Ringraziamo il Bloomington Drosophila Stock Center per gli stock fly; C. Gahr, JT Alkema e S. van Hasselt per il loro primo tentativo di sviluppare il dosaggio dell&#39;aria sotto pressione; Jasper Bosman per il consiglio sulla coltivazione del lievito; E Rezza Azanchi e Joel Levine per sviluppare in origine il monitoraggio del comportamento di accoppiamento Drosophila . JA Gorter è stato sostenuto da una borsa di studio della Neuroscience Research School di BCN / NWO. Questo lavoro è stato sostenuto in parte dall&#39;organizzazione olandese per la ricerca scientifica (NWO) (riferimento: 821.02.020) a JC Billeter.

<ol>
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C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurogenetics+of+female+reproductive+behaviors+in+Drosophila+melanogaster.">Neurogenetics of female reproductive behaviors in Drosophila melanogaster.</a> BS:ADGEN. 85, 1-108 (2014).</li><li>Liu, H., Kubli, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sex-peptide+is+the+molecular+basis+of+the+sperm+effect+in+Drosophila+melanogaster.">Sex-peptide is the molecular basis of the sperm effect in Drosophila melanogaster.</a> Proc. Natl. Acad. Sci. U S A. 100 (17), 9929-9933 (2003).</li><li>Ram, K. R., Wolfner, M. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sustained+post-mating+response+in+Drosophila+melanogaster+requires+multiple+seminal+fluid+Proteins.">Sustained post-mating response in Drosophila melanogaster requires multiple seminal fluid Proteins.</a> PLoS gen. 3 (12), 2428-2438 (2007).</li><li>Yapici, N., Kim, Y. -. J., Ribeiro, C., Dickson, 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=A+receptor+that+mediates+the+post-mating+switch+in+Drosophila+reproductive+behaviour.">A receptor that mediates the post-mating switch in Drosophila reproductive behaviour.</a> Nature. 451 (7174), 33-37 (2008).</li><li>Yang, C. -. H., 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=Control+of+the+postmating+behavioral+switch+in+Drosophila+females+by+internal+sensory+neurons.">Control of the postmating behavioral switch in Drosophila females by internal sensory neurons.</a> Neuron. 61 (4), 519-526 (2009).</li><li>H&#228;semeyer, M., Yapici, N., Heberlein, U., Dickson, 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=Sensory+neurons+in+the+Drosophila+genital+tract+regulate+female+reproductive+behavior.">Sensory neurons in the Drosophila genital tract regulate female reproductive behavior.</a> Neuron. 61 (4), 511-518 (2009).</li><li>Rez&#225;val, C., Pavlou, H. J., Dornan, A. J., Chan, Y. -. B., Kravitz, E. A., Goodwin, S. 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Questo protocollo descrive un test per verificare il comportamento di accoppiamento per 24 ore mentre controlla continuamente i segnali ambientali che una coppia di accoppiamenti è ipotizzata per utilizzare per determinare la frequenza di accoppiamento. È possibile aumentare la frequenza di accoppiamento in risposta all&#39;aria del lievito erogata attraverso un sistema di aria compressa quando il mezzo contiene anche il lievito ( Figura 2B ). Inoltre, una risposta simile in frequenza di accoppiamento...

I comportamenti riproduttivi hanno generalmente elevati costi energetici, soprattutto per le femmine, che producono gameti più grandi rispetto ai maschi e devono scegliere con cura le condizioni per aumentare la loro prole in via di sviluppo. A causa del costo energetico, non sorprende che la riproduzione sia connessa alle condizioni nutrizionali. Questo è vero nella maggior parte, se non in tutti, animali compresi i mammiferi, la cui pubertà può essere ritardata dalla malnutrizione e il cui comportamento sessuale può essere influenzato negativamente dalla restrizione alimentare 1 . La riproduzione dell&#39;organismo del modello genetico Drosophila melanogaster è anche influenzata da condizioni nutrizionali. La corte dei maschi a livello più elevato in presenza di volatili alimentari 2 e le femmine sono più ricettive sessualmente in presenza di lieviti, un importante nutrimento per la produzione di uova e la sopravvivenza dei figli 3 , 4 , 5 . QuestoLa risposta riproduttiva evoluzionistica riposta all&#39;alimentazione offre l&#39;opportunità di studiare meccanismi che connettono la disponibilità di cibo ambientale alla riproduzione sessuale in un organismo geneticamente tractable e time-efficient. Infatti, il lavoro in D. melanogaster ha implicato il percorso dell&#39;insulina come un importante regolatore della connessione tra il comportamento alimentare e l&#39;accoppiamento 6 . Ha anche dimostrato che l&#39;atto di accoppiamento modifica la preferenza alimentare delle femmine nonché i neuroni chemiosensori associati 7 , 8 , 9 . È chiaro che le indicazioni alimentari influenzano i comportamenti riproduttivi in D. melanogaster . Questi effetti sembrano interessare principalmente le femmine, in particolare quelle che hanno già maturato 5 . Tuttavia, per testare questi effetti acuti delle condizioni ambientali, il dosaggio classico utilizzato per il comportamento femminile di accoppiamento potrebbeNon essere molto adatto a causa delle lunghe interruzioni tra gli episodi di accoppiamento. Nel classico test di rimozione, una femmina vergine prima si collega con un maschio e viene immediatamente isolata e presentata con un nuovo maschio 24-48 h più tardi. Questo saggio classico è stato usato con grande successo per identificare componenti dell&#39;eiaculato maschile che modificano il comportamento femminile e la risposta femminile 12 , 13 , 14 , 15 , 16 , 17 , 18 . Il test di accoppiamento continuo dimostrato qui è quindi un&#39;aggiunta a saggi classici di accoppiamento che possono essere utilizzati per studiare l&#39;effetto acuto delle condizioni ambientali sui comportamenti riproduttivi. Utilizzando il dosaggio continuo per il comportamento di accoppiamento che viene spiegato qui, abbiamo precedentemente mostrato che una coppia di mosche esposte al lievito rimateràSempre in un periodo di osservazione di 24 ore 5 , 19 , 20 , 21 , mentre mosche non esposte al cibo rimarrà solo una volta 5 . Questa scoperta può essere sconcertante alla luce di una grande parte della letteratura D. melanogaster che indica che le femmine non si ritraggono per diversi giorni dopo un accoppiamento iniziale (riveduto nei riferimenti 10 , 11 ). Tuttavia, questa discrepanza può essere facilmente spiegata da condizioni di analisi, in cui una femmina è isolata per uno a diversi giorni prima di fornire una nuova opportunità di accoppiamento. Se la coppia non si accoppia in questo periodo di osservazione, la donna è caratterizzata come non ricettivo. Inoltre, la frequenza di accoppiamento elevato non dovrebbe sorprendere, dato che i dati provenienti da mosche selvatiche mostrano che le femmine contengono spermatozoi da 4 a 6 maschi nei loro organi di stoccaggio; Così inDicating che le femmine naturalmente rimane diverse volte 22 , 23 . Qui dimostriamo l&#39;uso di questo saggio di accoppiamento continuo per scoprire come le mosche raccolgono e combinano informazioni sulle condizioni ambientali per modulare la frequenza di accoppiamento. Questo test consente di testare un numero relativamente elevato di coppie di maternità per studi genetici e di verificare l&#39;influenza dei segnali ambientali volatili e non volatili. Il dosaggio viene tipicamente eseguito per 24 h, ma può essere esteso a 48 h, consentendo la sperimentazione di segnali ambientali di ciclo come il ciclo scuro (LD). Ci dimostriamo questo test verificando l&#39;influenza dei segnali volatili da una coltura di lieviti all&#39;interno di un sistema di aria pressurata in combinazione con la disponibilità di nutrienti di lievito non volatili nel substrato alimentare. L&#39;impianto a pressione pressurizzata pompava continuamente segnali volatili in un&#39;arena di accoppiamento che contengonoSubstrato alimentare e una coppia di test (il cui comportamento di accoppiamento è monitorato). Per determinare ulteriormente le specificità attraverso le quali il lievito influenza l&#39;accoppiamento, si prova un importante composto volatile di lievito, cioè acido acetico 24 , in combinazione con un contenuto di amminoacidi che corrisponde a quello del lievito nel substrato alimentare, sotto forma di peptone (amino Acidi derivati ​​dalla digestione enzimatica delle proteine ​​animali). Insieme questi esperimenti dimostrano come l&#39;effetto dei segnali ambientali sul comportamento di accoppiamento di D. melanogaster possa essere testato con questo saggio.

<table><tbody><tr><td>&lt;strong&gt;Cabinet&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Stainless steel kitchen cabinet</td><td>Horecaworld</td><td>7412.0105</td><td></td></tr><tr><td>White LEDs</td><td>Lucky Light</td><td>ll-583wc2c-001</td><td>Cold white, 20 mAmp and 2 V</td></tr><tr><td>Red LEDs</td><td>Lucky Ligt</td><td>ll-583vc2c-v1-4da</td><td>Wavelength between 625 nm, 20 mAmp and 6 V</td></tr><tr><td>Resistor</td><td>Royal Ohm</td><td>CFR0W4J0561A50</td><td>560 ohm, 0.25 W, 250 V and 5 % tolerance</td></tr><tr><td>Smartphone light meter app</td><td>Patrick Giudicelli</td><td></td><td>Light/Lux Meter FREE, version 1.1.1</td></tr><tr><td>Power timer</td><td>Alecto</td><td>TS-121</td><td></td></tr><tr><td>Metal brackets</td><td></td><td></td><td>Sharp angle 5 by 5 mm,&amp;nbsp; 2 x 5450 and 1 x 1100 mm long</td></tr><tr><td>Frosted glass plate</td><td></td><td></td><td>1190 x 545 x 5 mm</td></tr><tr><td>Filter paper sheets</td><td>LEE filters</td><td>220</td><td>White frost</td></tr><tr><td>Small fan</td><td>Nanoxia Deep silence</td><td>4260285292828</td><td>80 mm Ultra-Quiet PC Fan, 1200 RPM</td></tr><tr><td>Big fan</td><td>Nanoxia Deep silence</td><td>4260285292910</td><td>120 mm Ultra-Quiet PC Fan, 650-1500 RPM</td></tr><tr><td>Webcam camera</td><td>Logitech</td><td>950270</td><td>B910 HD WEBCAM OEM, Angle: 78-degree, resolution: 5-million-pixel&amp;nbsp;&amp;nbsp;</td></tr><tr><td>Camera software</td><td>DeskShare</td><td></td><td>Security monitor pro</td></tr><tr><td>&lt;strong&gt;Name&lt;/strong&gt;</td><td>&lt;strong&gt;Company&lt;/strong&gt;</td><td>&lt;strong&gt;Catalog Number&lt;/strong&gt;</td><td>&lt;strong&gt;Comments&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;Fly rearing&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Fly rearing bottles</td><td>Flystuff</td><td>32-130</td><td>6oz Drosophila stock bottle</td></tr><tr><td>Flypad</td><td>Flystuff</td><td>59-114</td><td></td></tr><tr><td>Wild-type flies</td><td></td><td></td><td>Canton-S</td></tr><tr><td>Fly rearing vials</td><td>Dominique Dutscher</td><td>789008</td><td>Drosophila tubes narrow 25x95 mm</td></tr><tr><td>Incubator</td><td>Sanyo</td><td>MIR-154</td><td></td></tr><tr><td>Magnetic hot plate</td><td>Heidolph</td><td>505-20000-00</td><td>MR Hei-Standard</td></tr><tr><td>Agar</td><td>Caldic Ingredients B.V.</td><td>010001.26.0</td><td></td></tr><tr><td>Glucose</td><td>Gezond&amp;amp;wel</td><td>1019155</td><td>Dextrose/Druivensuiker</td></tr><tr><td>Sucrose</td><td>Van Gilse</td><td></td><td>Granulated sugar</td></tr><tr><td>Cornmeal</td><td>Flystuff</td><td>62-100</td><td></td></tr><tr><td>Wheat germ</td><td>Gezond&amp;amp;wel</td><td>1017683</td><td></td></tr><tr><td>Soy flour</td><td>Flystuff</td><td>62-115</td><td></td></tr><tr><td>Molasses</td><td>Flystuff</td><td>62-117</td><td></td></tr><tr><td>Active dry yeast</td><td>Red Star</td><td></td><td></td></tr><tr><td>Tegosept</td><td>Flystuff</td><td>20-258</td><td>100%</td></tr><tr><td>Peptone (bacto)</td><td>BD</td><td>211677</td><td></td></tr><tr><td>Acetic Acid</td><td>Merck</td><td>1000631000</td><td>Glacial, 100%</td></tr><tr><td>Small petridish</td><td>Greiner bio-one</td><td>627102</td><td>35 x 10 mm with vents</td></tr><tr><td>Paraffin film</td><td>Bemis NA</td><td></td><td>Parafilm</td></tr><tr><td>&lt;strong&gt;Name&lt;/strong&gt;</td><td>&lt;strong&gt;Company&lt;/strong&gt;</td><td>&lt;strong&gt;Catalog Number&lt;/strong&gt;</td><td>&lt;strong&gt;Comments&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;Yeast and pressurised air set-up&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Big petridish</td><td>Gosselin</td><td>BP140-01</td><td>140 x 20.6 mm</td></tr><tr><td>Ultrapure water</td><td>Millipore corporation</td><td></td><td>MiliQ</td></tr><tr><td>Yeast extract</td><td>BD</td><td>212750</td><td></td></tr><tr><td>Agar 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unit</td><td>Whatman</td><td>10462100</td><td></td></tr><tr><td>Serological pipettes</td><td>VWR</td><td>612-1600</td><td></td></tr><tr><td>Syringe</td><td>BD Plastipak</td><td>300013</td><td></td></tr><tr><td>Hot glue</td><td>Pattex</td><td></td><td></td></tr><tr><td>Syringe filter</td><td>Whatman</td><td></td><td>FP 30/pore size 0.45 mm CA-S</td></tr><tr><td>&lt;strong&gt;Name&lt;/strong&gt;</td><td>&lt;strong&gt;Company&lt;/strong&gt;</td><td>&lt;strong&gt;Catalog Number&lt;/strong&gt;</td><td>&lt;strong&gt;Comments&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;Analysis&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Statistics software</td><td>R</td><td></td><td>lme4 package</td></tr></tbody></table>

a method to test the effect of environmental cues on mating behavior in drosophila melanogaster

L&#39;azionamento sessuale di un individuo è influenzato dal genotipo, dall&#39;esperienza e dalle condizioni ambientali. Come questi fattori interagiscono per modulare i comportamenti sessuali rimane poco compreso. In Drosophila melanogaster , i segnali ambientali, come la disponibilità di cibo, influenzano l&#39;attività di accoppiamento che offre un sistema tracciato per indagare i meccanismi che modulano il comportamento sessuale. In D. melanogaster , i segnali ambientali vengono spesso rilevati attraverso i sistemi gassosi e olfattivi chemosensoriali. Qui espostiamo un metodo per testare l&#39;effetto dei segnali chimici ambientali sul comportamento dell&#39;accoppiamento. Il dosaggio è costituito da una piccola arena di accoppiamento contenente un mezzo alimentare e una coppia di accoppiamenti. La frequenza di accoppiamento per ogni coppia viene monitorata continuamente per 24 ore. Qui esprimiamo l&#39;applicabilità di questo test per testare i composti ambientali da una sorgente esterna attraverso un sistema di aria compressa e la manipolazione dei componenti ambientali direttamente nell&#39;arena di accoppiamento. L&#39;uSe di un sistema di aria compressa è particolarmente utile per testare l&#39;effetto di composti molto volatili, mentre manipolazione di componenti direttamente nell&#39;arena di accoppiamento può essere utile per accertare la presenza di un composto. Questo saggio può essere adattato per rispondere a domande sull&#39;influenza delle indicazioni genetiche e ambientali sul comportamento e sulla fecondità di accoppiamento e su altri comportamenti riproduttivi maschi e femmine.

Utilizzando questo test continuo, il comportamento di accoppiamento e la frequenza di accoppiamento in specifiche possono essere determinati in condizioni ambientali sperimentali. Per controllare le condizioni ambientali, abbiamo trasformato un armadio in acciaio inox in una zona di prova, con una propria sorgente luminosa e diffusione che assicura un&#39;elevata abbondanza di luce e una minima quantità di riflessi dalla parte superiore delle arene di accoppiamento ( figura 1A</...

Watch this Scientific Journal Video about A Method to Test the Effect of Environmental Cues on Mating Behavior in Drosophila melanogaster at JoVE.com

A Method to Test the Effect of Environmental Cues on Mating Behavior in Drosophila melanogaster

Noi dimostriamo un dosaggio per analizzare i suggerimenti ambientali e genetici che influenzano il comportamento di accoppiamento nella mosca di frutta Drosophila melanogaster .

Un metodo per testare l &#39;effetto dei segnali ambientali sul comportamento di accoppiamento in<em&gt; Drosophila melanogaster</em

An individual's sexual drive is influenced by genotype, experience and environmental conditions. How these factors interact to modulate sexual behaviors ...

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Universidad de Cartagena UDEC

Research

JoVE Journal

Behavior

A Method to Test the Effect of Environmental Cues on Mating Behavior in Drosophila melanogaster

9.0K Views.  University of Groningen.  Noi dimostriamo un dosaggio per analizzare i suggerimenti ambientali e genetici che influenzano il comportamento di accoppiamento nella mosca di frutta Drosophila melanogaster . 

Video: A Method to Test the Effect of Environmental Cues on Mating Behavior in Drosophila melanogaster

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 ...

Ricerca

Neuroscienze

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 ...

Methods to Test Endocrine Disruption in Drosophila melanogaster

In recent years there has been growing evidence that all organisms and the environment are exposed to hormone-like chemicals, known as endocrine disruptor chemicals (EDCs). These chemicals may alter the normal balance of endocrine systems and lead to adverse effects, as well as an increasing number of hormonal disorders in the human population or disturbed growth and reduced reproduction in the wildlife species. For some EDCs, there are documented health effects and restrictions on their use. However, for most of them, there is still no scientific evidence in this sense. In order to verify potential endocrine effects of a chemical in the full organism, we need to test it in appropriate model systems, as well as in the fruit fly, Drosophila melanogaster. Here we report detailed in vivo protocols to study endocrine disruption in Drosophila, addressing EDC effects on the fecundity/fertility, developmental timing, and lifespan of the fly. In the last few years, we used these Drosophila life traits to investigate the effects of exposure to 17-&#945;-ethinylestradiol (EE2), bisphenol A (BPA), and bisphenol AF (BPA F). Altogether, these assays covered all Drosophila life stages and made it possible to evaluate endocrine disruption in all hormone-mediated processes. Fecundity/fertility and developmental timing assays were useful to measure the EDC impact on the fly reproductive performance and on developmental stages, respectively. Finally, the lifespan assay involved chronic EDC exposures to adults and measured their survivorship. However, these life traits can also be influenced by several experimental factors that had to be carefully controlled. So, in this work, we suggest a series of procedures we have optimized for the right outcome of these assays. These methods allow scientists to establish endocrine disruption for any EDC or for a mixture of different EDCs in Drosophila, although to identify the endocrine mechanism responsible for the effect, further essays could be needed.

Methods to Test Endocrine Disruption in Drosophila melanogaster

Endocrine disruptor chemicals (EDCs) represent a serious problem for organisms and for natural environments. Drosophila melanogaster represents an ideal model to study EDC effects in vivo. Here, we present methods to investigate endocrine disruption in Drosophila, addressing EDC effects on fecundity, fertility, developmental timing, and lifespan of the fly.

Metodi per testare la disgregazione endocrina nella Drosophila melanogaster

In recent years there has been growing evidence that all organisms and the environment are exposed to hormone-like chemicals, known as endocrine disruptor ...

Biologia dello sviluppo

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.

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 ...

Dyeing Insects for Behavioral Assays: the Mating Behavior of Anesthetized Drosophila

Mating experiments using Drosophila have contributed greatly to the understanding of sexual selection and behavior. Experiments often require simple, easy and cheap methods to distinguish between individuals in a trial. A standard technique for this is CO2 anaesthesia and then labelling or wing clipping each fly. However, this is invasive and has been shown to affect behavior. Other techniques have used coloration to identify flies. This article presents a simple and non-invasive method for labelling Drosophila that allows them to be individually identified within experiments, using food coloring. This method is used in trials where two males compete to mate with a female. Dyeing allowed quick and easy identification. There was, however, some difference in the strength of the coloration across the three species tested. Data is presented showing the dye has a lower impact on mating behavior than CO2 in Drosophila melanogaster. The impact of CO2 anaesthesia is shown to depend on the species of Drosophila, with D. pseudoobscura and D. subobscura showing no impact, whereas D. melanogaster males had reduced mating success. The dye method presented is applicable to a wide range of experimental designs.

Dyeing Insects for Behavioral Assays: the Mating Behavior of Anesthetized Drosophila

This protocol describes a simple, cost effective way to individually identify Drosophila or other insects. Demonstration data investigating mating success across three species of Drosophila show that this method is comparable or better than the use of CO2 anaesthesia.

Tintoria Insetti per comportamentali saggi: il comportamento di accoppiamento di anestetizzati<em&gt; Drosophila</em

Mating experiments using Drosophila have contributed greatly to the understanding of sexual selection and behavior. Experiments often require simple, easy ...

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 ...

A Behavioral Assay for Mechanosensation of MARCM-based Clones in Drosophila melanogaster

Because of the structural and functional homology to the hair cells of the mammalian inner ear, the neurons that innervate the Drosophila external sense organs provide an excellent model system for the study of mechanosensation. This protocol describes a simple touch behavior in fruit flies which can be used to identify mutations that interfere with mechanosensation. The tactile stimulation of a macrochaete bristle on the thorax of flies elicits a grooming reflex from either the first or third leg. Mutations that interfere with mechanotransduction (such as NOMPC), or with other aspects of the reflex arc, can inhibit the grooming response. A traditional screen of adult behaviors would have missed mutants that have essential roles during development. Instead, this protocol combines the touch screen with mosaic analysis with a repressible cell marker (MARCM) to allow for only limited regions of homozygous mutant cells to be generated and marked by the expression of green fluorescent protein (GFP). By testing MARCM clones for abnormal behavioral responses, it is possible to screen a collection of lethal p-element mutations to search for new genes involved in mechanosensation that would have been missed by more traditional methods.

A Behavioral Assay for Mechanosensation of MARCM-based Clones in Drosophila melanogaster

In order to identify novel mutations affecting mechanosensation, we designed an assay that measures the behavioral response to tactile stimulation of fly bristles in mutant clones generated by the MARCM method. The combination of techniques allows for the identification of mechanosensitive mutations that would otherwise be lethal.

Un test comportamentale per Mechanosensation di cloni in marcm-based<em&gt; Drosophila melanogaster</em

Because of the structural and functional homology to the hair cells of the mammalian inner ear, the neurons that innervate the Drosophila external sense ...

A Simple Way to Measure Alterations in Reward-seeking Behavior Using Drosophila melanogaster

We describe a protocol for measuring ethanol self-administration in fruit flies (Drosophila melanogaster) as a proxy for changes in reward states. We demonstrate a simple way to tap into the fly reward system, modify experiences related to natural reward, and use voluntary ethanol consumption as a measure for changes in reward states. The approach serves as a relevant tool to study the neurons and genes that play a role in experience-mediated changes of internal state. The protocol is composed of two discrete parts: exposing the flies to rewarding and nonrewarding experiences, and assaying voluntary ethanol consumption as a measure of the motivation to obtain a drug reward. The two parts can be used independently to induce the modulation of experience as an initial step for further downstream assays or as an independent two-choice feeding assay, respectively. The protocol does not require a complicated setup and can therefore be applied in any laboratory with basic fly culture tools.

A Simple Way to Measure Alterations in Reward-seeking Behavior Using Drosophila melanogaster

We describe a protocol for inducing rewarding and nonrewarding experiences in fruit flies (Drosophila melanogaster) using voluntary ethanol consumption as a measure for changes in reward states.

Un modo semplice per misurare alterazioni in ricompensa il comportamento di ricerca Utilizzando<em&gt; Drosophila melanogaster</em

We describe a protocol for measuring ethanol self-administration in fruit flies (Drosophila melanogaster) as a proxy for changes in reward states. We ...

Measuring and Altering Mating Drive in Male Drosophila melanogaster

Despite decades of investigation, the neuronal and molecular bases of motivational states remain mysterious. We have recently developed a novel, reductionist, and scalable system for in-depth investigation of motivation using the mating drive of male Drosophila&#160;melanogaster (Drosophila), the methods for which we detail here. The behavioral paradigm centers on the finding that male mating drive decreases alongside fertility over the course of repeated copulations and recovers over ~3 d. In this system, the powerful neurogenetic tools available in the fly converge with the genetic accessibility and putative wiring diagram available for sexual behavior. This convergence allows rapid isolation and interrogation of small neuronal populations with specific motivational functions. Here we detail the design and execution of the satiety assay that is used to measure and alter courtship motivation in the male fly. Using this assay, we also demonstrate that low male mating drive can be overcome by stimulating dopaminergic neurons. The satiety assay is simple, affordable, and robust to influences of genetic background. We expect the satiety assay to generate many new insights into the neurobiology of motivational states.

Measuring and Altering Mating Drive in Male Drosophila melanogaster

This article describes a behavioral assay that uses male mating drive in Drosophila melanogaster to study motivation. Using this method, researchers can utilize advanced fly neurogenetic techniques to uncover the genetic, molecular, and cellular mechanisms that underlie this motivation.

Misurazione e l&#39;alterazione di accoppiamento Drive a Maschio<em&gt; Drosophila melanogaster</em

Despite decades of investigation, the neuronal and molecular bases of motivational states remain mysterious. We have recently developed a novel, ...

Drosophila Courtship Conditioning As a Measure of Learning and Memory

Many insights into the molecular mechanisms underlying learning and memory have been elucidated through the use of simple behavioral assays in model organisms such as the fruit fly, Drosophila melanogaster. Drosophila is useful for understanding the basic neurobiology underlying cognitive deficits resulting from mutations in genes associated with human cognitive disorders, such as intellectual disability (ID) and autism. This work describes a methodology for testing learning and memory using a classic paradigm in Drosophila known as courtship conditioning. Male flies court females using a distinct pattern of easily recognizable behaviors. Premated females are not receptive to mating and will reject the male&#39;s copulation attempts. In response to this rejection, male flies reduce their courtship behavior. This learned reduction in courtship behavior is measured over time, serving as an indicator of learning and memory. The basic numerical output of this assay is the courtship index (CI), which is defined as the percentage of time that a male spends courting during a 10 min interval. The learning index (LI) is the relative reduction of CI in flies that have been exposed to a premated female compared to na&#239;ve flies with no previous social encounters. For the statistical comparison of LIs between genotypes, a randomization test with bootstrapping is used. To illustrate how the assay can be used to address the role of a gene relating to learning and memory, the pan-neuronal knockdown of Dihydroxyacetone phosphate acyltransferase (Dhap-at) was characterized here. The human ortholog of Dhap-at, glyceronephosphate O-acyltransferase (GNPT), is involved in rhizomelic chondrodysplasia punctata type 2, an autosomal-recessive syndrome characterized by severe ID. Using the courtship conditioning assay, it was determined that Dhap-at is required for long-term memory, but not for short-term memory. This result serves as a basis for further investigation of the underlying molecular mechanisms.

Drosophila Courtship Conditioning As a Measure of Learning and Memory

This protocol describes a Drosophila learning and memory assay called courtship conditioning. This classic assay is based on a reduction of male courtship behavior after sexual rejection by a non-receptive premated female. This natural form of behavioral plasticity can be used to test learning, short-term memory, and long-term memory.

Condizionamento del corteggiamento di Drosophila come misura dell&#39;apprendimento e della memoria

Many insights into the molecular mechanisms underlying learning and memory have been elucidated through the use of simple behavioral assays in model ...

Un metodo per testare l 'effetto dei segnali ambientali sul comportamento di accoppiamento in

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Metodi per testare la disgregazione endocrina nella Drosophila melanogaster

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Un test comportamentale per Mechanosensation di cloni in marcm-based

Un modo semplice per misurare alterazioni in ricompensa il comportamento di ricerca Utilizzando

Misurazione e l'alterazione di accoppiamento Drive a Maschio

Condizionamento del corteggiamento di Drosophila come misura dell'apprendimento e della memoria