Wir danken 4 anonymen Gutachtern für ihre Arbeit, das Manuskript zu verbessern. Wir danken dem Centre National de la Recherche Scientifique für die finanzielle Unterstützung, um MBG und YG, und der Université de Bourgogne und der Französisch Ministerium für Forschung, um MMS. Research in YG Labor wird von der Europäischen Forschungsrates (ERC Starting Grant, GliSFCo-311403), der Agence Nationale de la Recherche (ANR-JCJC, GGCB-2010), des Conseil Régional de Bourgogne (Faber) und dem CNRS finanziert.

1. Vor dem Start <ol><li> Verwenden Sie ein isogenized Referenz Lager Lager stabil und robust Verhaltens Phänotypen. Es gibt keine allgemeine Regel für die Wahl dieses Lager, da alle potenziellen Kontrollen können heterogenen Hintergrund Allele tragen. </li><li> Verwenden Sie dieses Kontrollstamm, jede andere Lager, die für spätere Schritte Rückkreuzung. Diese Rückkreuzung Schritt wird in der Regel um mindestens 5 aufeinander folgenden Kreuzungen eines einzigen weiblichen Jungfrau vertreten zu 2-3 isogene Vergleichs Männer 5 (mögliche Crossing-over zwischen homologen Chromosomen zu ermöglichen). Dieser Schritt ist wichtig, um den genetischen Hintergrund der verschiedenen Flugbestände homogenisieren. </li><li> Pflegen Drosophila Aktien auf einem Standard-Maismehl (9%), Hefe (10%) und Agar-Medium (1,5%) mit einer Antibiotika-(0,4% Methyl-para-Hydroxy-Benzoat) in einem 12-Stunden Licht / Dunkel-Zyklus ergänzt bei 25 ° C. </li><li> Erreichen Sie chemosensory Experimente in einem temperierten Raum (25 ° C) unter viel rotes Licht(Um den Beitrag der visuellen Hinweisen zu beseitigen und die auf chemosensorischen Signale konzentrieren). Regelmäßig erneuern die Luft des Raumes, um den Bereich zwischen jedem Versuch zu lüften. </li></ol> 2. Olfaktorischen Reaktion mit einem Y-Labyrinth-Test <ol><li> Verhungern die Fliegen für 16-18 h bei 25 ° C in Glasröhrchen, die nassen Papiertuch vor der Prüfung. </li><li> Nehmen Sie an einer Y-Form-Stecker an zwei Glasflaschen und zu einem kleineren Kunststofffläschchen (Lade Fläschchen). Verwenden Sie 1 ml Pipettenspitzen, die durch die Schaumstopper geben, um den Anschluss an die drei Fläschchen zu verknüpfen und einen dicht verschlossenen Y-Labyrinth zu erhalten. Schneiden Sie die schmalen Enden von zwei Pipettenspitzen (~ 2 mm Durchmesser, um eine Rückkehr der Fliege zu vermeiden, sobald er seine Entscheidung gemacht), um zwei &quot;Falle&quot; Fläschchen zu bilden, und ein großes Ende einer Pipettenspitze, um den &quot;Laden&quot; zu bilden Rohr (1A). </li><li> Kurz vor dem Anschluss der &quot;Falle&quot; Fläschchen (1B), legen Sie ein ~ 6 mm Durchmesser Filterpapier injedes Fläschchen. 40 &amp;mgr; l Geruchsstoff-Lösung auf ein Filterpapier, und 40 ul des entsprechenden Lösungsmittels auf dem zweiten Filterpapier. </li><li> Führen zehn 4-9 Tage alten Fliegen in der &quot;loading&quot; Fläschchen. Verwenden Sie keine CO 2-Narkose während dieser Überweisung, da es einen starken Einfluss auf das Verhalten der 10 hat. Vielmehr verwenden kurzem Abkühlen auf Eis. Die richtige Handhabung der narkotisierten Fliegen ist wichtig, um Stress zu den Themen so weit wie möglich zu begrenzen. <ol><li> Führen Sie eine Reihe von Y-Labyrinth-Tests bei 25 ° C unter dunkelrotem Licht (mit LED-Lampen, um mögliche Wärmequelle zu beschränken) auf visuelle Reize zu vermeiden, so viel wie möglich. Achten Sie darauf, die Orientierungen der Y-Labyrinthe wechseln (Geruchsstoff enthalten Rohr auf der linken oder auf der rechten Seite, und Laderohr vorne oder hinten; Abbildung 1C). </li><li> Warten Sie einige Stunden für die Fliegen in der Falle Fläschchen mit dem Geruchsstoff oder Lösungsmittel ein. Graf Fliegen nach 24 Stunden, um die participati erhöhenauf bis zu mehr als 80% und bieten maximale Geruchsindexwert (Simmonet, persönliche Mitteilung). </li><li> Berechnen Sie die resultierende Geruchs Index nach folgender Formel: (Anzahl in den Geruch Rohr - Zahl in der Lösungsmittelrohr) / Gesamtzahl der geladenen Fliegen. </li></ol></li><li> Wash Y-Labyrinth Set-up wie folgt: einweichen abgebaut Set-up in RBS 35 MD Übernachtung. Gründlich mit Leitungswasser abspülen. Schließlich mit VE-Wasser spülen und trocknen lassen. </li></ol> 3. Statistische Analyse der Daten <ol><li> Führen t-Test, eine Einweg-ANOVA oder ein Zwei-Wege-ANOVA Abhängigkeit von Daten und Variablen. </li></ol>

<ol>
	<li>Mombaerts, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genes+and+ligands+for+odorant,+vomeronasal+and+taste+receptors.">Genes and ligands for odorant, vomeronasal and taste receptors.</a> Nat. Rev. Neurosci. 5, 263-278 (2004).</li><li>Silbering, A. F., Benton, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ionotropic+and+metabotropic+mechanisms+in+chemoreception:+'chance+or+design.">Ionotropic and metabotropic mechanisms in chemoreception: 'chance or design.</a> EMBO Rep. 11, 173-179 (2010).</li><li>Ziegler, A. B., Berthelot-Grosjean, M., Grosjean, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+smell+of+love+in+Drosophila.">The smell of love in Drosophila.</a> Front. Physiol. 4, (2013).</li><li>Silbering, A. F., 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=Complementary+function+and+integrated+wiring+of+the+evolutionarily+distinct+Drosophila+olfactory+subsystems.">Complementary function and integrated wiring of the evolutionarily distinct Drosophila olfactory subsystems.</a> J. Neurosci. 31, 13357-13375 (2011).</li><li>Grosjean, Y., 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=An+olfactory+receptor+for+food-derived+odours+promotes+male+courtship+in+Drosophila.">An olfactory receptor for food-derived odours promotes male courtship in Drosophila.</a> Nature. 478, 236-240 (2011).</li><li>Woodard, C., Huang, T., Sun, H., Helfand, S. L., Carlson, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+analysis+of+olfactory+behavior+in+Drosophila:+a+new+screen+yields+the+ota+mutants.">Genetic analysis of olfactory behavior in Drosophila: a new screen yields the ota mutants.</a> Genetics. 123, 315-326 (1989).</li><li>Semmelhack, J. L., Wang, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Select+Drosophila+glomeruli+mediate+innate+olfactory+attraction+and+aversion.">Select Drosophila glomeruli mediate innate olfactory attraction and aversion.</a> Nature. 459, 218-223 (2009).</li><li>Budick, S. A., Dickinson, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Free-flight+responses+of+Drosophila+melanogaster+to+attractive+odors.">Free-flight responses of Drosophila melanogaster to attractive odors.</a> The Journal of Experimental Biology. 209, 3001-3017 (2006).</li><li>Martin, F., Charro, M. J., Alcorta, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mutations+affecting+the+cAMP+transduction+pathway+modify+olfaction+in+Drosophila.">Mutations affecting the cAMP transduction pathway modify olfaction in Drosophila.</a> J. Comp. Physiol. A. 187, 359-370 (2001).</li><li>Barron, A. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anaesthetising+Drosophila+for+behavioural+studies.">Anaesthetising Drosophila for behavioural studies.</a> Journal of Insect Physiology. 46, 439-442 (2000).</li><li>Ai, M., 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=Acid+sensing+by+the+Drosophila+olfactory+system.">Acid sensing by the Drosophila olfactory system.</a> Nature. 468, 691-695 (2010).</li><li>Becher, P. G., Bengtsson, M., Hansson, B. S., Witzgall, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flying+the+fly:+long-range+flight+behavior+of+Drosophila+melanogaster+to+attractive+odors.">Flying the fly: long-range flight behavior of Drosophila melanogaster to attractive odors.</a> J. Chem. Ecol. 36, 599-607 (2010).</li></ol>

Die Autoren haben nichts zu offenbaren.

Unsere Y-Labyrinth-Protokoll basiert auf einem zuvor beschriebenen Protokoll 9 basiert. Allerdings stellen wir zwei wesentliche Unterschiede. Erstens verwenden wir engen Pipettenspitzen, um die Fliegen von der Rückkehr zu verhindern, wenn sie sich entscheiden, in der Ampulle mit dem Lösungsmittel oder Lösungsmittel plus den Geruchsstoff geben. Diese schmalen Spitzen sind auch nützlich, um die Geruchsstoffdiffusion in der Y-Labyrinth begrenzen. Zweitens verwenden wir ein kleiner Laden Fläschchen, um die F...

Chemoreception durch Geschmack oder Geruchssinn ist ein Schlüssel sensorischen Modalität für das Überleben der Tiere. Es gibt dadurch wichtige Hinweise für die Erkennung einer Gefahr oder Nahrungsquellen notwendig, als auch für soziale Interaktionen. Es hilft auch Tieren eine Sex-Partner für ihre Fortpflanzung notwendig finden. Seit mehr als 20 Jahren intensiver Forschung, einschließlich der Nobelpreis ausgezeichneten Arbeiten von Richard Axel und Linda Buck im Jahr 2004 &quot;für ihre Entdeckungen der Geruchsrezeptoren und der Organisation des olfaktorischen Systems&quot;, wurde durchgeführt, um die molekularen und zellulären Grundlagen der offenbaren Riechen 1,2. Eines der beliebtesten Tiermodelle für Wissenschaftler Geruchswahrnehmung zu zerlegen ist D. melanogaster. Dieses Insekt teilt eine ähnliche zelluläre und molekulare Geruchscodierungsstrategie mit Säugetieren. Die wissenschaftliche Gemeinschaft nutzt diverse Verhaltensparadigmen, die Rolle der Geruchsstoffe in diesem Fruchtfliege studieren. Diese Tests umfassen multimodalen Assays wie courtship Tests, bei denen verschiedene Sinnesmodalitäten, einschließlich Geruchssinn, sind wichtig, um männliche Balz 3 entlocken. Andere Tests sind auch entwickelt worden, um die Rolle von Geruchsstoffen genauer anzugehen; Dazu gehören T-Labyrinthe, Y-Labyrinthe, trap-Assays, Vier-Felder-Arenen und Wind-Tunnel 4,5,6,7,8. In diesem Artikel stellen wir eine einfache modifizierte Y-Labyrinth-Test, die robust Riech Antworten bietet mit D. melanogaster. Unser Set-up verwendet End-Tipps im Gegensatz zu einer zuvor beschriebenen Verfahren 9. So hat unsere Y-Labyrinth zwei Vorteile. Erstens, eine Rückkehr in das System vermeidet es einmal die Fliege hat ihre Wahl getroffen. Zweitens schränkt sie den Austausch von Geruchsstoffen in allen Bereichen der Y-Labyrinth. Dieser letzte Vorteil ist wichtig, da Drosophila sind sehr empfindlich auf Luftstrom, der oft verwendet wird, um Geruchs Sättigung zu vermeiden. Zur Einstellung des experimentellen Aufbaus mit einem Luftstrom würde Zeit-und kostenintensiv sein. Daher sind unsere Y-Labyrinth-Test stellt eine efreichend und schnelle Möglichkeit, Riechleistung von Drosophila zu testen.

<table><tbody><tr><td>Drosophila Polystyrene tube</td><td>VWR europe</td><td>734-2255</td><td>30 x 25 mm&amp;nbsp;&lt;br/&gt; Y-maze</td></tr><tr><td>Drosophila Borosilicate tube</td><td>Dijon verre</td><td></td><td>95 X 25 mm&lt;br/&gt; Y-maze</td></tr><tr><td>Foam stopper</td><td>Dutscher</td><td>999038</td><td>&lt;br/&gt; Y-maze</td></tr><tr><td>Y-shaped connector</td><td>Europrix</td><td>11020605</td><td>&lt;br/&gt; Y-maze</td></tr><tr><td>100-1,000 &amp;micro;l pipette tips</td><td>Corning</td><td>4868</td><td>Join the following pipette tips to the Y-shaped connector. Cut 2 pipette tips at 65 mm from the wide end, and connect the narrow end (with a &amp;sim;2 mm opening) to 2 test vials. These openings will limit the U-turns once the flies enter the tubes containing the odors. Cut 1 pipette tip at 35 mm from the wide end, and connect it to the loading vial.&lt;br/&gt; Y-maze</td></tr><tr><td>Far-Red LED Bulb</td><td>Rubin-Lacaque</td><td>0RB180238</td><td>625-630 nm</td></tr><tr><td>Acetic Acid</td><td>Sigma-Aldrich</td><td>45725</td><td></td></tr><tr><td>Phenylacetic Acid</td><td>Sigma-Aldrich</td><td>P16621</td><td></td></tr><tr><td>Yeast</td><td>Sensient Flavors Strasbourg</td><td>1018880464</td><td></td></tr><tr><td>Cornmeal</td><td>eurogerm</td><td>Farine de ma&amp;iuml;s</td><td></td></tr><tr><td>Agar</td><td>Kalys</td><td>HP-697-25</td><td></td></tr><tr><td>Methyl hydroxy 4 benzoate</td><td>VWR international</td><td>25605293</td><td></td></tr></tbody></table>

testing drosophila olfaction with a y-maze assay

Detektieren von Signalen aus der Umgebung ist für Tiere ihr Überleben zu sichern. Zu diesem Zweck verwenden sie Umweltreize wie Sehen, Mechanorezeption, Hören und chemoperception durch Geschmack, über direkten Kontakt oder durch Geruchssinn, der die Antwort auf einen flüchtigen Molekül größere Reichweite handeln darstellt. Flüchtige chemische Moleküle sind sehr wichtige Signale für die meisten Tiere in der Erkennung von Gefahr-, Nahrungsquelle oder zwischen Individuen kommunizieren. Drosophila melanogaster ist einer der am häufigsten verwendeten biologischen Modellen für Wissenschaftler, die zellulären und molekularen Basis des Geruchssinns erkunden. Um olfaktorischen Fähigkeiten dieser kleinen Insekten zu markieren, beschreiben wir eine veränderte Wahl-Protokoll basiert auf der Y-Labyrinth-Test klassisch mit Mäusen verwendet. Mit Y-Labyrinthe erhaltenen Daten geben wertvolle Informationen, um besser zu verstehen, wie Tiere mit ihren sich ständig verändernden Umwelt umzugehen. Wir führen eine Schritt-für-Schritt-Protokoll zu studieren, dieWirkung von Geruchsstoffen auf Fly Sondierungs Antwort mit diesem Y-Labyrinth-Test.

1D zeigt zwei repräsentative Antworten mit diesem Y-Labyrinth-Test. Canton-S Männer stark zu vermeiden 10% Essigsäure in destilliertem Wasser verdünnt, während sie sich nicht signifikant von 10% Phenylessigsäure vermeiden. Diese Tests werden auf 10 Männer pro Wiederholung zusammen in den Ladefläschchen gegeben basiert. Dieses Protokoll kann manchmal zu großen Standardfehler des Mittelwerts führen. Bei Bedarf ist es möglich, diesen Nachteil, indem 20 Männer pro Wiederholungen statt nur 10. Ma...

Watch this Scientific Journal Video about Testing Drosophila Olfaction with a Y-maze Assay at JoVE.com

Testing Drosophila Olfaction with a Y-maze Assay

Wir präsentieren eine Choice-Test, um den Einfluss von Geruchsstoffen auf Drosophila Verhalten mit einem Y-Labyrinth-Test offenbaren.

Testen<em&gt; Drosophila</em&gt; Riechen mit einem Y-Labyrinth-Test

Detecting signals from the environment is essential for animals to ensure their survival. To this aim, they use environmental cues such as vision, ...

testing-drosophila-olfaction-with-a-y-maze-assay

Universidad de Cartagena UDEC

Research

JoVE Journal

Neuroscience

Testing Drosophila Olfaction with a Y-maze Assay

14.8K Aufrufe.  UMR-6265 CNRS.  Wir präsentieren eine Choice-Test, um den Einfluss von Geruchsstoffen auf Drosophila Verhalten mit einem Y-Labyrinth-Test offenbaren. 

Serie: Testing Drosophila Olfaction with a Y-maze Assay

High-resolution Measurement of Odor-Driven Behavior in Drosophila Larvae

Olfactory responses in Drosophila larvae have been traditionally studied in Petri dishes comprising a single peripheral odor source. In this behavioral paradigm, the experimenter usually assumes that the rapid diffusion of odorant molecules from the source leads to the creation of a stable gradient in the dish. To establish a quantitative correlation between sensory inputs and behavioral responses, it is necessary to achieve a more thorough characterization of the odorant stimulus conditions. In this video article, we describe a new method allowing the construction of odorant gradients with stable and controllable geometries. We briefly illustrate how these gradients can be used to screen for olfactory defects (full and partial anosmia) and to study more subtle features of chemotaxis behavior.

In this video article, we describe a new method allowing the construction of odorant gradients with stable and controllable geometries. We briefly illustrate how these gradients can be used to screen for olfactory defects (full and partial anosmia) and to study more subtle features of chemotaxis behavior.

Hochauflösende Messung von Odor-Driven Behavior in Drosophila-Larven

Olfactory responses in Drosophila larvae have been traditionally studied in Petri dishes comprising a single peripheral odor source. In this behavioral ...

Forschung

Biologie

Drosophila Adult Olfactory Shock Learning

Drosophila have been used in classical conditioning experiments for over 40 years, thus greatly facilitating our understanding of memory, including the elucidation of the molecular mechanisms involved in cognitive diseases1-7. Learning and memory can be assayed in larvae to study the effect of neurodevelopmental genes8-10 and in flies to measure the contribution of adult plasticity genes1-7. Furthermore, the short lifespan of Drosophila facilitates the analysis of genes mediating age-related memory impairment5,11-13. The availability of many inducible promoters that subdivide the Drosophila nervous system makes it possible to determine when and where a gene of interest is required for normal memory as well as relay of different aspects of the reinforcement signal3,4,14,16.
Studying memory in adult Drosophila allows for a detailed analysis of the behavior and circuitry involved and a measurement of long-term memory15-17. The length of the adult stage accommodates longer-term genetic, behavioral, dietary and pharmacological manipulations of memory, in addition to determining the effect of aging and neurodegenerative disease on memory3-6,11-13,15-21.
Classical conditioning is induced by the simultaneous presentation of a neutral odor cue (conditioned stimulus, CS+) and a reinforcement stimulus, e.g., an electric shock or sucrose, (unconditioned stimulus, US), that become associated with one another by the animal1,16. A second conditioned stimulus (CS-) is subsequently presented without the US. During the testing phase, Drosophila are simultaneously presented with CS+ and CS- odors. After the Drosophila are provided time to choose between the odors, the distribution of the animals is recorded. This procedure allows associative aversive or appetitive conditioning to be reliably measured without a bias introduced by the innate preference for either of the conditioned stimuli. Various control experiments are also performed to test whether all genotypes respond normally to odor and reinforcement alone.

Drosophila Adult Olfactory Shock Learning

The method to measure adult Drosophila associative memory is described. The assay is based on the ability of the fly to associate an odor presented with a negative reinforcer (electric shock) and then recall this information at a later time, allowing memory to be measured.

Drosophila Erwachsene olfaktorischen Schock Learning

Drosophila have been used in classical conditioning experiments for over 40 years, thus greatly facilitating our understanding of memory, including the ...

Neurowissenschaften

Vertical T-maze Choice Assay for Arthropod Response to Odorants

Given the economic importance of insects and arachnids as pests of agricultural crops, urban environments or as vectors of plant and human diseases, various technologies are being developed as control tools. A subset of these tools focuses on modifying the behavior of arthropods by attraction or repulsion. Therefore, arthropods are often the focus of behavioral investigations. Various tools have been developed to measure arthropod behavior, including wind tunnels, flight mills, servospheres, and various types of olfactometers. The purpose of these tools is to measure insect or arachnid response to visual or more often olfactory cues. The vertical T-maze oflactometer described here measures choices performed by insects in response to attractants or repellents. It is a high throughput assay device that takes advantage of the positive phototaxis (attraction to light) and negative geotaxis (tendency to walk or fly upward) exhibited by many arthropods. The olfactometer consists of a 30 cm glass tube that is divided in half with a Teflon strip forming a T-maze. Each half serves as an arm of the olfactometer enabling the test subjects to make a choice between two potential odor fields in assays involving attractants. In assays involving repellents, lack of normal response to known attractants can also be measured as a third variable.

A vertical, T-maze olfactometer is described for assaying the behavioral response of arthropods. The olfactometer allows the experimenter to measure choices performed by test subjects when subjected to two potential odor fields. Both attraction to and repulsion from odorants can be measured with this device.

Vertical T-Labyrinth Wahl Assay für Gliederfüßer Response to Geruchsstoffe

Given the economic importance of insects and arachnids as pests of agricultural crops, urban environments or as vectors of plant and human diseases, ...

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.

Ein Single-fly-Assay für Sammelverhalten 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 ...

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.

Einfache Assay zur Quantifizierung von Sozialvermeidung 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 ...

High-resolution Quantification of Odor-guided Behavior in Drosophila melanogaster Using the Flywalk Paradigm

In their natural environment, insects such as the vinegar fly Drosophila melanogaster are bombarded with a huge amount of chemically distinct odorants. To complicate matters even further, the odors detected by the insect nervous system usually are not single compounds but mixtures whose composition and concentration ratios vary. This leads to an almost infinite amount of different olfactory stimuli which have to be evaluated by the nervous system.
To understand which aspects of an odor stimulus determine its evaluation by the fly, it is therefore desirable to efficiently examine odor-guided behavior towards many odorants and odor mixtures. To directly correlate behavior to neuronal activity, behavior should be quantified in a comparable time frame and under identical stimulus conditions as in neurophysiological experiments. However, many currently used olfactory bioassays in Drosophila neuroethology are rather specialized either towards efficiency or towards resolution.
Flywalk, an automated odor delivery and tracking system, bridges the gap between efficiency and resolution. It allows the determination of exactly when an odor packet stimulated a freely walking fly, and to determine the animal&#180;s dynamic behavioral reaction.

High-resolution Quantification of Odor-guided Behavior in Drosophila melanogaster Using the Flywalk Paradigm

The automated tracking system Flywalk is used for high-resolution quantification of odor-guided behavior in Drosophila melanogaster.

Hochaufgelöste Quantifizierung der Geruchsgelenkten Verhalten in<em&gt; Drosophila melanogaster</em&gt; Verwenden der<em&gt; Flywalk</em&gt; Paradigm

In their natural environment, insects such as the vinegar fly Drosophila melanogaster are bombarded with a huge amount of chemically distinct odorants. To ...

Olfactory Behaviors Assayed by Computer Tracking Of Drosophila in a Four-quadrant Olfactometer

A key challenge in neurobiology is to understand how neural circuits function to guide appropriate animal behaviors. Drosophila melanogaster is an excellent model system for such investigations due to its complex behaviors, powerful genetic techniques, and compact nervous system. Laboratory behavioral assays have long been used with Drosophila to simulate properties of the natural environment and study the neural mechanisms underlying the corresponding behaviors (e.g. phototaxis, chemotaxis, sensory learning and memory)1-3. With the recent availability of large collections of transgenic Drosophila lines that label specific neural subsets, behavioral assays have taken on a prominent role to link neurons with behaviors4-11. Versatile and reproducible paradigms, together with the underlying computational routines for data analysis, are indispensable for rapid tests of candidate fly lines with various genotypes. Particularly useful are setups that are flexible in the number of animals tested, duration of experiments and nature of presented stimuli. The assay of choice should also generate reproducible data that is easy to acquire and analyze. Here, we present a detailed description of a system and protocol for assaying behavioral responses of Drosophila flies in a large four-field arena. The setup is used here to assay responses of flies to a single olfactory stimulus; however, the same setup may be modified to test multiple olfactory, visual or optogenetic stimuli, or a combination of these. The olfactometer setup records the activity of fly populations responding to odors, and computational analytical methods are applied to quantify fly behaviors. The collected data are analyzed to get a quick read-out of an experimental run, which is essential for efficient data collection and the optimization of experimental conditions.

Olfactory Behaviors Assayed by Computer Tracking Of Drosophila in a Four-quadrant Olfactometer

We describe here a behavioral setup and data analysis method for assaying olfactory responses of up to 100 vinegar flies (Drosophila melanogaster). This system may be used with single or multiple olfactory stimuli, and adaptable for optogenetic activation or silencing of neuronal subsets.

Olfaktorischen Behaviors Assayed von Computer-Tracking<em&gt; Drosophila</em&gt; In einem Vier-Quadranten-Olfaktometer

A key challenge in neurobiology is to understand how neural circuits function to guide appropriate animal behaviors. Drosophila melanogaster is an ...

Taste Preference Assay for Adult Drosophila

Olfactory and gustatory perception of the environment is vital for animal survival. The most obvious application of these chemosenses is to be able to distinguish good food sources from potentially dangerous food sources. Gustation requires physical contact with a chemical compound which is able to signal through taste receptors that are expressed on the surface of neurons. In insects, these gustatory neurons can be located across the animal's body allowing taste to play an important role in many different behaviors. Insects typically prefer compounds containing sugars, while compounds that are considered bitter tasting are avoided. Given the basic biological importance of taste, there is intense interest in understanding the molecular mechanisms underlying this sensory modality. We describe an adult Drosophila taste assay which reflects the preference of the animals for a given tastant compound. This assay may be applied to animals of any genetic background to examine the taste preference for a desired soluble compound.

Taste Preference Assay for Adult Drosophila

Taste is an important sensory process which facilitates attraction to beneficial substances and avoidance of toxic substances. This protocol describes a simple ingestion assay for determining Drosophila gustatory preference for a given chemical compound.

Geschmackspräferenz Assay für Erwachsene<em&gt; Drosophila</em

Olfactory and gustatory perception of the environment is vital for animal survival.  The most obvious application of these chemosenses is to be able to ...

Drosophila Modelle für genetische und verhaltensbezogene Merkmale der Autismus-Spektrum-Störung

Paradigms for Behavioral Assessment in Drosophila Model of Autism Spectrum Disorder

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

Autor im Rampenlicht: Erforschung der Symptome der Autismus-Spektrum-Störung bei Fruchtfliegen – genetische Modelle und Verhaltenstests

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

Paradigmen für die Verhaltensbewertung im Drosophila-Modell der Autismus-Spektrum-Störung

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

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