scelta tra due diverse opzioni - a sinistra oa destra 19,28-31. In questo caso, l&#39;efficienza con cui linea evitano il DSO è qua...

Watch this Scientific Journal Video about Straightforward Assay for Quantification of Social Avoidance in Drosophila melanogaster at JoVE.com

Straightforward Assay for Quantification of Social Avoidance in Drosophila melanogaster

Here, we present a protocol to quantify the avoidance of stressed individuals. This paradigm is powerful yet user-friendly and can be used to assess the influence of genes and environment on one kind of social interaction in Drosophila melanogaster.

Assay Semplice per la quantificazione della Social Prevenzione in<em&gt; Drosophila melanogaster</em

Drosophila melanogaster is an emerging model to study different aspects of social interactions. For example, flies avoid areas previously occupied by ...

straightforward-assay-for-quantification-social-avoidance-drosophila

Universidad de Cartagena UDEC

Research

JoVE Journal

Neuroscience

Straightforward Assay for Quantification of Social Avoidance in Drosophila melanogaster

11.2K Views.  Yale University. Here, we present a protocol to quantify the avoidance of stressed individuals. This paradigm is powerful yet user-friendly and can be used to assess the influence of genes and environment on one kind of social interaction in Drosophila melanogaster.

Video: Straightforward Assay for Quantification of Social Avoidance in Drosophila melanogaster

Assaying Locomotor, Learning, and Memory Deficits in Drosophila Models of Neurodegeneration

Advances in genetic methods have enabled the study of genes involved in human neurodegenerative diseases using Drosophila as a model system1. Most of these diseases, including Alzheimer's, Parkinson's and Huntington's disease are characterized by age-dependent deterioration in learning and memory functions and movement coordination2. Here we use behavioral assays, including the negative geotaxis assay3 and the aversive phototaxic suppression assay (APS assay)4,5, to show that some of the behavior characteristics associated with human neurodegeneration can be recapitulated in flies. In the negative geotaxis assay, the natural tendency of flies to move against gravity when agitated is utilized to study genes or conditions that may hinder locomotor capacities. In the APS assay, the learning and memory functions are tested in positively-phototactic flies trained to associate light with aversive bitter taste and hence avoid this otherwise natural tendency to move toward light. Testing these trained flies 6 hours post-training is used to assess memory functions. Using these assays, the contribution of any genetic or environmental factors toward developing neurodegeneration can be easily studied in flies.

Assaying Locomotor, Learning, and Memory Deficits in Drosophila Models of Neurodegeneration

Behavior assays for measuring locomotor functions, learning, and memory abilities in Drosophila.

Deficit Locomotore dosaggio, apprendimento e memoria in Drosophila Modelli di neurodegenerazione

Advances in genetic methods have enabled the study of genes involved in human neurodegenerative diseases using Drosophila as a model system1. Most of ...

Ricerca

Neuroscienze

Methods to Assay Drosophila Behavior

Drosophila melanogaster, the fruit fly, has been used to study molecular mechanisms of a wide range of human diseases such as cancer, cardiovascular disease and various neurological diseases1. We have optimized simple and robust behavioral assays for determining larval locomotion, adult climbing ability (RING assay), and courtship behaviors of Drosophila. These behavioral assays are widely applicable for studying the role of genetic and environmental factors on fly behavior. Larval crawling ability can be reliably used for determining early stage changes in the crawling abilities of Drosophila larvae and also for examining effect of drugs or human disease genes (in transgenic flies) on their locomotion. The larval crawling assay becomes more applicable if expression or abolition of a gene causes lethality in pupal or adult stages, as these flies do not survive to adulthood where they otherwise could be assessed. This basic assay can also be used in conjunction with bright light or stress to examine additional behavioral responses in Drosophila larvae. Courtship behavior has been widely used to investigate genetic basis of sexual behavior, and can also be used to examine activity and coordination, as well as learning and memory. Drosophila courtship behavior involves the exchange of various sensory stimuli including visual, auditory, and chemosensory signals between males and females that lead to a complex series of well characterized motor behaviors culminating in successful copulation. Traditional adult climbing assays (negative geotaxis) are tedious, labor intensive, and time consuming, with significant variation between different trials2-4. The rapid iterative negative geotaxis (RING) assay5 has many advantages over more widely employed protocols, providing a reproducible, sensitive, and high throughput approach to quantify adult locomotor and negative geotaxis behaviors. In the RING assay, several genotypes or drug treatments can be tested simultaneously using large number of animals, with the high-throughput approach making it more amenable for screening experiments.

Methods to Assay Drosophila Behavior

Drosophila melanogaster is a genetically and behaviorally tractable model system that has been used to understand the molecular and cellular basis of many important biological processes for over a century 1. Drosophila has been well exploited to gain insights into the genetic basis of fly behavior.

Metodi per test Drosophila Comportamento

Drosophila melanogaster, the fruit fly, has been used to study molecular mechanisms of a wide range of human diseases such as cancer, cardiovascular ...

A Single-fly Assay for Foraging Behavior in Drosophila

For many animals, hunger promotes changes in the olfactory system in a manner that facilitates the search for appropriate food sources. In this video article, we describe an automated assay to measure the effect of hunger or satiety on olfactory dependent food search behavior in the adult fruit fly Drosophila melanogaster. In a light-tight box illuminated by red light that is invisible to fruit flies, a camera linked to custom data acquisition software monitors the position of six flies simultaneously. Each fly is confined to walk in individual arenas containing a food odor at the center. The testing arenas rest on a porous floor that functions to prevent odor accumulation. Latency to locate the odor source, a metric that reflects olfactory sensitivity under different physiological states, is determined by software analysis. Here, we discuss the critical mechanics of running this behavioral paradigm and cover specific issues regarding fly loading, odor contamination, assay temperature, data quality, and statistical analysis.

A Single-fly Assay for Foraging Behavior in Drosophila

In this video article, we describe an automated assay to measure the effect of hunger or satiety on olfactory dependent food search behavior in the adult fruit fly Drosophila melanogaster.

A Single-fly saggio per comportamento alimentare in<em&gt; Drosophila</em

For many animals, hunger promotes changes in the olfactory system in a manner that facilitates the search for appropriate food sources. In this video ...

Drosophila Passive Avoidance Behavior as a New Paradigm to Study Associative Aversive Learning

This protocol describes a new paradigm for analyzing aversive associative learning in adult flies (Drosophila melanogaster). The paradigm is analogous to passive avoidance behavior in laboratory rodents in which animals learn to avoid a compartment where they have previously received an electric shock. The assay takes advantage of negative geotaxis in flies, which manifests as an urge to climb up when they are placed on a vertical surface. The setup consists of vertically oriented upper and lower compartments. On the first trial, a fly is placed into a lower compartment from where it usually exits&#160;within 3-15 s, and steps into the upper compartment where it receives an electric shock. During the second trial, 24 h later, the latency is significantly increased. At the same time, the number of shocks is decreased compared to the first trial, indicating that flies formed long-term memory about the upper compartment. The recordings of latencies and number of shocks could be performed with a tally counter and a stopwatch or with an Arduino-based simple device. To illustrate how the assay can be used, the passive avoidance behavior of D. melanogaster and D. simulans male and female were characterized here. Comparison of latencies and number of shocks revealed that both D. melanogaster and D. simulans flies efficiently learned the passive avoidance behavior. No statistical differences were observed between male and female flies. However, males were a little faster while entering the upper compartment on the first trial, while females received a slightly higher number of shocks in every retention trial. The Western diet (WD) significantly impaired learning and memory in male flies while flight exercise counterbalanced this effect. Taken together, the passive avoidance behavior in flies offers a simple and reproducible assay that could be used for studying basic mechanisms of learning and memory.

Drosophila Passive Avoidance Behavior as a New Paradigm to Study Associative Aversive Learning

This work describes a simple behavioral paradigm that allows the analysis of aversive associative learning in adult fruit flies. The method is based on suppressing the innate negative geotaxis behavior due to the association formed between a specific environmental context and an electric shock.

Drosofila Comportamento di evitamento passivo come nuovo paradigma per studiare l'apprendimento avversivo associativo

This protocol describes a new paradigm for analyzing aversive associative learning in adult flies (Drosophila melanogaster). The paradigm is analogous to ...

Comportamento

A Simple Technique to Assay Locomotor Activity in Drosophila

Drosophila melanogaster is an ideal model organism for studying various diseases due to its abundance of advanced genetic manipulation techniques and diverse behavioral features. Identifying behavioral deficiency in animal models is a crucial measure of disease severity, for example, in neurodegenerative diseases where patients often experience impairments in motor function. However, with the availability of various systems to track and assess motor deficits in fly models, such as drug-treated or transgenic individuals, an economical and user-friendly system for precise evaluation from multiple angles is still lacking. A method based on the AnimalTracker application programming interface (API) is developed here, which is compatible with the Fiji image processing program, to systematically evaluate the movement activities of both adult and larval individuals from recorded video, thus allowing for the analysis of their tracking behavior. This method requires only a high-definition camera and a computer peripheral hardware integration to record and analyze behavior, making it an affordable and effective approach for screening fly models with transgenic or environmental behavioral deficiencies. Examples of behavioral tests using pharmacologically treated flies are given to show how the techniques can detect behavioral changes in both adult flies and larvae in a highly repeatable manner.

A Simple Technique to Assay Locomotor Activity in Drosophila

The present protocol assesses the locomotor activity of Drosophila by tracking and analyzing the movement of flies in a hand-made arena using open-source software Fiji, compatible with plugins to segment pixels of each frame based on high-definition video recording to calculate parameters of speed, distance, etc.

Una semplice tecnica per analizzare l'attività locomotoria nella Drosophila

Drosophila melanogaster is an ideal model organism for studying various diseases due to its abundance of advanced genetic manipulation techniques and ...

Conditions Affecting Social Space in Drosophila melanogaster

The social space assay described here can be used to quantify social interactions of Drosophila melanogaster &#8212; or other small insects &#8212; in a straightforward manner. As we previously demonstrated 1, in a two-dimensional chamber, we first force the flies to form a tight group, subsequently allowing them to take their preferred distance from each other. After the flies have settled, we measure the distance to the closest neighbor (or social space), processing a static picture with free online software (ImageJ). The analysis of the distance to the closest neighbor allows researchers to determine the effects of genetic and environmental factors on social interaction, while controlling for potential confounding factors. Diverse factors such as climbing ability, time of day, sex, and number of flies, can modify social spacing of flies. We thus propose a series of experimental controls to mitigate these confounding effects. This assay can be used for at least two purposes. First, researchers can determine how their favorite environmental shift (such as isolation, temperature, stress or toxins) will impact social spacing 1,2. Second, researchers can dissect the genetic and neural underpinnings of this basic form of social behavior 1,3. Specifically, we used it as a diagnostic tool to study the role of orthologous genes thought to be involved in social behavior in other organisms, such as candidate genes for autism in humans 4.

Conditions Affecting Social Space in Drosophila melanogaster

The effect of genes and environment on social space of Drosophila melanogaster can be quantified through a powerful but straightforward analytical paradigm. We show here different factors that can affect this social space, and thus need to be taken into consideration when designing experiments in this paradigm.

Condizioni che influiscono spazio sociale in<em&gt; Drosophila melanogaster</em

The social space assay described here can be used to quantify social interactions of Drosophila melanogaster &#8212; or other small insects &#8212; in a ...

High Throughput Assay to Examine Egg-Laying Preferences of Individual Drosophila melanogaster

Recently, egg-laying preference of Drosophila has emerged as a genetically tractable model to study the neural basis of simple decision-making processes. When selecting sites to deposit their eggs, female flies are capable of ranking the relative attractiveness of their options and choosing the &#34;greater of two goods.&#34; However, most egg-laying preference assays are not practical if one wants to take a systematic genetic screening approach to search for the circuit basis underlying this simple decision-making process, as they are population-based and laborious to set up. To increase the throughput of studying of egg-laying preferences of single females, we developed custom chambers that each can simultaneously assay egg-laying preferences of up to thirty individual flies as well as a protocol that ensures each female has a high egg-laying rate (so that their preference is readily discernable and more convincing). Our approach is simple to execute and produces very consistent results. Additionally, these chambers can be equipped with different attachments to allow video recording the egg-laying animals and to deliver light for optogenetics studies. This article provides the blueprints for fabricating these chambers and the procedure for preparing the flies to be assayed in these chambers.

High Throughput Assay to Examine Egg-Laying Preferences of Individual Drosophila melanogaster

This protocol describes a high throughput assay for testing egg-laying preferences of Drosophila melanogaster at single-animal resolution. This assay provides a simple, efficient, and scalable platform to identify genes and circuit components that control a simple decision-making process.

High Throughput test per esaminare Preferenze deposizione delle uova dei singoli<em&gt; Drosophila melanogaster</em

Recently, egg-laying preference of Drosophila has emerged as a genetically tractable model to study the neural basis of simple decision-making processes. ...

Drosofila Modelli per i tratti genetici e comportamentali del disturbo dello spettro autistico

Paradigms for Behavioral Assessment in Drosophila Model of Autism Spectrum Disorder

Autism Spectrum Disorder (ASD) encompasses a heterogeneous group of neurodevelopmental disorders with common behavioral symptoms including deficits in social interaction and ability for communication, enhanced restricted or repetitive behaviors, and also, in some cases, learning disability and motor deficit. Drosophila has served as an unparalleled model organism for modeling a great number of human diseases. As many genes have been implicated in ASD, fruit flies have emerged as a powerful and efficient way to test the genes putatively involved with the disorder. As hundreds of genes, with varied functional roles, are implicated in ASD, a single genetic fly model of ASD is unfeasible; instead, individual genetic mutants, gene knockdowns, or overexpression-based studies of the fly homologs of ASD-associated genes are the common means for gaining insight regarding molecular pathways underlying these gene products. A host of behavioral techniques are available in Drosophila which provide easy readout of deficits in specific behavioral components. Social space assay and aggression and courtship assays in flies have been shown to be useful in assessing defects in social interaction or communication. Grooming behavior in flies is an excellent readout of repetitive behavior. Habituation assay is used in flies to estimate the ability for habituation learning, which is found to be affected in some ASD patients. A combination of these behavioral paradigms can be utilized to make a thorough assessment of the human ASD-like disease state in flies. Using Fmr1 mutant flies, recapitulating Fragile-X syndrome in humans, and POGZ-homolog row knockdown in fly neurons, we have shown quantifiable deficits in social spacing, aggression, courtship behavior, grooming behavior, and habituation. These behavioral paradigms are demonstrated here in their simplest and straightforward forms with an assumption that it would facilitate their widespread use for research on ASD and other neurodevelopmental disorders in fly models.

Autore in primo piano: Esplorazione dei sintomi del disturbo dello spettro autistico nei moscerini della frutta: modelli genetici e test comportamentali

Autism Spectrum Disorder (ASD) is associated with impaired social and communicative behavior and the emergence of repetitive behavior. For studying the interrelation between ASD genes and behavioral deficits in the Drosophila model, five behavioral paradigms are described in this paper for assaying social spacing, aggression, courtship, grooming, and habituation behavior.

Paradigmi per la valutazione comportamentale nel modello di Drosophila del disturbo dello spettro autistico

Autism Spectrum Disorder (ASD) encompasses a heterogeneous group of neurodevelopmental disorders with common behavioral symptoms including deficits in ...

Assay Semplice per la quantificazione della Social Prevenzione in

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High Throughput test per esaminare Preferenze deposizione delle uova dei singoli

Paradigmi per la valutazione comportamentale nel modello di Drosophila del disturbo dello spettro autistico

Paradigmi per la valutazione comportamentale nel modello di Drosophila del disturbo dello spettro autistico

Paradigmi per la valutazione comportamentale nel modello di Drosophila del disturbo dello spettro autistico