Agradecemos 4 evaluadores anónimos por su trabajo para mejorar el manuscrito. Agradecemos al Centro Nacional de Investigación Científica por su apoyo financiero a MBG y YG, y la Université de Bourgogne y el Ministerio francés de Investigación para MMS. La investigación en laboratorio YG es financiado por el Consejo Europeo de Investigación (ERC Starting Grant, GliSFCo-311403), la Agence Nationale de la Recherche (ANR-JCJC, GGCB-2010), el Conseil Régional de Bourgogne (Faber) y el CNRS.

1. Antes de empezar <ol><li> Utilizar una referencia de valores isogenized teniendo fenotipos conductuales estables y robustas. No existe una regla general para la elección de esta población, ya que todos los controles posibles pueden llevar heterogéneos alelos de fondo. </li><li> Utilice esta cepa de control para retrocruzar cada dos valores necesarios para los pasos posteriores. Este paso retrocruzamiento se representa típicamente por lo menos 5 cruces sucesivos de una sola hembra virgen (para permitir que sea posible entrecruzamiento entre cromosomas homólogos) a 2-3 machos de referencia isogénicas 5. Este paso es importante para homogeneizar el fondo genético entre las diferentes poblaciones de la mosca. </li><li> Mantener las poblaciones de Drosophila en una harina de maíz estándar (9%), levadura (10%), y el medio de agar (1,5%) complementado con antibióticos (0,4% de metil para-hidroxi-benzoato) en un ciclo de luz / oscuridad de 12 horas a 25 ° C. </li><li> Lograr experimentos quimiosensoriales en una habitación con temperatura controlada (25 º C) con luz roja lejana(Para eliminar la contribución de las señales visuales, y centrarse en las señales quimiosensoriales). Regularmente renovar el aire de la habitación a ventilar el área entre cada experimento. </li></ol> 2. Respuesta olfativa mediante un ensayo del laberinto <ol><li> Morir de hambre a las moscas de 16-18 horas a 25 ° C en tubos de vidrio que contienen una toalla de papel húmeda antes de la prueba. </li><li> Únase a un conector en forma de Y a dos viales de vidrio y un vial de plástico pequeña (carga vial). Utilice 1 ml puntas de pipeta que pasan a través de los tapones de espuma para unir el conector a los tres viales, y para obtener un laberinto Y cerrado herméticamente. Cortar los extremos estrechos de dos puntas de pipeta (~ 2 mm de diámetro, para evitar cualquier retorno de la marcha una vez que ha tomado su decisión) para formar dos viales &quot;atrapar&quot;, y un gran final de una punta de la pipeta para formar la &quot;carga&quot; tubo (Figura 1A). </li><li> Justo antes de la conexión de los viales &quot;atrapar&quot; (fig. 1B), coloque una ~ 6 mm de diámetro en el papel de filtrocada vial. Añadir 40 l de solución de odorante en una de papel de filtro, y 40 l de la correspondiente disolvente en el segundo papel de filtro. </li><li> Introducir diez 4-9 moscas de un día en el vial de &quot;carga&quot;. No utilice CO 2 anestesia durante esta transferencia, ya que tiene un fuerte efecto sobre el comportamiento 10. Más bien utilizar breve enfriamiento en hielo. La manipulación adecuada de las moscas anestesiadas es importante limitar el estrés en los temas tanto como sea posible. <ol><li> Lleve a cabo una serie de pruebas del laberinto a 25 ° C bajo luz roja lejana (el uso de bombillas LED para limitar la posible fuente de calor) para evitar los estímulos visuales tanto como sea posible. Tenga cuidado de alternar las orientaciones de los Y-laberintos (tubo odorante que contiene a la izquierda, oa la derecha, y el tubo de carga en el frente o en la espalda; Figura 1C). </li><li> Permita que transcurran varias horas para que las moscas entren en el vial que contiene la trampa odorizante o el disolvente. Cuente moscas después de 24 horas para aumentar la participatien un máximo de más de 80% y proporcionar el valor de índice olfativa máxima (Simonnet, comunicación personal). </li><li> Calcular el índice olfativa resultante usando la siguiente fórmula: (número en el tubo de olor - número en el tubo de disolvente) / número total de moscas cargados. </li></ol></li><li> Lave laberinto Y puesta a punto de la siguiente manera: remoje la desmantelado configuración en RBS 35 MD durante la noche. Enjuagar bien con agua del grifo. Por último, enjuagar con agua desionizada y dejar que se seque. </li></ol> 3. Análisis Estadístico de los Datos <ol><li> Lleve a cabo una prueba t, un ANOVA de una vía o un ANOVA de dos vías en función de los datos y variables. </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>

Los autores no tienen nada que revelar.

Nuestro protocolo laberinto Y se basa en un protocolo previamente descrito 9. Sin embargo, se introducen dos grandes diferencias. En primer lugar, utilizamos puntas de pipeta estrechos para evitar que las moscas de regresar una vez que deciden entrar en el vial que contiene el disolvente o el disolvente más el odorante. Estos consejos estrechos también son útiles para limitar la difusión de aroma en el laberinto. En segundo lugar, se utiliza un vial de carga menor a la fuerza a las moscas para entrar en e...

Quimiorrecepción a través del gusto o el olfato es una modalidad sensorial clave para la supervivencia de los animales. Se da señales vitales necesarias para la detección de un peligro o de las fuentes de alimentos, así como para las interacciones sociales. También ayuda a los animales a encontrar un socio necesario el sexo para su reproducción. Durante más de 20 años, la investigación intensiva, incluyendo el premio Nobel de obra de Richard Axel y Linda Buck en 2004 &quot;por sus descubrimientos de los receptores odorantes y la organización del sistema olfativo&quot;, se ha llevado a cabo para revelar las bases moleculares y celulares de la olfacción 1,2. Uno de los modelos animales preferidos por los científicos para diseccionar la percepción olfativa es D. melanogaster. Este insecto comparte una estrategia de codificación de olor celular y molecular similar con mamíferos. La comunidad científica utiliza diversos paradigmas de comportamiento para estudiar el papel de los odorantes en esta mosca de la fruta. Estas pruebas incluyen ensayos multimodales como courtshipruebas p donde diversas modalidades sensoriales, incluyendo el olfato, son importantes para obtener el cortejo masculino 3. Otros ensayos también se han desarrollado para abordar el papel de los odorantes más específicamente; estos incluyen T-laberintos, Y-laberintos, ensayos de trampa, arenas de cuatro de campo y 4.5.6.7.8 viento túneles. En este artículo se presenta un ensayo del laberinto sencillo modificado, que proporciona respuestas olfativas robustas usando D. melanogaster. Nuestra puesta a punto utiliza-tips finales en contra de un método descrito anteriormente 9. Por lo tanto, nuestra laberinto Y tiene dos ventajas. En primer lugar, evita las subidas en el sistema una vez que la marcha ha hecho su elección. En segundo lugar, limita el intercambio de olores en todas las áreas de el laberinto. Esta última ventaja es importante, ya que de Drosophila son muy sensibles al flujo de aire que se utiliza a menudo para evitar la saturación odorante. Para ajustar el montaje experimental con un flujo de aire sería consume tiempo y costo. Por lo tanto, nuestro ensayo del laberinto representa un efciente y rápida para poner a prueba el rendimiento olfativo de Drosophila.

<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

La detección de las señales del medio ambiente es esencial para los animales para asegurar su supervivencia. Para ello, utilizan señales ambientales tales como la visión, mecanorrecepción, el oído y chemoperception a través del gusto, por contacto directo oa través del olfato, el cual representa la respuesta a una molécula volátil que actúa en un mayor alcance. Moléculas químicas volátiles son señales muy importantes para la mayoría de los animales en la detección de peligro, una fuente de alimento, o para la comunicación entre individuos. Drosophila melanogaster es uno de los modelos biológicos más comunes para los científicos a explorar la base celular y molecular del olfato. Con el fin de resaltar las habilidades olfativas de este pequeño insecto, se describe un protocolo elección modificado basada en la prueba del laberinto en clásicamente utilizado con ratones. Los datos obtenidos con Y-laberintos dan información valiosa para comprender mejor cómo tratan los animales con su entorno permanentemente cambiante. Introducimos un protocolo paso a paso para estudiar laimpacto de los olores en mosca respuesta exploratorio utilizando este ensayo del laberinto.

Figura 1D muestra dos respuestas representativas utilizando este ensayo del laberinto. Cantón-S machos evitan fuertemente 10% de ácido acético diluido en agua destilada, mientras que no evitan significativamente ácido fenilacético 10%. Estos ensayos se basan en 10 varones por réplica colocados juntos en los viales de carga. Este protocolo a veces puede conducir a grandes errores estándar de la media. Si es necesario, es posible reducir este inconveniente mediante el uso de 20 hombres por repetici...

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

Se presenta una prueba de elección para revelar la influencia de sustancias odoríferas en el comportamiento de Drosophila utilizando un ensayo del laberinto.

Pruebas<em&gt; Drosophila</em&gt; El olfato con un ensayo del laberinto

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 vistas.  UMR-6265 CNRS.  Se presenta una prueba de elección para revelar la influencia de sustancias odoríferas en el comportamiento de Drosophila utilizando un ensayo del laberinto. 

Video: 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.

De alta resolución de medición del comportamiento de olor-Driven en larvas de Drosophila

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

Investigación

Biología

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 Adulto olfativa Choque Aprendizaje

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

Neurociencias

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-laberinto Choice Ensayo para la respuesta a los olores artrópodos

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.

Un ensayo de una sola mosca por el comportamiento de forrajeo en<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.

Ensayo sencillo para cuantificación de Prevención 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 ...

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.

De alta resolución de cuantificación del comportamiento guiado Olor en<em&gt; Drosophila melanogaster</em&gt; Uso de la<em&gt; Flywalk</em&gt; Paradigma

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.

Comportamientos olfativas ensayó mediante seguimiento informático de<em&gt; Drosophila</em&gt; En un olfatómetro de cuatro cuadrantes

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.

Ensayo Preferencia gusto por adulto<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 Modelos de rasgos genéticos y conductuales del trastorno del espectro autista

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 destacado: Explorando los síntomas del trastorno del espectro autista en las moscas de la fruta: modelos genéticos y pruebas de comportamiento

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.

Paradigmas para la evaluación conductual en el modelo de Drosophila del trastorno del espectro autista

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

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