Nous tenons à remercier Astha Maltare pour générer les données crawling larvaires. Nous tenons à remercier le Dr Nicholas Lanson Jr. pour donner ses commentaires sur le manuscrit. Ce travail a été soutenu par le Centre Robert Packard pour la SLA à l&#39;Université Johns Hopkins (à l&#39;UBP) et le Sclérose latérale amyotrophique Association (UBP), et R01MH083689 de la National Institutes of Mental Health (CDN).

Essai A. Crawling larvaire 1. Collection larves <ol><li> Mettre en place une bouteille de 8 oz de mouches (10-15 mâles + femelles 10-15). </li><li> Laissez-mouches pondent leurs œufs pendant 24 heures, puis de bouteilles claire de mouches. (Transfert des adultes dans un nouveau flacon et répéter au besoin). </li><li> Incuber bouteille pendant 3-4 jours, ou jusqu&#39;au troisième stade larvaire sont visibles. </li><li> Ajouter 50 à 100 ml de saccharose à 20% à la bouteille avec les larves et les laisser reposer pendant 20 minutes. Les larves flottent à la surface. </li><li> Recueillir les larves à l&#39;aide d&#39;une pipette de 25 ml sérologique avec la pointe coupée, et le placer dans un panier en treillis. </li><li> Laver les larves dans le panier à maille deux fois avec désionisée H 2 O. Les larves sont maintenant prêts pour les expériences. </li></ol> 2. Pour traiter les larves à la drogue <ol><li> Utilisation d&#39;une brosse pour transporter le nombre souhaité de larves à un bécher contenant 5 ml de saccharose une solution à 5% + médicament. </li><li> Laissez les larves se nourrissent pendant au moins 15 minutes.</li><li> Verser la drogue traitée larves dans un panier en treillis et rincer. Ils sont maintenant prêts à utiliser. </li></ol> 3. Essai locomoteur (mesure de la distance totale parcourue ou des contractions paroi du corps) <ol><li> Utilisez une brosse pour le transport de larve à un individu: <ol><li> 15 cm boîte de Pétri contenant 2% d&#39;agarose (précédemment versé et on laisse durcir) sur du papier millimétré avec une épaisseur de 0,2 cm 2 de grille. <ol><li> Compter le nombre de lignes de la grille traversé en 1 minute. </li></ol></li><li> Eh bien d&#39;un plat de dissection verre contenant une solution de levure diluée pâte. <ol><li> Comptez le péristaltisme (contractions antérieure sur le mouvement de contraction postérieure = 1) en une minute tout en observant sous un microscope à dissection. </li></ol></li></ol></li><li> Répétez jusqu&#39;à ce que les chiffres désirés de larves ont été comptés. </li></ol> B. itératif rapide géotaxie négative (RING) Protocole Cet essai a été à l&#39;origine deprescrite par Gargano et al 5. <ol><li> Recueillir nouvellement écloses mouches mâles adultes de moins de CO 2 anesthésie légère et le placer dans un flacon standard contenant des aliments (ou de la nourriture + médicament). </li><li> Maintenir les mouches à la température ambiante (sur la paillasse. ~ 22 ° C) pendant 2-3 jours pour permettre la récupération du CO 2 (et l&#39;accumulation des concentrations de médicament l&#39;état d&#39;équilibre, le cas échéant). </li><li> Transférer environ 25 mouches sans anesthésie dans des flacons en polystyrène préparés. </li><li> Assemblez les flacons avec des mouches dans l&#39;appareil RING (Figure 1). </li><li> Permettez-mouches pour s&#39;acclimater à l&#39;environnement, à l&#39;abri, pendant 15-20 minutes. </li><li> Au cours de cet appareil photo numérique du temps lieu ~ 1 m en avant de l&#39;appareil (sur une plate-forme si nécessaire pour aligner le centre de la lentille à mi-hauteur de la flacons), mise au point et zoom de la caméra sur l&#39;appareil, et régler une minuterie pour 3,0 secondes. </li><li> Soigneusement s&#39;emparer de l&#39;appareil sonner avec votre main gauche afin de ne pas disturb les mouches, et tiennent la minuterie avec votre main droite. </li><li> Fortement appuyez sur l&#39;appareil vers le bas sur la surface du banc à trois reprises, en s&#39;assurant que l&#39;eau du robinet est déjà assez difficile de faire tomber toutes les mouches au fond des flacons. </li><li> Parallèlement à l&#39;achèvement de la troisième prise, démarrez le compte à rebours de 3 secondes. </li><li> A trois secondes de prendre une photo. </li><li> Réinitialiser la minuterie pendant 1 minute et commencer. Pendant ce temps, réinitialiser la caméra et se concentrer sur l&#39;appareil, et fixer un autre canal de la minuterie pendant trois secondes. </li><li> Après 1 minute, répétez les étapes 1.7 à 1.10 </li><li> Après un total de 5-6 essais, télécharger des images sur un ordinateur et d&#39;utiliser votre visionneuse d&#39;image préférée pour ouvrir, et marquer la hauteur moyenne a grimpé pour chaque flacon. </li><li> Effectuer une analyse statistique sur vos différents groupes en comparant la hauteur moyenne a grimpé. </li></ol> Courtship C. et dosage d&#39;accouplement <ol><li> La première chose le matin, claires et producteurs de bouteilles de flies à être utilisé. </li><li> Au cours de la journée (toutes les 3-4 heures), de recueillir nouvellement écloses mâles sexuellement naïfs et les femmes: <ol><li> Placez les hommes individuellement dans des flacons ou des tubes avec le milieu. </li><li> Placez 5-6 femelles ensemble, par flacon / tube. </li><li> Isoler les mouches recueillies à 25 ° C de moins de 12 heures de lumière / obscurité pendant 5 jours. </li><li> Transfert d&#39;une femelle dans la chambre d&#39;une roue d&#39;accouplement. </li><li> Transférer un mâle dans la chambre d&#39;une roue d&#39;accouplement. </li><li> Observez paire sous un microscope à dissection pour les comportements suivants: <ol><li> Orientation (les mâles oriente vers la femelle) </li><li> Tapping (le mâle tape la femme) </li><li> Chanson Wing (le mâle s&#39;étend et vibre d&#39;une aile) </li><li> Lèche (les mâles lèche les organes génitaux féminins) </li><li> Curling (le mâle se recourbe son abdomen sous lui-même) </li><li> Tentative de copulation (curling activité tout en essayant de monter sur la femelle) </li></ol></li></ol></li><li> Observer pour 10 minutes ou jusqu&#39;à ce que la copulation, en notant l&#39;heure à laquelle chaque problème se produit (latence), le temps total engagé dans le comportement de cour jusqu&#39;à ce que la copulation (pour calculer l&#39;indice de cour), ainsi que le nombre de paires qui ont réussi à adopter un comportement donné ( fréquence). 100% des paires de type sauvage est généralement s&#39;accoupler dans les 5 minutes. </li><li> Calculer un indice de cour (CI) en divisant le temps passé à faire la cour, divisé par le temps total jusqu&#39;à la copulation. Pour les couples de type sauvage de cette oscille habituellement entre 0.6-0.8. </li></ol> D. des résultats représentatifs Crawling test Normale larve de type sauvage va errer ~ 3 cm / minute, et d&#39;exposition de ~ 40-50 contractions paroi du corps en une minute. Nous avons récemment développé un modèle de drosophile du FUS / TLS liée sclérose latérale amyotrophique qui montre des larves rampant défaut, la durée de vie réduite et des adultes d&#39;escalade dépréciation 6. Nous avons ciblé l&#39;expression of de type sauvage et des formes mutantes de FUS / TLS afin de les neurones moteurs (OK-371-gal4 pilote) et a effectué un test exploration des larves. Comme indiqué ci-dessous, les larves de type sauvage ramper jusqu&#39;à 12 cm, alors que l&#39;expression du FUS de type sauvage a diminué la capacité des larves rampant à environ 6 cm. Animaux exprimant la mutation R521C SLA causant dans FUS / TLS démontrer une déficience très sévère dans leur mouvement rampant (Figure 1), ramper seulement d&#39;environ 1 cm par minute. Négatif dosage RING géotaxie Les jeunes mouches adultes de type sauvage doit avoir une hauteur moyenne d&#39;escalade de ~ 4-5 cm dans un délai de 3 secondes (le temps peut être réglé de 3 secondes pour accueillir différentes souches ou des niveaux d&#39;activité pour définir une hauteur moyenne donnée pour une donnée la souche / traitement). Mouches qui restent à la partie inférieure est attribuée une valeur de 0. Il n&#39;est pas conseillé d&#39;utiliser plus de 25 mouches par flacon, car il devient alors difficile de déterminer l&#39;emplacement de chaque individu à la mouche pour mesurer la hauteur. Ndésensibilisation o a été observée à la hauteur des 6 essais consécutifs espacés de 1 minute d&#39;intervalle que nous avons employée. Il est essentiel de ne pas réutiliser les flacons d&#39;essai en polystyrène dans cet essai, après les premières poses de données sont recueillies en raison de nouvelles mouches placées dans des flacons utilisés ne veut pas monter dans la même mesure que dans des flacons frais. <img alt="Figure 1" src="/files/ftp_upload/3795/3795fig1.jpg" /> Figure 1. Configuration pour le test RING. L&#39;appareil photo numérique est placé ~ 1 m en avant de l&#39;appareil contenant les mouches dans les tubes en polystyrène; mise au point et zoom de la caméra sur l&#39;appareil et régler une minuterie pour 3,0 secondes. <img alt="Figure 2" src="/files/ftp_upload/3795/3795fig2.jpg" /> Les données représentatives Figure 2. De l&#39;essai ramper larves à l&#39;aide mouches ectopique exprimer UAS-FUS WT, et UAS-FUS R521C sous le contrôle d&#39;un conducteur du neurone moteur (OK371-Gal4). <pclass = &quot;jove_content&quot;&gt;

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Nous n&#39;avons rien à communiquer.

Comportement chez la drosophile est étroitement régulée par des facteurs génétiques et environnementaux. Nous, et d&#39;autres, ont déjà utilisé les tests décrits ici pour recueillir des données afin d&#39;examiner les gènes liés à voler et à des comportements humains les maladies neurodégénératives modélisés chez la drosophile 5-19. Pour le dosage de l&#39;exploration, la sélection minutieuse de 3 e stade larvaire est une étape critique. Si le traitement avec...

<table border="1"><tbody><tr><td> Nom du réactif </td><td> Entreprise </td><td> Numéro de catalogue </td></tr><tr><td> Saccharose </td><td> Fisher Scientific </td><td> S5-500 </td></tr><tr><td> Agarose </td><td> Invitrogen </td><td> 16500-500 </td></tr><tr><td> 6 oz Drosophila bouteille </td><td> Genesee scientifique </td><td> 32-130 </td></tr><tr><td> Paint Brush (# 1) </td><td> Ted Pella, Inc </td><td> 11859 </td></tr><tr><td> Fly composants alimentaires </td><td></td><td></td></tr><tr><td> Semoule de maïs </td><td> Fisher Scientific </td><td> NC9109741 </td></tr><tr><td> Gélose </td><td> Genesee scientifique </td><td> 66-104 </td></tr><tr><td> Mélasse </td><td> Fisher Scientific </td><td> NC9349176 </td></tr><tr><td> L&#39;acide propionique </td><td> Acros </td><td> 14930-0010 </td></tr><tr><td> Tegosept </td><td> Apex </td><td> 20-258 </td></tr><tr><td> L&#39;éthanol </td><td> PêcheurScientifique </td><td> BP2818-4 </td></tr><tr><td> Levure </td><td> Genesee scientifique </td><td> 62-107 </td></tr></tbody></table>

methods to assay drosophila behavior

Drosophila melanogaster, la mouche des fruits, a été utilisé pour étudier les mécanismes moléculaires d&#39;un large éventail de maladies humaines comme le cancer, les maladies cardiovasculaires et de diverses maladies neurologiques 1. Nous avons optimisé simples et robustes tests comportementaux pour déterminer locomotion de la larve, la capacité d&#39;escalade adulte (dosage RING), et les comportements de séduction de la drosophile. Ces tests comportementaux sont largement applicables pour étudier le rôle des facteurs génétiques et environnementaux sur le comportement à la mouche. La capacité de ramper larvaire peut être utilisé de manière fiable pour déterminer les changements à un stade précoce dans les capacités rampants de larves de drosophile et aussi pour examiner l&#39;effet de médicaments ou de gènes de maladies humaines (des mouches transgéniques) sur leur locomotion. Le dosage des larves rampant devient plus applicable si l&#39;expression ou la suppression d&#39;un gène provoque une létalité à un stade nymphal ou adulte, que ces mouches ne survivent pas à l&#39;âge adulte où ils pourraient par ailleurs être évalué. Cette base, unssay peut également être utilisé en conjonction avec la lumière vive ou le stress d&#39;examiner d&#39;autres réponses comportementales chez les larves de Drosophila. Comportement de cour a été largement utilisée pour étudier les bases génétiques du comportement sexuel, et peut également être utilisé pour examiner l&#39;activité et de la coordination, ainsi que l&#39;apprentissage et la mémoire. Comportement de cour chez la drosophile implique l&#39;échange de divers stimuli sensoriels, y compris visuelles, auditives, et les signaux chimiosensoriels entre les hommes et les femmes qui mènent à une série complexe de comportements moteurs bien caractérisés ont abouti à la copulation avec succès. Dosages adultes traditionnels d&#39;escalade (géotaxie négatifs) sont fastidieux, de main-d&#39;œuvre, et prend du temps, avec une variation significative entre les différents essais 2-4. Le itératif rapide géotaxie négative (RING) test 5 a de nombreux avantages sur des protocoles plus largement utilisés, en fournissant une approche débit reproductible, sensible, et de haute de quantifier locomotrice des adultes et négatifs géotaxie behaviors. Dans le test RING, plusieurs génotypes ou de traitements médicamenteux peuvent être testés simultanément en utilisant grand nombre d&#39;animaux, à l&#39;approche à haut débit rend plus propice pour des expériences de dépistage.

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Methods to Assay Drosophila Behavior

Drosophila melanogaster est un système modèle génétiquement et comportemental souple qui a été utilisée pour comprendre la base moléculaire et cellulaire de nombreux processus biologiques importants pour plus d&#39;un siècle 1. Drosophile a été bien exploitée afin de mieux comprendre la base génétique du comportement à la mouche.

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 ...

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Neuroscience

Methods to Assay Drosophila Behavior

42.7K vues.  Louisiana State University Health Sciences Center. Drosophila melanogaster est un système modèle génétiquement et comportemental souple qui a été utilisée pour comprendre la base moléculaire et cellulaire de nombreux processus biologiques importants pour plus d'un siècle 1. Drosophile a été bien exploitée afin de mieux comprendre la base génétique du comportement à la mouche.

Vidéo: Methods to Assay Drosophila Behavior

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

The sense of smell is essential for insects to find foods, mates, predators, and oviposition sites3. Insect olfactory sensory neurons (OSNs) are enclosed in sensory hairs called sensilla, which cover the surface of olfactory organs. The surface of each sensillum is covered with tiny pores, through which odorants pass and dissolve in a fluid called sensillum lymph, which bathes the sensory dendrites of the OSNs housed in a given sensillum. The OSN dendrites express odorant receptor (OR) proteins, which in insects function as odor-gated ion channels4, 5. The interaction of odorants with ORs either increases or decreases the basal firing rate of the OSN. This neuronal activity in the form of action potentials embodies the first representation of the quality, intensity, and temporal characteristics of the odorant6, 7.
Given the easy access to these sensory hairs, it is possible to perform extracellular recordings from single OSNs by introducing a recording electrode into the sensillum lymph, while the reference electrode is placed in the lymph of the eye or body of the insect. In Drosophila, sensilla house between one and four OSNs, but each OSN typically displays a characteristic spike amplitude. Spike sorting techniques make it possible to assign spiking responses to individual OSNs. This single sensillum recording (SSR) technique monitors the difference in potential between the sensillum lymph and the reference electrode as electrical spikes that are generated by the receptor activity on OSNs1, 2, 8. Changes in the number of spikes in response to the odorant represent the cellular basis of odor coding in insects. Here, we describe the preparation method currently used in our lab to perform SSR on Drosophila melanogaster and Anopheles gambiae, and show representative traces induced by the odorants in a sensillum-specific manner.

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

Electrophysiological responses of olfactory sensory neurons to odorants can be measured in insects using single sensillum recordings. In this video article we will demonstrate how to perform single sensillum recordings in the antennae of the vinegar fly (Drosophila melanogaster) and the maxillary palps of the malaria mosquito (Anopheles gambiae).

Enregistrements Sensillum simple chez les insectes Drosophila melanogaster Et Anopheles gambiae</em

The sense of smell is essential for insects to find foods, mates, predators, and oviposition sites3. Insect olfactory sensory neurons (OSNs) are enclosed ...

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Neurosciences

Methods for Study of Neuronal Morphogenesis: Ex vivo RNAi Electroporation in Embryonic Murine Cerebral Cortex

The cerebral cortex directs higher cognitive functions. This six layered structure is generated in an inside-first, outside-last manner, in which the first born neurons remain closer to the ventricle while the last born neurons migrate past the first born neurons towards the surface of the brain1. In addition to neuronal migration2, a key process for normal cortical function is the regulation of neuronal morphogenesis3. While neuronal morphogenesis can be studied in vitro in primary cultures, there is much to be learned from how these processes are regulated in tissue environments.
We describe techniques to analyze neuronal migration and/or morphogenesis in organotypic slices of the cerebral cortex4,6. A pSilencer modified vector is used which contains both a U6 promoter that drives the double stranded hairpin RNA and a separate expression cassette that encodes GFP protein driven by a CMV promoter7-9. Our approach allows for the rapid assessment of defects in neurite outgrowth upon specific knockdown of candidate genes and has been successfully used in a screen for regulators of neurite outgrowth8. Because only a subset of cells will express the RNAi constructs, the organotypic slices allow for a mosaic analysis of the potential phenotypes. Moreover, because this analysis is done in a near approximation of the in vivo environment, it provides a low cost and rapid alternative to the generation of transgenic or knockout animals for genes of unknown cortical function. Finally, in comparison with in vivo electroporation technology, the success of ex vivo electroporation experiments is not dependant upon proficient surgery skill development and can be performed with a shorter training time and skill.

Methods for Study of Neuronal Morphogenesis: Ex vivo RNAi Electroporation in Embryonic Murine Cerebral Cortex

To conduct a rapid assessment of the function of genes in the development of cerebral cortex, we describe methods involving the ex vivo electroporation of plasmids co-expressing inhibitory RNA (RNAi) and GFP in murine embryonic cortex. This protocol is amenable to the study of various aspects of neurodevelopment such as neurogenesis, neuronal migration and neuronal morphogenesis including dendrite and axon outgrowth.

Méthodes pour l&#39;étude de la morphogenèse neuronale:<em&gt; Ex vivo</em&gt; Électroporation ARNi dans embryonnaire cortex cérébral murin

The cerebral cortex directs higher cognitive functions. This six layered structure is generated in an inside-first, outside-last manner, in which the ...

Local and Global Methods of Assessing Thermal Nociception in Drosophila Larvae

In this article, we demonstrate assays to study thermal nociception in Drosophila larvae. One assay involves spatially-restricted (local) stimulation of thermal nociceptors1,2 while the second involves a wholesale (global) activation of most or all such neurons3. Together, these techniques allow visualization and quantification of the behavioral functions of Drosophila nociceptive sensory neurons.
The Drosophila larva is an established model system to study thermal nociception, a sensory response to potentially harmful temperatures that is evolutionarily conserved across species1,2. The advantages of Drosophila for such studies are the relative simplicity of its nervous system and the sophistication of the genetic techniques that can be used to dissect the molecular basis of the underlying biology4-6 In Drosophila, as in all metazoans, the response to noxious thermal stimuli generally involves a "nocifensive" aversive withdrawal to the presented stimulus7. Such stimuli are detected through free nerve endings or nociceptors and the amplitude of the organismal response depends on the number of nociceptors receiving the noxious stimulus8. In Drosophila, it is the class IV dendritic arborization sensory neurons that detect noxious thermal and mechanical stimuli9 in addition to their recently discovered role as photoreceptors10. These neurons, which have been very well studied at the developmental level, arborize over the barrier epidermal sheet and make contacts with nearly all epidermal cells11,12. The single axon of each class IV neuron projects into the ventral nerve cord of the central nervous system11 where they may connect to second-order neurons that project to the brain.
Under baseline conditions, nociceptive sensory neurons will not fire until a relatively high threshold is reached. The assays described here allow the investigator to quantify baseline behavioral responses or, presumably, the sensitization that ensues following tissue damage. Each assay provokes distinct but related locomotory behavioral responses to noxious thermal stimuli and permits the researcher to visualize and quantify various aspects of thermal nociception in Drosophila larvae. The assays can be applied to larvae of desired genotypes or to larvae raised under different environmental conditions that might impact nociception. Since thermal nociception is conserved across species, the findings gleaned from genetic dissection in Drosophila will likely inform our understanding of thermal nociception in other species, including vertebrates.

Local and Global Methods of Assessing Thermal Nociception in Drosophila Larvae

In this article, we demonstrate assays to study thermal nociception in Drosophila larvae. One assay involves spatially-restricted (local) stimulation of thermal nociceptors1,2 while the second involves a wholesale (global) activation of most or all such neurons3. Together, these techniques allow visualization and quantification of the behavioral functions of Drosophila nociceptive sensory neurons.

Méthodes locales et globales de l&#39;évaluation de nociception thermique dans<em&gt; Drosophila</em&gt; Les larves

In this article, we demonstrate assays to study thermal nociception in Drosophila larvae. One assay involves spatially-restricted (local) stimulation of ...

Optogenetic Stimulation of Escape Behavior in Drosophila melanogaster

A growing number of genetically encoded tools are becoming available that allow non-invasive manipulation of the neural activity of specific neurons in Drosophila melanogaster1. Chief among these are optogenetic tools, which enable the activation or silencing of specific neurons in the intact and freely moving animal using bright light. Channelrhodopsin (ChR2) is a light-activated cation channel that, when activated by blue light, causes depolarization of neurons that express it. ChR2 has been effective for identifying neurons critical for specific behaviors, such as CO2 avoidance, proboscis extension and giant-fiber mediated startle response2-4. However, as the intense light sources used to stimulate ChR2 also stimulate photoreceptors, these optogenetic techniques have not previously been used in the visual system. Here, we combine an optogenetic approach with a mutation that impairs phototransduction to demonstrate that activation of a cluster of loom-sensitive neurons in the fly's optic lobe, Foma-1 neurons, can drive an escape behavior used to avoid collision. We used a null allele of a critical component of the phototransduction cascade, phospholipase C-&beta;, encoded by the norpA gene, to render the flies blind and also use the Gal4-UAS transcriptional activator system to drive expression of ChR2 in the Foma-1 neurons. Individual flies are placed on a small platform surrounded by blue LEDs. When the LEDs are illuminated, the flies quickly take-off into flight, in a manner similar to visually driven loom-escape behavior. We believe that this technique can be easily adapted to examine other behaviors in freely moving flies.

Optogenetic Stimulation of Escape Behavior in Drosophila melanogaster

Genetically encoded optogenetic tools enable noninvasive manipulation of specific neurons in the Drosophila brain. Such tools can identify neurons whose activation is sufficient to elicit or suppress particular behaviors. Here we present a method for activating Channelrhodopsin2 that is expressed in targeted neurons in freely walking flies.

Stimulation optogenetic de comportement de fuite en<em&gt; Drosophila melanogaster</em

A  growing number of genetically encoded tools are becoming available that allow non-invasive  manipulation of the neural activity of specific neurons in ...

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 ...

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 ...

Using Linear Agarose Channels to Study Drosophila Larval Crawling Behavior

Drosophila larval crawling is emerging as a powerful model to study neural control of sensorimotor behavior. However, larval crawling behavior on flat open surfaces is complex,&#160;including: pausing, turning, and meandering. This complexity in the repertoire of movement hinders detailed analysis of the events occurring during a single crawl stride cycle. To overcome this obstacle, linear agarose channels were made that constrain larval behavior to straight, sustained, rhythmic crawling. In principle, because agarose channels and the Drosophila larval body are both optically clear, the movement of larval structures labeled by genetically-encoded fluorescent probes can be monitored in intact, freely-moving larvae. In the past, larvae were placed in linear channels and crawling at the level of whole organism, segment, and muscle were analyzed1. In the future, larvae crawling in channels can be used for calcium imaging to monitor neuronal activity. Moreover, these methods can be used with larvae of any genotype and with any researcher-designed channel. Thus the protocol presented below is widely applicable for studies using the Drosophila larva as a model to understand motor control.

Using Linear Agarose Channels to Study Drosophila Larval Crawling Behavior

The Drosophila larva is a powerful model system to study neural control of behavior. This publication describes the use of linear agarose channels to elicit sustained bouts of linear crawling and methods to quantify the dynamics of larval structures during repetitive crawling behavior.

Utilisation Agarose linéaire Canaux pour étudier<em&gt; Drosophila</em&gt; Larvaire Ramper Comportement

Drosophila larval crawling is emerging as a powerful model to study neural control of sensorimotor behavior. However, larval crawling behavior on flat ...

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 ...

Quantification of Drosophila Grooming Behavior

Drosophila grooming behavior is a complex multi-step locomotor program that requires coordinated movement of both forelegs and hindlegs. Here we present a grooming assay protocol and novel chamber design that is cost-efficient and scalable for either small or large-scale studies of Drosophila grooming. Flies are dusted all over their body with Brilliant Yellow dye and given time to remove the dye from their bodies within the chamber. Flies are then deposited in a set volume of ethanol to solubilize the dye. The relative spectral absorbance of dye-ethanol samples for groomed versus ungroomed animals are measured and recorded. The protocol yields quantitative data of dye accumulation for individual flies, which can be easily averaged and compared across samples. This allows experimental designs to easily evaluate grooming ability for mutant animal studies or circuit manipulations. This efficient procedure is both versatile and scalable. We show work-flow of the protocol and comparative data between WT animals and mutant animals for the Drosophila type I Dopamine Receptor (DopR).

This protocol describes a scalable individual grooming assay technique in Drosophila that yields robust, quantitative data to measure grooming behavior. The method is based on comparing the difference in dye accumulation on the bodies of un-groomed versus groomed animals over a set period of time.

Quantification du comportement de toilettage de la drosophile

Drosophila grooming behavior is a complex multi-step locomotor program that requires coordinated movement of both forelegs and hindlegs. Here we present a ...

High-throughput Analysis of Locomotor Behavior in the Drosophila Island Assay

Advances in next-generation sequencing technologies contribute to the identification of (candidate) disease genes for movement disorders and other neurological diseases at an increasing speed. However, little is known about the molecular mechanisms that underlie these disorders. The genetic, molecular, and behavioral toolbox of Drosophila melanogaster makes this model organism particularly useful to characterize new disease genes and mechanisms in a high-throughput manner. Nevertheless, high-throughput screens require efficient and reliable assays that, ideally, are cost-effective and allow for the automatized quantification of traits relevant to these disorders. The island assay is a cost-effective and easily set-up method to evaluate Drosophila locomotor behavior. In this assay, flies are thrown onto a platform from a fixed height. This induces an innate motor response that enables the flies to escape from the platform within seconds. At present, quantitative analyses of filmed island assays are done manually, which is a laborious undertaking, particularly when performing large screens.
This manuscript describes the &#34;Drosophila Island Assay&#34; and &#34;Island Assay Analysis&#34; algorithms for&#160;high-throughput, automated data processing and quantification of island assay data. In the setup, a simple webcam connected to a laptop collects an image series of the platform while the assay is performed. The &#34;Drosophila Island Assay&#34; algorithm developed for the open-source software Fiji processes these image series and quantifies, for each experimental condition, the number of flies on the platform over time. The &#34;Island Assay Analysis&#34; script, compatible with the free software R, was developed to automatically process the obtained data and to calculate whether treatments/genotypes are statistically different. This greatly improves the efficiency of the island assay and makes it a powerful readout for basic locomotion and flight behavior. It can thus be applied to large screens investigating fly locomotor ability, Drosophila models of movement disorders, and drug efficacy.

High-throughput Analysis of Locomotor Behavior in the Drosophila Island Assay

The island assay is a relatively new, cost-effective assay that can be used to evaluate the basic locomotor behavior of Drosophila melanogaster. This manuscript describes algorithms for automatic data processing and objective quantification of island assay data, making this assay a sensitive, high-throughput readout for large genetic or pharmacological screens.