Name: a method to test the effect of environmental cues on mating behavior in drosophila melanogaster
Uploaded: 2017-07-17T13:00:00.000+00:00
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Description: Watch this Scientific Journal Video about A Method to Test the Effect of Environmental Cues on Mating Behavior in Drosophila melanogaster at JoVE.com

Damos las gracias al Bloomington Drosophila Stock Center para las acciones de la mosca; C. Gahr, JT Alkema y S. van Hasselt por su temprano intento de desarrollar el ensayo de aire presurizado; Jasper Bosman por el consejo sobre cultivar levadura; Y Rezza Azanchi y Joel Levine por haber desarrollado originalmente el monitoreo de lapso de tiempo del comportamiento de apareamiento de Drosophila . JA Gorter recibió el apoyo de una Beca del Programa de Posgrado BCN / NWO de la Escuela de Investigación Neurocientífica. Este trabajo fue apoyado en parte por la organización holandesa para la investigación científica (NWO) (referencia: 821.02.020) a JC Billeter.

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Este protocolo describe un ensayo para probar el comportamiento de apareamiento durante 24 horas mientras se controlan continuamente las señales ambientales que se supone que una pareja conjugada usa para determinar la frecuencia de apareamiento. Es posible aumentar la frecuencia de apareamiento en respuesta al aire de levadura suministrado a través de un sistema de aire presurizado cuando el medio contiene levadura también ( Figura 2B ). Además, se puede observar una respuesta similar en la frecuen...

Los comportamientos reproductivos suelen tener altos costos de energía, especialmente para las hembras, que producen gametos más grandes que los machos y deben escoger cuidadosamente las condiciones para criar a sus descendientes en desarrollo. Debido al costo energético, no es de extrañar que la reproducción esté relacionada con las condiciones nutricionales. Esto es cierto en la mayoría de los animales, incluso en los mamíferos, cuya pubertad puede ser retardada por la desnutrición y cuyo impulso sexual puede verse afectado negativamente por la restricción de los alimentos 1 . La reproducción del organismo genético modelo Drosophila melanogaster también se ve afectada por las condiciones nutricionales. Los varones corte en el nivel más alto en la presencia de los volátiles del alimento 2 , y las hembras son sexualmente más receptivas en presencia de la levadura, un nutriente importante para la producción del huevo y la supervivencia de la descendencia 3 , 4 , 5 . EstaLa respuesta reproductiva evolutiva conservada a los alimentos ofrece la oportunidad de estudiar mecanismos que conectan la disponibilidad de alimentos ambientales a la reproducción sexual en un organismo geneticamente manejable y eficiente en el tiempo. De hecho, el trabajo en D. melanogaster ha implicado la vía de la insulina como un importante regulador de la conexión entre la comida y el comportamiento de acoplamiento [ 6] . También ha demostrado que el acto de apareamiento a sí mismo cambia la preferencia alimentaria de las hembras, así como las neuronas quimiosensoriales asociadas 7 , 8 , 9 . Está claro que las señales alimentarias afectan los comportamientos reproductivos en D. melanogaster . Estos efectos parecen afectar principalmente a las hembras, específicamente a las que ya se han apareado 5 . Sin embargo, para probar estos efectos agudos de las condiciones ambientales, el ensayo clásicamente utilizado para el comportamiento de apareamiento femenino podríaNo ser muy adecuado debido a las largas interrupciones entre los episodios de apareamiento. En el clásico ensayo de remate, una hembra virgen se aparea primero con un macho, y se aísla inmediatamente y se presenta con un nuevo macho 24 a 48 h después. Este ensayo clásico se ha utilizado con gran éxito para identificar componentes del eyaculado masculino que modifican el comportamiento femenino y la respuesta femenina 12 , 13 , 14 , 15 , 16 , 17 , 18 . El ensayo de apareamiento continuo demostrado aquí es, por lo tanto, una adición a los ensayos de apareamiento clásicos que pueden usarse para estudiar el efecto agudo de las condiciones ambientales en los comportamientos reproductivos. Utilizando el ensayo continuo para el comportamiento de apareamiento que se explica aquí, hemos demostrado previamente que un par de moscas expuestas a la levadura remate sCada vez durante un período de observación de 24 h 5 , 19 , 20 , 21 , mientras que las moscas no expuestas a los alimentos sólo remate una vez 5 . Este hallazgo puede ser desconcertante a la luz de una gran parte de la literatura de D. melanogaster que indica que las hembras no remate durante varios días después de un apareamiento inicial (revisado en las referencias 10 , 11 ). Sin embargo, esta discrepancia puede ser fácilmente explicada por condiciones de ensayo, donde una hembra se aísla de uno a varios días antes de que se proporcione una nueva oportunidad de apareamiento. Si la pareja no se aparea en este período de observación de una hora, la hembra se caracteriza por no ser receptiva. Por otra parte, la alta frecuencia de apareamiento no debe ser sorprendente, dado que los datos de las moscas capturadas en el medio silvestre muestran que las hembras contienen espermatozoides de 4 a 6 machos en sus órganos de almacenamiento; Por lo tanto enIndicando que las hembras naturalmente remate varias veces 22 , 23 . Aquí, demostrar el uso de este ensayo de apareamiento continuo para desentrañar cómo las moscas recopilar y combinar la información sobre las condiciones ambientales para modular la frecuencia de apareamiento. Este ensayo permite probar un número relativamente grande de parejas de apareamiento para estudios genéticos y probar la influencia de señales ambientales volátiles y no volátiles. El ensayo típicamente dura 24 h, pero se puede extender hasta 48 h, permitiendo la prueba de ciclos ambientales como el ciclo luz-oscuridad (LD). Demostramos este ensayo mediante la prueba de la influencia de las señales volátiles de un cultivo de levadura dentro de un sistema de aire presurizado en combinación con la disponibilidad de nutrientes no volátiles de levadura en el sustrato de alimentos. El sistema de aire presurizado bombea continuamente señales volátiles en una arena de acoplamiento que contienenUn sustrato alimenticio y una pareja de prueba (cuyo comportamiento de apareamiento es controlado). Para determinar adicionalmente las especificidades a través de las cuales la levadura influye en el apareamiento, probamos un compuesto volátil principal de levadura, a saber ácido acético 24 , en combinación con un contenido de aminoácidos que corresponde al de la levadura en el sustrato alimenticio, en forma de peptona Ácidos derivados de la digestión enzimática de proteínas animales). Juntos estos experimentos demuestran cómo el efecto de señales ambientales en el comportamiento de apareamiento de D. melanogaster puede ser probado con este ensayo.

<table><tbody><tr><td>&lt;strong&gt;Cabinet&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Stainless steel kitchen cabinet</td><td>Horecaworld</td><td>7412.0105</td><td></td></tr><tr><td>White LEDs</td><td>Lucky Light</td><td>ll-583wc2c-001</td><td>Cold white, 20 mAmp and 2 V</td></tr><tr><td>Red LEDs</td><td>Lucky Ligt</td><td>ll-583vc2c-v1-4da</td><td>Wavelength between 625 nm, 20 mAmp and 6 V</td></tr><tr><td>Resistor</td><td>Royal Ohm</td><td>CFR0W4J0561A50</td><td>560 ohm, 0.25 W, 250 V and 5 % tolerance</td></tr><tr><td>Smartphone light meter app</td><td>Patrick Giudicelli</td><td></td><td>Light/Lux Meter FREE, version 1.1.1</td></tr><tr><td>Power timer</td><td>Alecto</td><td>TS-121</td><td></td></tr><tr><td>Metal brackets</td><td></td><td></td><td>Sharp angle 5 by 5 mm,&amp;nbsp; 2 x 5450 and 1 x 1100 mm long</td></tr><tr><td>Frosted glass plate</td><td></td><td></td><td>1190 x 545 x 5 mm</td></tr><tr><td>Filter paper sheets</td><td>LEE 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yeast</td><td>Red Star</td><td></td><td></td></tr><tr><td>Tegosept</td><td>Flystuff</td><td>20-258</td><td>100%</td></tr><tr><td>Peptone (bacto)</td><td>BD</td><td>211677</td><td></td></tr><tr><td>Acetic Acid</td><td>Merck</td><td>1000631000</td><td>Glacial, 100%</td></tr><tr><td>Small petridish</td><td>Greiner bio-one</td><td>627102</td><td>35 x 10 mm with vents</td></tr><tr><td>Paraffin film</td><td>Bemis NA</td><td></td><td>Parafilm</td></tr><tr><td>&lt;strong&gt;Name&lt;/strong&gt;</td><td>&lt;strong&gt;Company&lt;/strong&gt;</td><td>&lt;strong&gt;Catalog Number&lt;/strong&gt;</td><td>&lt;strong&gt;Comments&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;Yeast and pressurised air set-up&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Big petridish</td><td>Gosselin</td><td>BP140-01</td><td>140 x 20.6 mm</td></tr><tr><td>Ultrapure water</td><td>Millipore corporation</td><td></td><td>MiliQ</td></tr><tr><td>Yeast extract</td><td>BD</td><td>212750</td><td></td></tr><tr><td>Agar (pure)</td><td>BD</td><td>214530</td><td>bacto</td></tr><tr><td>Glucose (0(+)-glucose monohydrate)&amp;nbsp;</td><td>Merck</td><td>18270000004</td><td></td></tr><tr><td>Open caps</td><td>Schott</td><td>29 240 28&amp;nbsp;</td><td>GL45</td></tr><tr><td>Silicone septum</td><td>VWR</td><td>548-0662</td><td></td></tr><tr><td>Barbed bulkhead fittings</td><td>Nalgene</td><td>6149-0002</td><td></td></tr><tr><td>Large PVC tubing</td><td></td><td></td><td>diameter: outer 1.2 cm and inner 0.9 cm</td></tr><tr><td>Small PVC tubing</td><td></td><td></td><td>diameters: outer 0.8 cm and inner 0.5 cm</td></tr><tr><td>15 ml tube</td><td>Falcon</td><td></td><td></td></tr><tr><td>Aquarium pump</td><td>Sera precision</td><td></td><td>Sera air 110 plus, AC 220-240 V, 50/60 Hz, 3 W and pressure &gt;100 mbar</td></tr><tr><td>Activated charcoal</td><td>Superfish</td><td>A8040400</td><td>Norit activated carbon</td></tr><tr><td>Disposible filter unit</td><td>Whatman</td><td>10462100</td><td></td></tr><tr><td>Serological pipettes</td><td>VWR</td><td>612-1600</td><td></td></tr><tr><td>Syringe</td><td>BD Plastipak</td><td>300013</td><td></td></tr><tr><td>Hot glue</td><td>Pattex</td><td></td><td></td></tr><tr><td>Syringe filter</td><td>Whatman</td><td></td><td>FP 30/pore size 0.45 mm CA-S</td></tr><tr><td>&lt;strong&gt;Name&lt;/strong&gt;</td><td>&lt;strong&gt;Company&lt;/strong&gt;</td><td>&lt;strong&gt;Catalog Number&lt;/strong&gt;</td><td>&lt;strong&gt;Comments&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;Analysis&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Statistics software</td><td>R</td><td></td><td>lme4 package</td></tr></tbody></table>

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

El impulso sexual de un individuo está influenciado por el genotipo, la experiencia y las condiciones ambientales. Cómo estos factores interactúan para modular comportamientos sexuales sigue siendo mal entendido. En Drosophila melanogaster , las señales ambientales, como la disponibilidad de alimentos, afectan la actividad de apareamiento, ofreciendo un sistema manejable para investigar los mecanismos que modulan el comportamiento sexual. En D. melanogaster , las señales ambientales a menudo se perciben a través de los sistemas gustativo y olfativo quimiosensoriales. Aquí, presentamos un método para probar el efecto de las señales químicas ambientales en el comportamiento de apareamiento. El ensayo consiste en una pequeña arena de apareamiento que contiene medio de alimento y una pareja de apareamiento. La frecuencia de apareamiento para cada pareja se controla continuamente durante 24 h. Aquí presentamos la aplicabilidad de este ensayo para probar compuestos ambientales de una fuente externa a través de un sistema de aire presurizado, así como la manipulación de los componentes ambientales directamente en el campo de apareamiento. El uDe un sistema de aire a presión es especialmente útil para probar el efecto de compuestos muy volátiles, mientras que la manipulación de componentes directamente en el campo de acoplamiento puede ser de valor para determinar la presencia de un compuesto. Este ensayo puede adaptarse para responder a preguntas sobre la influencia de las señales genéticas y ambientales en el comportamiento de apareamiento y la fecundidad, así como otros comportamientos reproductivos masculinos y femeninos.

Usando este ensayo continuo, el comportamiento de apareamiento, y la frecuencia de apareamiento en específico, se puede determinar bajo condiciones ambientales experimentales. Para controlar las condiciones ambientales, transformamos un gabinete de cocina de acero inoxidable en un área de prueba, con su propia fuente de luz y difusión, lo que garantiza una gran cantidad de luz y una mínima cantidad de reflejos desde la parte superior de las arenas de apareamiento ( Figura 1A<...

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

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

Se demuestra un ensayo para analizar las señales ambientales y genéticas que influyen en el comportamiento de apareamiento en la mosca de la fruta Drosophila melanogaster .

Un método para probar el efecto de las señales ambientales sobre el comportamiento de apareamiento en<em&gt; Drosophila melanogaster</em

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

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

Universidad de Cartagena UDEC

Research

JoVE Journal

Behavior

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

9.0K vistas.  University of Groningen.  Se demuestra un ensayo para analizar las señales ambientales y genéticas que influyen en el comportamiento de apareamiento en la mosca de la fruta Drosophila melanogaster . 

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

Methods to Assay Drosophila Behavior

Drosophila melanogaster, the fruit fly, has been used to study molecular mechanisms of a wide range of human diseases such as cancer, cardiovascular disease and various neurological diseases1. We have optimized simple and robust behavioral assays for determining larval locomotion, adult climbing ability (RING assay), and courtship behaviors of Drosophila. These behavioral assays are widely applicable for studying the role of genetic and environmental factors on fly behavior. Larval crawling ability can be reliably used for determining early stage changes in the crawling abilities of Drosophila larvae and also for examining effect of drugs or human disease genes (in transgenic flies) on their locomotion. The larval crawling assay becomes more applicable if expression or abolition of a gene causes lethality in pupal or adult stages, as these flies do not survive to adulthood where they otherwise could be assessed. This basic assay can also be used in conjunction with bright light or stress to examine additional behavioral responses in Drosophila larvae. Courtship behavior has been widely used to investigate genetic basis of sexual behavior, and can also be used to examine activity and coordination, as well as learning and memory. Drosophila courtship behavior involves the exchange of various sensory stimuli including visual, auditory, and chemosensory signals between males and females that lead to a complex series of well characterized motor behaviors culminating in successful copulation. Traditional adult climbing assays (negative geotaxis) are tedious, labor intensive, and time consuming, with significant variation between different trials2-4. The rapid iterative negative geotaxis (RING) assay5 has many advantages over more widely employed protocols, providing a reproducible, sensitive, and high throughput approach to quantify adult locomotor and negative geotaxis behaviors. In the RING assay, several genotypes or drug treatments can be tested simultaneously using large number of animals, with the high-throughput approach making it more amenable for screening experiments.

Methods to Assay Drosophila Behavior

Drosophila melanogaster is a genetically and behaviorally tractable model system that has been used to understand the molecular and cellular basis of many important biological processes for over a century 1. Drosophila has been well exploited to gain insights into the genetic basis of fly behavior.

Métodos de ensayo a Drosophila Comportamiento

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

Investigación

Neurociencias

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 ...

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.

Teñido Insectos de la Conducta ensayos: el comportamiento de apareamiento de anestesiada<em&gt; Drosophila</em

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

Conditions Affecting Social Space in Drosophila melanogaster

The social space assay described here can be used to quantify social interactions of Drosophila melanogaster &#8212; or other small insects &#8212; in a straightforward manner. As we previously demonstrated 1, in a two-dimensional chamber, we first force the flies to form a tight group, subsequently allowing them to take their preferred distance from each other. After the flies have settled, we measure the distance to the closest neighbor (or social space), processing a static picture with free online software (ImageJ). The analysis of the distance to the closest neighbor allows researchers to determine the effects of genetic and environmental factors on social interaction, while controlling for potential confounding factors. Diverse factors such as climbing ability, time of day, sex, and number of flies, can modify social spacing of flies. We thus propose a series of experimental controls to mitigate these confounding effects. This assay can be used for at least two purposes. First, researchers can determine how their favorite environmental shift (such as isolation, temperature, stress or toxins) will impact social spacing 1,2. Second, researchers can dissect the genetic and neural underpinnings of this basic form of social behavior 1,3. Specifically, we used it as a diagnostic tool to study the role of orthologous genes thought to be involved in social behavior in other organisms, such as candidate genes for autism in humans 4.

Conditions Affecting Social Space in Drosophila melanogaster

The effect of genes and environment on social space of Drosophila melanogaster can be quantified through a powerful but straightforward analytical paradigm. We show here different factors that can affect this social space, and thus need to be taken into consideration when designing experiments in this paradigm.

Condiciones que afectan Espacio Social en<em&gt; Drosophila melanogaster</em

The social space assay described here can be used to quantify social interactions of Drosophila melanogaster &#8212; or other small insects &#8212; in a ...

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

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

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

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

Un nuevo enfoque que elimina el manejo para el estudio de la agresión y el efecto &quot;Loser&quot; en<em&gt; Drosophila melanogaster</em

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

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

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

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

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

Un ensayo de conducta para Mechanosensation de clones en MARCM-basa<em&gt; Drosophila melanogaster</em

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

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

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

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

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

Una manera sencilla de medir alteraciones en el comportamiento de búsqueda de recompensa Uso<em&gt; Drosophila melanogaster</em

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

Measuring and Altering Mating Drive in Male Drosophila melanogaster

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

Measuring and Altering Mating Drive in Male Drosophila melanogaster

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

La medición y la modificación de acoplamiento de unidad en Male<em&gt; Drosophila melanogaster</em

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

Drosophila Courtship Conditioning As a Measure of Learning and Memory

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

Drosophila Courtship Conditioning As a Measure of Learning and Memory

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

Drosophila condicionamiento del corte como una medida de aprendizaje y memoria

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

Un método para probar el efecto de las señales ambientales sobre el comportamiento de apareamiento en

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