Nous remercions le Bloomington Drosophila Stock Center pour les stocks de mouches; C. Gahr, JT Alkema et S. van Hasselt pour leur tentative initiale de développer le dosage de l&#39;air sous pression; Jasper Bosman pour les conseils sur la culture de la levure; Et Rezza Azanchi et Joel Levine pour l&#39;origine du développement de la surveillance de la durée du comportement d&#39;accouplement de Drosophila . JA Gorter a été soutenu par une bourse de programme d&#39;études supérieures BCN / NWO de recherche en neurosciences. Ce travail a été soutenu en partie par l&#39;organisation néerlandaise pour la recherche scientifique (NWO) (référence: 821.02.020) à JC Billeter.

<ol>
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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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Les auteurs n&#39;ont aucun intérêt concurrentiel à divulguer.

Ce protocole décrit un dosage pour tester le comportement d&#39;accouplement sur 24 h tout en contrôlant en permanence les indices environnementaux qu&#39;un couple d&#39;accouplement a l&#39;hypothèse d&#39;utiliser pour déterminer la fréquence d&#39;accouplement. Il est possible d&#39;augmenter la fréquence d&#39;accouplement en réponse à l&#39;air de levure délivré par un système d&#39;air sous pression lorsque le milieu contient également de la levure ( figure 2B ). En outre, une répons...

Les comportements reproductifs ont généralement des coûts énergétiques élevés, en particulier pour les femmes, qui produisent des gamètes plus grands que les mâles et doivent soigneusement choisir les conditions pour élever leur progéniture en développement. En raison du coût de l&#39;énergie, il n&#39;est pas surprenant que la reproduction soit liée aux conditions nutritionnelles. Cela est vrai dans la plupart des animaux, y compris les animaux, y compris les mammifères, dont la puberté peut être retardée par la malnutrition et dont le comportement sexuel peut être affecté négativement par la restriction alimentaire 1 . La reproduction de l&#39;organisme modèle génétique Drosophila melanogaster est également affectée par des conditions nutritionnelles. Le tribunal masculin à un niveau supérieur en présence de substances volatiles alimentaires 2 , et les femelles sont plus sexuellement réceptives en présence de levure, un nutriment majeur pour la production d&#39;œufs et la survie de la progéniture 3 , 4 , 5 . CeLa réponse reproductive reproductive évolutive aux aliments offre la possibilité d&#39;étudier des mécanismes qui relient la disponibilité alimentaire environnementale à la reproduction sexuelle dans un organisme génétiquement traitable et efficace dans le temps. En effet, le travail dans D. melanogaster a impliqué la voie de l&#39;insuline comme un régulateur important de la connexion entre la nourriture et le comportement d&#39;accouplement 6 . Il a également montré que l&#39;acte d&#39;accouplement change la préférence alimentaire des femelles ainsi que les neurones chimiosensibles associés 7 , 8 , 9 . Il est clair que les indices alimentaires affectent les comportements reproductifs chez D. melanogaster . Ces effets semblent affecter principalement les femmes, en particulier celles qui ont déjà accouplé 5 . Cependant, pour tester ces effets aigus des conditions environnementales, le dosage classiquement utilisé pour le comportement d&#39;accouplement féminin pourraitNe pas être très approprié en raison des longues interruptions entre les épisodes d&#39;accouplement. Dans l&#39;essai de remix classique, une femelle vierge s&#39;accouple avec un homme et est immédiatement isolée et présente un nouveau mâle de 24 à 48 h plus tard. Cet essai classique a été utilisé avec beaucoup de succès pour identifier les composants de l&#39;éjaculation masculin qui modifient le comportement féminin et la réponse féminine 12 , 13 , 14 , 15 , 16 , 17 , 18 . L&#39;essai d&#39;accouplement continu démontré ici, est donc un ajout aux essais d&#39;accouplement classiques qui peuvent être utilisés pour étudier l&#39;effet aigu des conditions environnementales sur les comportements reproductifs. En utilisant l&#39;analyse continue du comportement d&#39;accouplement qui est expliqué ici, nous avons précédemment montré qu&#39;une paire de mouches exposées à la levure remettait sChaque fois sur une période d&#39;observation de 24 h 5 , 19 , 20 , 21 , alors que les mouches ne sont pas exposées à la nourriture ne seront remises qu&#39;une fois 5 . Cette découverte peut être troublante à la lumière d&#39;une grande partie de la littérature de D. melanogaster indiquant que les femelles ne se remèment pas pendant plusieurs jours après un accouplement initial (examiné dans les références 10 , 11 ). Cependant, cette divergence s&#39;explique facilement par des conditions de dosage, où une femme est isolée pendant un à plusieurs jours avant qu&#39;une nouvelle opportunité d&#39;accouplement ne soit fournie. Si la paire ne s&#39;accroche pas dans cette période d&#39;observation d&#39;une heure, la femelle est caractérisée comme non réceptive. En outre, la fréquence d&#39;accouplement élevée ne devrait pas être surprenante étant donné que les données provenant de mouches sauvages montrent que les femelles contiennent des spermatozoïdes de 4 à 6 mâles dans leurs organes de stockage; Donc dansDisant que les femelles se remètent naturellement à plusieurs reprises 22 , 23 . Ici, nous démontrons l&#39;utilisation de cet essai d&#39;accouplement continu pour décrire comment les mouches se rassemblent et combinent des informations sur les conditions environnementales pour moduler la fréquence d&#39;accouplement. Ce test permet de tester un nombre relativement élevé de couples d&#39;accouplement pour des études génétiques et de tester l&#39;influence de signaux environnementaux volatiles et non volatiles. Le dosage fonctionne généralement pendant 24 h, mais peut être prolongé jusqu&#39;à 48 h, ce qui permet de tester les signaux environnementaux cycliques tels que le cycle lumière-sombre (LD). Nous démontrons ce dosage en testant l&#39;influence des indices volatiles d&#39;une culture de levure dans un système d&#39;air sous pression en combinaison avec la disponibilité de nutriments de levure non volatiles dans le substrat alimentaire. Le système d&#39;air sous pression pompe continuellement des signaux volatiles dans une arène d&#39;accouplement qui contientUn substrat alimentaire et un couple de test (dont le comportement d&#39;accouplement est surveillé). Pour déterminer davantage les spécificités par lesquelles la levure influence l&#39;accouplement, nous testons un composé volatil majeur de levure, à savoir l&#39;acide acétique 24 , en combinaison avec une teneur en acides aminés qui correspond à celle de la levure dans le substrat alimentaire, sous forme de peptone (amino Acides dérivés de la digestion enzymatique des protéines animales). Ensemble, ces expériences démontrent comment l&#39;effet des indices environnementaux sur le comportement d&#39;accouplement de D. melanogaster peut être testé avec ce dosage.

<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 (pure)</td><td>BD</td><td>214530</td><td>bacto</td></tr><tr><td>Glucose (0(+)-glucose monohydrate)&amp;nbsp;</td><td>Merck</td><td>18270000004</td><td></td></tr><tr><td>Open caps</td><td>Schott</td><td>29 240 28&amp;nbsp;</td><td>GL45</td></tr><tr><td>Silicone septum</td><td>VWR</td><td>548-0662</td><td></td></tr><tr><td>Barbed bulkhead fittings</td><td>Nalgene</td><td>6149-0002</td><td></td></tr><tr><td>Large PVC tubing</td><td></td><td></td><td>diameter: outer 1.2 cm and inner 0.9 cm</td></tr><tr><td>Small PVC tubing</td><td></td><td></td><td>diameters: outer 0.8 cm and inner 0.5 cm</td></tr><tr><td>15 ml tube</td><td>Falcon</td><td></td><td></td></tr><tr><td>Aquarium pump</td><td>Sera precision</td><td></td><td>Sera air 110 plus, AC 220-240 V, 50/60 Hz, 3 W and pressure &gt;100 mbar</td></tr><tr><td>Activated charcoal</td><td>Superfish</td><td>A8040400</td><td>Norit activated carbon</td></tr><tr><td>Disposible filter 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

Le comportement sexuel d&#39;un individu est influencé par le génotype, l&#39;expérience et les conditions environnementales. La façon dont ces facteurs interagissent pour moduler les comportements sexuels reste mal comprise. Dans Drosophila melanogaster , les indices environnementaux, tels que la disponibilité alimentaire, affectent l&#39;activité d&#39;accouplement, offrant un système de traçabilité permettant d&#39;étudier les mécanismes qui modulent les comportements sexuels. Dans D. melanogaster , les signaux environnementaux sont souvent détectés via les systèmes gustatifs chemosensoriels et olfactifs. Ici, nous présentons une méthode pour tester l&#39;effet des indices chimiques environnementaux sur le comportement d&#39;accouplement. Le dosage consiste en une petite arène d&#39;accouplement contenant du milieu alimentaire et un couple d&#39;accouplement. La fréquence d&#39;accouplement pour chaque couple est surveillée en permanence pendant 24 h. Nous présentons ici l&#39;applicabilité de ce dosage pour tester les composés environnementaux à partir d&#39;une source externe à travers un système d&#39;air sous pression ainsi que la manipulation des composants environnementaux directement dans l&#39;arène d&#39;accouplement. VousLa présence d&#39;un système d&#39;air sous pression est particulièrement utile pour tester l&#39;effet de composés très volatils, tout en manipulant des composants directement dans l&#39;arène d&#39;accouplement peut être utile pour déterminer la présence d&#39;un composé. Ce dosage peut être adapté pour répondre aux questions sur l&#39;influence des indices génétiques et environnementaux sur le comportement d&#39;accouplement et la fécondité ainsi que sur d&#39;autres comportements reproducteurs masculins et féminins.

L&#39;utilisation de cet essai continu, le comportement d&#39;accouplement et la fréquence d&#39;accouplement en particulier peuvent être déterminés dans des conditions environnementales expérimentales. Pour contrôler les conditions environnementales, nous avons transformé une armoire de cuisine en acier inoxydable en une zone d&#39;essai, avec sa propre source de lumière et sa diffusion, ce qui garantit une grande quantité de lumière et une quantité minimale d&#39;éblouissem...

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

Nous démontrons un dosage pour analyser les indices environnementaux et génétiques qui influent sur le comportement d&#39;accouplement dans la mouche des fruits Drosophila melanogaster .

Une méthode pour tester l&#39;effet des cotes environnementales sur le comportement d&#39;accouplement dans<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 ...

a-method-to-test-effect-environmental-cues-on-mating-behavior

Universidad de Cartagena UDEC

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Behavior

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

9.0K vues.  University of Groningen.  Nous démontrons un dosage pour analyser les indices environnementaux et génétiques qui influent sur le comportement d'accouplement dans la mouche des fruits Drosophila melanogaster . 

Vidéo: 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.

Méthodes de dosage à Drosophila Comportement

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

Recherche

Neurosciences

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.

Une simple mouche Essai de comportement alimentaire dans<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 ...

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.

Essai simple pour la quantification de l&#39;évitement social en<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.

Insectes teinture pour comportement essais: le comportement d&#39;accouplement des anesthésié<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.

Conditions affectant espace social<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 New Approach that Eliminates Handling for Studying Aggression and the "Loser" Effect in Drosophila melanogaster

Aggressive behavior in Drosophila melanogaster is composed of the sequential expression of stereotypical behavioral patterns (for analysis see 1). This complex behavior is influenced by genetic, hormonal and environmental factors. As in many organisms, previous fighting experience influences the fighting strategy of flies and the outcome of later contests: losing a fight increases the probability of losing later contests, revealing &#34;loser&#34; effects that likely involve learning and memory 2-4. The learning and memory that accompanies expression of complex social behaviors like aggression, is sensitive to pre-test handling of animals 5,6. Many experimental procedures are used in different laboratories to study aggression 7-9, however, no routinely used protocol that excludes handling of flies is currently available. Here, we report a new behavioral apparatus that eliminates handling of flies, using instead their innate negative geotactic responses to move animals into or out of fighting chambers. In this protocol, small circular fight arenas containing a food cup are divided into two equal halves by a removable plastic slider prior to introduction of flies. Flies enter chambers from their home isolation vials via sliding chamber doors and geotaxis. Upon removal of plastic sliders, flies are free to interact. After specified time periods, flies are separated again by sliders for subsequent experimentation. All of this is done easily without handling of individual flies. This apparatus offers a novel approach to study aggression and the associated learning and memory, including the formation of &#34;loser&#34; effects in fly fights. In addition, this new general-purpose behavioral apparatus can be employed to study other social behaviors of flies and should, in general, be of interest for investigating experience-related changes in fundamental behavioral processes.

A New Approach that Eliminates Handling for Studying Aggression and the "Loser" Effect in Drosophila melanogaster

During fruit fly fights, the behavioral patterns observed, fight dynamics, and associated learning and memory are influenced by experimental conditions. The protocol presented here describes a novel procedure that entirely eliminates handling of flies during experiments. This improves fight dynamics and allows formation of strong &#34;loser&#34; effects.

Une nouvelle approche qui élimine la manipulation pour l&#39;étude de l&#39;agression et le «perdant» dans Effet<em&gt; Drosophila melanogaster</em

Aggressive behavior in Drosophila melanogaster is composed of the sequential expression of stereotypical behavioral patterns (for analysis see 1). This ...

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.

A Behavioral Essai pour mécanosensibilité de marcm-fondé clones dans<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 moyen simple pour mesurer Transformations en récompense le comportement de recherche Utilisation<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.

Mesure et Altération Mating Drive à Male<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.

Drosophila Courtship Conditionnement en tant que mesure de l&#39;apprentissage et de la mémoire

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

Une méthode pour tester l'effet des cotes environnementales sur le comportement d'accouplement dans

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