We thank the Duke Physics Shop, especially Phil Lewis, for helping us build the behavioral apparatus and attachments and creating the drawing. This work is funded by the National Institutes of Health under award number R01GM100027.

1. Flies Vorbereitung zu test <ol><li> Kultur fliegt auf Standard-Melasse / cornmeal Medien bei Inkubator bei 25 ° C und 65% Luftfeuchtigkeit. Achten Sie darauf, nicht die Fläschchen überfüllen. Zum Beispiel setzen 8 Frauen und 6 Männer in einen schmalen Essen Fläschchen. Hinweis: Die &quot;narrow Essen Fläschchen&quot; verwendet hier einen Innendurchmesser von 2,3 cm. Wir typischerweise etwa 10 ml fly Nahrung in jedem Fläschchen abgefüllt. Die Fliege Essen Rezept wir verwendet wird hier beschrieben: http://flystocks.bio.indiana.edu/Fly_Work/media-recipes/molassesfood.html. </li><li> 2 - 3 Tage nach dem Schlüpfen Weibchen, bereiten Fläschchen mit frischer Hefe - Paste in 1E , wie dargestellt. Machen Frischhefe Paste durch 6 g aktiver Hefe-Mischung mit 10 ml von 0,5% Propionsäure. Verwenden Sie einen Spachtel, um den Hefe-Paste auf die Seitenwand der Lebensmittel Fläschchen anzuwenden. Sammeln Sie 30 - 35 geschlüpften Weibchen zusammen mit 20 bis 25 Männer in die Fläschchen. <ol><li> Im Allgemeinen sammeln die Männchen von den gleichen Fläschchen, wo die Weibchen gesammelt werden(So ​​wie die Zeit und Mühe zu sparen). Wenn es jedoch Besorgnis über die Fruchtbarkeit von Männern ist, verwenden WT männlichen statt. Auch Hefe-Paste ist wichtig für die Stimulierung der Eierproduktion ist es daher bereiten täglich frisch. </li></ol></li><li> Aufrechterhaltung der gesammelten Weibchen / Männchen bei 25 ° C, 65% Luftfeuchtigkeit, es sei denn, die Fliegen Temperatur empfindlich sind. </li><li> Nach ca. 4 - 5 Tagen überprüfen Sie die Fläschchen zu sehen, ob Frauen bereit für Eiablage Experimente sind. Sie sind bereit , wenn die Oberfläche des Nahrungsmittel Medium nass von Larven ist aktiv in der Lebensmittel gräbt (siehe Abbildung 1 E - F). Hinweis: Frauen vorenthalten Regel Eier zu legen, wenn die Nahrungsmitteloberfläche nass wird und kriecht mit Larven. Dieser Schritt stellt sicher , dass Frauen bereit sein würde , viele Eier zu legen , wenn sie Substrate mit der gewünschten Textur auftreten (z. B. ~ 1% Agarose). </li></ol> 2. Kammer Konstruktion, Montage, Test-Setup <ol><li> Haben Sie einen Maschinenhalle bauen Acryl Eiablage Kammern (FiAbbildung 1A - D). Die technische Zeichnung der verschiedenen Stücke wurden in Ergänzungs gezeigt . 1 - 3 Bilder mit hoher Auflösung können auch hier bestellt werden (http://www.rebeccayang.org/pdf/chamber%20design.pdf). </li><li> Kunststoffplatten in die Lade (oben) Stück der Kammer ein , wie in 1C gezeigt. Dies dient als Bodenfläche während des Ladens in einzelne Eiablage Arenen fliegt. </li><li> Anesthetize Weibchen auf einem CO 2 Pad und laden Sie sie einzeln in jedem Eiablage Arena. Lassen Sie ~ 30 min für Fliegen aus dem CO 2 zu erholen und an die neue Umgebung gewöhnt zu werden. </li><li> Bereiten Sie die Agarose-Substrate. <ol><li> Der Einfachheit halber halten eine premade Flasche geschmolzen 1% Agarose in einem 55 C-Wasserbad °. </li><li> Fügen Sie die gewünschte Menge an Lager Saccharose-Lösung (2 M) in eine 50 ml konischen Röhrchen und mit Hilfe einer entsprechenden Menge an Agarose gemischt werden. Um zum Beispiel die 150 mM Saccharose Substrat, pla vorzubereitence 750 ul 2 M Saccharose-Lösung in das Röhrchen und füllen Sie dann das Rohr mit Agarose zur 10 ml-Markierung. </li><li> Bereiten Sie die Ebene Substrat auf die gleiche Weise, aber mit destilliertem Wasser anstelle von Saccharose-Lösung. Anmerkung: Die Endkonzentration an Agarose in diesem Protokoll als 1% etwas weniger ist. Nach unserer Erfahrung ist die genaue Konzentration der Agarose nicht so lange egal, wie es innerhalb von ~ 0,9 geregelt wird - 1,1%, und dass die beiden Substrate sind von der gleichen Agarose-Konzentration. </li></ol></li><li> Nehmen Sie das Substrat (Boden) Stück der Kammer und Pipetten 1000 ul Agarose Substrat in jeder Wanne , wie in Figur 1D zu sehen. </li><li> Lassen Sie Agarose für ~ 30 min zu verfestigen. </li><li> Nachdem die Agarose-Substrate und Fliegen bereit sind, montieren alle drei Teile des Eiablagekammer und dann die Kunststoffplatten herausnehmen. </li><li> Legen Sie die Kammern in der Fliege Inkubatoren. Anmerkung: Die Länge des Eiablage Experimenten variieren kann auf expe jerimental Bedürfnisse. Wir laufen in der Regel das Experiment O / N (14 bis 16 h). Auch keine wesentlichen Auswirkungen der zirkadianen Timing auf Eiablage Präferenzen beobachtet. </li><li> Anästhesieren Weibchen von CO 2 in die Kammer eingespritzt wird . Zerlegen Sie die Kammer, werfen Sie die betäubten Fliegen in Fliegen Leichenschauhaus (dh., Ein leerer Kaffee kann mit etwas Maisöl gefüllt). Machen Sie Fotos von den Ergebnissen für Recordkeeping (siehe Abbildung 2). </li><li> Zähle die Anzahl der Eier manuell und berechnen Vorzugs Indizes für die Analyse. Berechnen Präferenz Index wie (N a - N b) / (N + a N b) , wobei N a und N b repräsentieren Anzahl der Eier vor Ort gegen Seite b verbunden. </li></ol>

<ol>
	<li>Perisse, E., Burke, C., Huetteroth, W., Waddell, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Shocking+revelations+and+saccharin+sweetness+in+the+study+of+Drosophila+olfactory+memory.">Shocking revelations and saccharin sweetness in the study of Drosophila olfactory memory.</a> Curr. Biol. 23 (17), 752-763 (2013).</li><li>Yamamoto, N. C., Koganezawa, M. <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+circuits+of+courtship+behaviour+in+Drosophila+males.">Genes and circuits of courtship behaviour in Drosophila males.</a> Nat. Rev. Neurosci. 14 (10), 681-692 (2013).</li><li>Zwarts, L., Versteven, M., Callaerts, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetics+and+neurobiology+of+aggression+in+Drosophila.">Genetics and neurobiology of aggression in Drosophila.</a> Fly. 6 (1), 35-48 (2012).</li><li>Joseph, R. M., Devineni, A. V., King, I. F., Heberlein, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oviposition+preference+for+and+positional+avoidance+of+acetic+acid+provide+a+model+for+competing+behavioral+drives+in+Drosophila.">Oviposition preference for and positional avoidance of acetic acid provide a model for competing behavioral drives in Drosophila.</a> Proc. Natl. Acad. Sci. 106 (27), 11352-11357 (2009).</li><li>Miller, P. M., Saltz, J. B., Cochrane, V. A., Marcinkowski, C. M., Mobin, R., Turner, T. 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N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drosophila+egg-laying+site+selection+a+system+to+study+simple+decision-making+processes.">Drosophila egg-laying site selection a system to study simple decision-making processes.</a> Science. 319 (5870), 1679-1683 (2008).</li><li>Kacsoh, B. Z., Lynch, Z. R., Mortimer, N. T., Schlenke, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fruit+flies+medicate+offspring+after+seeing+parasites.">Fruit flies medicate offspring after seeing parasites.</a> Science. 339 (6122), 947-950 (2013).</li><li>Wu, C. -. L., Fu, T. -. F., Chou, Y. -. Y., Yeh, S. -. 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H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanosensitive+neurons+on+the+internal+reproductive+tract+contribute+to+egg-laying+induced+acetic+acid+attraction+in+Drosophila.">Mechanosensitive neurons on the internal reproductive tract contribute to egg-laying induced acetic acid attraction in Drosophila.</a> Cell Rep. 9 (2), 522-530 (2014).</li><li>Stern, U., Zhu, E. Y., He, R., Yang, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-duration+animal+tracking+in+difficult+lighting+conditions.">Long-duration animal tracking in difficult lighting conditions.</a> Sci. Rep. 5, 10432 (2015).</li><li>Zhu, E. Y., Guntur, A. R., He, R., Stern, U., Yang, C. 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The authors have nothing to disclose.

Die Kammern und Protokolle hier beschrieben haben mehrere Verbesserungen gegenüber früheren Eiablage Assays. Erstens erhöhen sie den Durchsatz erheblich Präferenzen der einzelnen Tiere Untersuchung. Jede Kammer kann 30 einzelne Frauen-Assay und es dauert weniger als eine Stunde einzurichten. Zweitens erhöhen sie die Konsistenz der Eiablage Präferenzen gegenüber früheren Methoden. Die Standardisierung der Abmessungen der Arena, Größe der Eiablage Substrate und Abstand zwischen den Substraten macht es leichter, ...

Drosophila melanogaster is a powerful genetic model organism to study the neural basis of behaviors. The rapid developments of genetic tools to manipulate neurons in a targeted manner and the emergence of sophisticated behavioral analysis tools have significantly improved our ability to dissect the circuit mechanisms that underlie the sensory-motor transformation processes of several innate and learned behaviors1-3.
Drosophila egg-laying is a suitable model to study the neural basis of simple decision making processes. In particular, Drosophila females have been shown to possess the ability to compare and rank their options before "committing" to depositing an egg onto a given option4-8. For example, when given only a plain (sucrose-free) substrate or only a sucrose-containing substrate, females readily accept either option for egg-laying. However, when presented with both options, females robust reject the sucrose substrate in some contexts7,9,10. Relatively little is known about the neural mechanisms that allow females to "choose the greater of two goods", however. A major obstacle has been the lack of an efficient method to assay egg-laying preferences such that one can use a systematic genetic screening approach to study this problem.
In this report, we describe the protocol we developed that allows egg-laying preferences of females to be assayed at single-animal resolution and with substantially improved throughput and consistency over previous methods. Specifically, we provide the blueprints for constructing the chambers we designed, the protocol for preparing the females so that each is primed to lay many eggs, and the protocol for using the chambers.

<table><tbody><tr><td>UltraPure Agarose&amp;nbsp;</td><td>Invitrogen</td><td>16500-500</td><td></td></tr><tr><td>Sucrose</td><td>Sigma</td><td>S0389</td><td></td></tr><tr><td>Water bath&amp;nbsp;</td><td>Fisher</td><td>15-462-6Q</td><td></td></tr><tr><td>LifeCam Cinema webcam</td><td>Microsoft</td><td>H5D-00013</td><td></td></tr><tr><td>Red LEDs</td><td>Cree</td><td>C503B-RAN-CA0B0AA1</td><td></td></tr><tr><td>Egg-laying chambers</td><td>Custom Built</td><td></td><td></td></tr><tr><td>Camera holders</td><td>Custom Built</td><td></td><td></td></tr><tr><td>LED holders</td><td>Custom Built</td><td></td><td></td></tr><tr><td>Fly vials (narrow)</td><td>Genesee</td><td>32-116BC</td><td></td></tr></tbody></table>

high throughput assay to examine egg-laying preferences of individual drosophila melanogaster

Vor kurzem Eiablage bevorzugt von Drosophila als genetisch manipulierbaren Modell entstand die neuronale Grundlage einfacher Entscheidungsprozesse zu studieren. Bei der Auswahl von Standorten um ihre Eier abzulegen, sind weibliche Fliegen der Lage, die relative Attraktivität ihrer Optionen Ranking und die Auswahl von &quot;größeren von zwei Waren.&quot; Allerdings sind die meisten Eiablage bevorzugt Assays sind nicht praktisch, wenn man will, eine systematische genetische Screening-Ansatz zu übernehmen für die Schaltung Basis zu suchen, diese einfache Entscheidungsprozess zugrunde liegen, wie sie sind populationsbasierte und arbeitsaufwendig einzurichten. Um den Durchsatz des Studiums der Eiablage Präferenzen der einzelnen Frauen zu erhöhen, entwickelten wir individuelle Kammern, die jeweils gleichzeitig Assay Eiablage Vorlieben von bis zu dreißig Einzel sowie ein Protokoll fliegt, die jedes Weibchen sorgt für eine hohe Eiablagerate (so dass ihre Präferenz ist leicht erkennbar und überzeugender). Unser Ansatz ist einfach auszuführenund produziert sehr konsistente Ergebnisse. Darüber hinaus können diese Kammern mit verschiedenen Aufsätzen ausgestattet werden, um Video die Eiablage Tiere Aufnahme zu ermöglichen und Licht für die Optogenetik Studien zu liefern. Dieser Artikel enthält die Baupläne für diese Kammern Herstellung und das Verfahren zur Herstellung der Fliegen in diesen Kammern untersucht werden.

Die Eiablage Kammern bestehen aus mehreren Teilen zusammengesetzt: ein Substrat (unten) Stück, einen Teiler (Mitte) Stück, eine Lade (oben) Stück, und 2 Schiebetüren (1A - D). Diese Stücke werden verwendet , um unabhängig Setup Fliegen und Substraten vor Eiablage Experimenten. 1F zeigt , wie Fläschchen aussehen sollte , wenn weibliche Fliegen bereit für Eiablage sind. Wenn Fliegen eine Wahl zwischen einem einfachen und einem Substrat Saccharose-h...

Watch this Scientific Journal Video about High Throughput Assay to Examine Egg-Laying Preferences of Individual Drosophila melanogaster at JoVE.com

High Throughput Assay to Examine Egg-Laying Preferences of Individual Drosophila melanogaster

Assay mit hohem Durchsatz Eiablage Präferenzen der einzelnen zu untersuchen<em&gt; Drosophila melanogaster</em

This protocol describes a high throughput assay for testing egg-laying preferences of Drosophila melanogaster at single-animal resolution. This assay provides a simple, efficient, and scalable platform to identify genes and circuit components that control a simple decision-making process.

Recently, egg-laying preference of Drosophila has emerged as a genetically tractable model to study the neural basis of simple decision-making processes. ...

high-throughput-assay-to-examine-egg-laying-preferences-individual

Research

JoVE Journal

Neuroscience

High Throughput Assay to Examine Egg-Laying Preferences of Individual Drosophila melanogaster

7.9K Aufrufe.  Duke University. This protocol describes a high throughput assay for testing egg-laying preferences of Drosophila melanogaster at single-animal resolution. This assay provides a simple, efficient, and scalable platform to identify genes and circuit components that control a simple decision-making process.

Serie: High Throughput Assay to Examine Egg-Laying Preferences of Individual 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.

Methoden zum Test Drosophila Verhalten

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

Forschung

Neurowissenschaften

Light Preference Assay to Study Innate and Circadian Regulated Photobehavior in Drosophila Larvae

Light acts as environmental signal to control animal behavior at various levels. The Drosophila larval nervous system is used as a unique model to answer basic questions on how light information is processed and shared between rapid and circadian behaviors. Drosophila larvae display a stereotypical avoidance behavior when exposed to light. To investigate light dependent behaviors comparably simple light-dark preference tests can be applied. In vertebrates and arthropods the neural pathways involved in sensing and processing visual inputs partially overlap with those processing photic circadian information. The fascinating question of how the light sensing system and the circadian system interact to keep behavioral outputs coordinated remains largely unexplored. Drosophila is an impacting biological model to approach these questions, due to a small number of neurons in the brain and the availability of genetic tools for neuronal manipulation. The presented light-dark preference assay allows the investigation of a range of visual behaviors including circadian control of phototaxis.

Light Preference Assay to Study Innate and Circadian Regulated Photobehavior in Drosophila Larvae

Here we describe a light-dark preference test for Drosophila larva. This assay provides information about innate and circadian regulation of light sensing and processing photobehavior.

Licht Vorlieben Assay zur angeborenen und Circadian geregelten Photobehavior in Studie<em&gt; Drosophila</em&gt; Larven

Light acts as environmental signal to control animal behavior at various levels. The Drosophila larval nervous system is used as a unique model to answer ...

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 ...

Assay for Blood-brain Barrier Integrity in Drosophila melanogaster

Proper nervous system development includes the formation of the blood-brain barrier, the diffusion barrier that tightly regulates access to the nervous system and protects neural tissue from toxins and pathogens. Defects in the formation of this barrier have been correlated with neuropathies, and the breakdown of this barrier has been observed in many neurodegenerative diseases. Therefore, it is critical to identify the genes that regulate the formation and maintenance of the blood-brain barrier to identify potential therapeutic targets. In order to understand the exact roles these genes play in neural development, it is necessary to assay the effects of altered gene expression on the integrity of the blood-brain barrier. Many of the molecules that function in the establishment of the blood-brain barrier have been found to be conserved across eukaryotic species, including the fruit fly, Drosophila melanogaster. Fruit flies have proven to be an excellent model system for examining the molecular mechanisms regulating nervous system development and function. This protocol describes a step-by-step procedure to assay for blood-brain barrier integrity during the embryonic and larval stages of D. melanogaster development.

Assay for Blood-brain Barrier Integrity in Drosophila melanogaster

Blood-brain barrier integrity is critical for nervous system function. In Drosophila melanogaster, the blood-brain barrier is formed by glial cells during late embryogenesis. This protocol describes methods to assay for blood-brain barrier formation and maintenance in D. melanogaster embryos and third instar larvae.

Assay für Blut - Hirn-Barriere Integrität in Drosophila melanogaster

Proper nervous system development includes the formation of the blood-brain barrier, the diffusion barrier that tightly regulates access to the nervous ...

Entwicklungsbiologie

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.

Messen und Ändern von Mating Antrieb in 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, ...

A Temperature Gradient Assay to Determine Thermal Preferences of Drosophila Larvae

Many animals, including the fruit fly, Drosophila melanogaster, are capable of discriminating minute differences in&#160;environmental temperature, which enables them to seek out their preferred thermal landscape. To define the temperature preferences of larvae over a defined linear range, we developed an assay using a temperature gradient. To establish a single-directional gradient, two aluminum blocks are connected to independent water baths, each of which controls the temperature of individual blocks. The two blocks set the lower and upper limits of the gradient. The temperature gradient is established by placing an agarose-coated aluminum plate over the two water-controlled blocks so that the plate spans the distance between them. The ends of the aluminum plate that is set on the top of the water blocks defines the minimum and maximum temperatures, and the regions in-between the two blocks form a linear temperature gradient. The gradient assay can be applied to&#160;larvae of different ages and can be used to identify mutants that exhibit phenotypes, such as those with mutations affecting genes encoding TRP channels and opsins, which are required for temperature discrimination.

A Temperature Gradient Assay to Determine Thermal Preferences of Drosophila Larvae

Here, we present a protocol to determine the preferred environmental temperature of Drosophila larvae using a continuous thermal gradient.

Ein Temperaturgradient Assay thermische Präferenzen der Drosophila Larven bestimmen

Many animals, including the fruit fly, Drosophila melanogaster, are capable of discriminating minute differences in&#160;environmental temperature, which ...

A Rapid Food-Preference Assay in Drosophila

To select food with nutritional value while avoiding the consumption of harmful agents, animals need a sophisticated and robust taste system to evaluate their food environment. The fruit fly, Drosophila melanogaster, is a genetically tractable model organism that is widely used to decipher the molecular, cellular, and neural underpinnings of food preference. To analyze fly food preference, a robust feeding method is needed. Described here is a two-choice feeding assay, which is rigorous, cost-saving, and fast. The assay is Petri-dish-based and involves the addition of two different foods supplemented with blue or red dye to the two halves of the dish. Then, ~70 prestarved, 2-4-day-old flies are placed in the dish and&#160;allowed to choose between blue and red foods in the dark for about 90 min. Examination of the abdomen of each fly is followed by the calculation of the preference index. In contrast to multiwell plates, each Petri dish takes only ~20 s to fill and saves time and effort. This feeding assay can be employed to quickly determine whether flies like or dislike a particular food.

A Rapid Food-Preference Assay in Drosophila

We present a protocol for a two-choice feeding assay for flies. This feeding assay is fast and easy to run and is suitable not only for small-scale laboratory research, but also for high-throughput behavioral screens in flies.

Ein schneller Food-Preference-Test in Drosophila

To select food with nutritional value while avoiding the consumption of harmful agents, animals need a sophisticated and robust taste system to evaluate ...

Fly Climbing Assay: A Method to Test General Locomotion in Drosophila melanogaster

Source: Gevedon, O., et al. <a href="https://www.jove.com/video/59720/in-vivo-forward-genetic-screen-to-identify-novel-neuroprotective">In Vivo Forward Genetic Screen to Identify Novel Neuroprotective Genes in Drosophila melanogaster</a>. J. Vis. Exp. (2019).
This video describes a simple method to quantify fly locomotion and shows the climbing assay performed.&#160; The fly climbing assay is a general test of motor function which relies on a negative geotaxis response.

Fly Climbing Assay: Eine Methode zum Testen der allgemeinen Fortbewegung bei Drosophila melanogaster

Source: Gevedon, O., et al. In Vivo Forward Genetic Screen to Identify Novel Neuroprotective Genes in Drosophila melanogaster. J. Vis. Exp. (2019).
This ...

JoVE Encyclopedia of Experiments

Biologie

Drosophila melanogaster (Fruchtfliege)

Adultes Tier

Egg Laying Assay: A Method to Quantify the Egg-Laying Behavior of C. elegans

Source: Lee, S. Y. and Kang, K. <a href="https://www.jove.com/video/60419/measuring-effects-bacteria-chemicals-on-intestinal-permeability">Measuring the Effect of Chemicals on the Growth and Reproduction of&#160;Caenorhabditis elegans</a>.&#160;J. Vis. Exp. (2017).
C. elegans egg-laying behavior can provide important information in developmental, reproductive, and anatomical studies.&#160; This video introduces the important steps of an egg laying assay and shows a sample experiment in which a drug is tested for reproductive toxicity.

Egg Laying Assay: Eine Methode zur Quantifizierung des Eiablageverhaltens von C. elegans

Source: Lee, S. Y. and Kang, K. Measuring the Effect of Chemicals on the Growth and Reproduction of&#160;Caenorhabditis elegans.&#160;J. Vis. Exp. 
...

Caenorhabditis elegans (Fadenwurm)

Grundlegende Methoden

Island Assay: A High-Throughput Method to Evaluate Drosophila Locomotor Behavior

Source: Eidhof, I., et al. <a href="https://www.jove.com/video/55892/" target="_blank">High-throughput Analysis of Locomotor Behavior in the Drosophila Island Assay</a>. J. Vis. Exp. (2017).
This video describes the island assay, a behavioral method used to study Drosophila locomotion. The sample protocol features a setup for the assay compatible with semi-automated image analysis.

Island Assay: Eine Hochdurchsatzmethode zur Bewertung des Fortbewegungsverhaltens von Drosophila

Source: Eidhof, I., et al. High-throughput Analysis of Locomotor Behavior in the Drosophila Island Assay. J. Vis. Exp. (2017).
This video describes the ...

Assay mit hohem Durchsatz Eiablage Präferenzen der einzelnen zu untersuchen

Zusammenfassung

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