Name: taste preference assay for adult drosophila
Uploaded: 2016-09-08T12:30:00.000+00:00
Duration: 4 min 31 s
Description: Watch this Scientific Journal Video about Taste Preference Assay for Adult Drosophila at JoVE.com

We would like to thank members of the Tessier lab for critical reading of this manuscript and helpful suggestions during the preparation of this protocol.

1. fame <ol><li> Preparare volare fiale fame saturando un batuffolo di cotone con 18,2 MW acqua sul fondo di una fiala mosca standard. In alternativa, allo stesso modo di saturare una piccola striscia di carta da filtro con 18,2 MW acqua e posto in un angolo all&#39;interno della fiala. </li><li> Raccogliere vola in gruppi di circa 100 animali su un rilievo di CO 2 e quindi aggiungere le mosche per una fiala preparato. Nota: i migliori risultati si ottengono con animali che sono meno di 5 giorni. Tuttavia, l&#39;esatta età degli animali può essere controllata come variabile sperimentale per determinare le variazioni di preferenza di gusto nel tempo. </li><li> Utilizzare un batuffolo di cotone o schiuma tappo per fissare le fiale chiuse. Posizionare le fiale dalla loro parte in un incubatore ambiente controllato. Mantenere la temperatura a 25 ° C, e l&#39;umidità superiore al 70%. Lascia fiale intatte per 24 ore. </li></ol> 2. Taste Preferenza Assay <ol><li> Preparare tutti tastants per il saggio lo stesso giorno uns test. Nota: I tastants esatta da utilizzare varia a seconda domanda sperimentale viene chiesto. I seguenti sono esempi di tastants utilizzati in questo protocollo. Vedere la sezione 4 per ottimizzazioni. <ol><li> Preparare il controllo tastant (1 mM saccarosio) mediante la combinazione di 10 ml di 100 mm soluzione di saccarosio, 13 ml di colorante alimentare rosso, e 977 ml di 18,2 MW acqua. </li><li> Preparare tastant sperimentale (5 mm saccarosio) mediante la combinazione di 50 ml di 100 mm soluzione di saccarosio, 10 ml di colorante alimentare blu, e 940 ml di 18,2 MW acqua. </li></ol></li><li> Rendere camere dosaggio usando da 100 mm x 15 mm piatto di plastica standard di petri preparato nel seguente modo: <ol><li> Mettere tre gocce 10 microlitri di controllo tastant più vicino al bordo del piatto a ore 12 e un altro 3 gocce a ore 6. Assicurarsi che la distanza tra gocce è simile. </li><li> Inserire tre gocce 10 microlitri di tastant sperimentale vicina al bordo della piastra a ore 3 e unaltri 3 gocce alle ore 9. Assicurarsi che la distanza tra gocce è simile. </li><li> Ripetere i punti 2.2.1 e 2.2.2 per il numero di repliche, se lo desideri. </li></ol></li><li> Vuoto 1 fiala di ~ 100 mosche di fame su un tappetino di CO 2 giusto il tempo per anestetizzare gli animali (circa 10 sec). Spennellate gli animali in mezzo a una camera di test preparata e coprire con il coperchio piatto. Nota: più lunghi periodi di esposizione CO 2 dovrebbe essere evitato per migliorare i tempi di recupero e l&#39;interferenza con il limite di comportamento alimentare. L&#39;esposizione al ghiaccio (~ 5 min) può essere utilizzato per anestetizzare per evitare di CO 2 effetti comportamentali che possono derivare da un&#39;esposizione limitata. </li><li> Posizionare la camera di test in una scatola di cartone opaco. Assicurarsi di etichettare l&#39;esterno della scatola con la condizione e il genotipo in fase di test. </li><li> Posizionare l&#39;intera impostazione (camera di analisi contenuta in scatola di cartone a partire dal punto 2.4) in un incubatore a 25 ° con almeno il 70% di umidità per 2 ore. </li&gt; <li> Ripetere i passaggi 2.3 tramite 2.5 per tutte le repliche. </li><li> Dopo 2 ore, posizionare le camere di test, ancora contenuti in scatole di cartone, direttamente in un congelatore a -20 ° C fino al momento per la quantificazione. </li></ol> 3. Gusto Preferenza Assay Quantificazione <ol><li> Lasciare una sola camera di test per riscaldare a temperatura ambiente (circa 5 min). </li><li> Sotto un microscopio dissezione, utilizzando un pennello o un paio di pinze, animali di gruppo in base al colore della loro addome: rosso, blu, viola o trasparente (Figura 1). </li><li> Registrare il numero di animali in ciascun gruppo. Considerare gli animali chiare per non hanno partecipato al test e quindi non includerli in tutti i calcoli. </li><li> Calcolare l&#39;indice preferenza, secondo una delle seguenti formule: <ol><li> Se il tastant sperimentale di interesse viene aggiunto il colorante rosso, quindi utilizzare (N + rosso 0,5N viola) / (N + N rosso N purpl blue +e). </li><li> Se il tastant sperimentale viene aggiunto il colorante blu, quindi regolare l&#39;equazione (N blu + 0,5 N viola) / (N + N blu rosso + N viola). </li></ol></li><li> Ripetere i calcoli per tutte le condizioni sperimentali e replicati. </li></ol> 4. Ottimizzazione del Gusto Preferenza Assay <ol><li> Empiricamente determinare la concentrazione di indicatori colorante alimentare per essere utilizzato in modo colorante alimentare non influenza l&#39;esito del test gusto, come segue: <ol><li> Preparare 4 tastants utilizzando la stessa mescola di base (ad esempio, 5 mM saccarosio), come indicato al punto 2.1, ma omettere il colorante alimentare. </li><li> Aggiungere 1,3% colorante alimentare rosso per una delle sostanze sapide. Effettuare le restanti 3 tastants con prodotti alimentari blu coloranti di concentrazioni variabili in ogni tubo (ad esempio 0,6%, 1% e 1,3%). </li><li> protocollo completo passi 2.2 attraverso 3.4 per ogni coppia tastant: 1,3% vs. 0,6% rosso blu; 1,3% rispetto al rosso 1% e il blu1,3% contro 1,3% rosso blu. </li><li> Ripetere il passaggio 4.1.1-4.1.3 con diverse percentuali di colorante alimentare blu fino alle medie indice preferenza un valore pari a 0 (Figura 2). Nota: Come punto di partenza, 1,3% colorante alimentare rosso accoppiato con 1% colorante alimentare blu produce in genere buoni risultati. Se nessuna concentrazione soddisfacente di colorante alimentare blu può essere abbinato al 1,3% di colorante, poi passo 4.1.1 tramite 4.1.3 può essere ripetuta con concentrazioni variabili di colorazione rossa e una costante concentrazione di colorante alimentare blu. </li><li> Analizzare tutte le condizioni per essere testati con le stesse concentrazioni di colorante alimentare ottimizzati. </li></ol></li></ol>

<ol>
	<li>Herrero, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fruit+fly+behavior+in+response+to+chemosensory+signals.">Fruit fly behavior in response to chemosensory signals.</a> Peptides. 38 (2), 228-237 (2012).</li><li>Vosshall, L. B., Stocker, R. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+architecture+of+smell+and+taste+in+Drosophila.">Molecular architecture of smell and taste in Drosophila.</a> Annu Rev Neurosci. 30, 505-533 (2007).</li><li>Harris, D. T., Kallman, B. R., Mullaney, B. C., Scott, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Representations+of+Taste+Modality+in+the+Drosophila+Brain.">Representations of Taste Modality in the Drosophila Brain.</a> Neuron. 86 (6), 1449-1460 (2015).</li><li>Hong, E. J., Wilson, R. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simultaneous+encoding+of+odors+by+channels+with+diverse+sensitivity+to+inhibition.">Simultaneous encoding of odors by channels with diverse sensitivity to inhibition.</a> Neuron. 85 (3), 573-589 (2015).</li><li>Kain, P., Dahanukar, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Secondary+taste+neurons+that+convey+sweet+taste+and+starvation+in+the+Drosophila+brain.">Secondary taste neurons that convey sweet taste and starvation in the Drosophila brain.</a> Neuron. 85 (4), 819-832 (2015).</li><li>Masek, P., Worden, K., Aso, Y., Rubin, G. M., Keene, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+dopamine-modulated+neural+circuit+regulating+aversive+taste+memory+in+Drosophila.">A dopamine-modulated neural circuit regulating aversive taste memory in Drosophila.</a> Curr Biol. 25 (11), 1535-1541 (2015).</li><li>Charlu, S., Wisotsky, Z., Medina, A., Dahanukar, A. <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+sweet+and+bitter+taste+neurons+in+Drosophila+melanogaster.">Acid sensing by sweet and bitter taste neurons in Drosophila melanogaster.</a> Nat Commun. 4, 2042 (2013).</li><li>Clyne, P. J., Warr, C. G., Carlson, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Candidate+taste+receptors+in+Drosophila.">Candidate taste receptors in Drosophila.</a> Science. 287 (5459), 1830-1834 (2000).</li><li>Scott, K., 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=A+chemosensory+gene+family+encoding+candidate+gustatory+and+olfactory+receptors+in+Drosophila.">A chemosensory gene family encoding candidate gustatory and olfactory receptors in Drosophila.</a> Cell. 104 (5), 661-673 (2001).</li><li>Kim, S. H., 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=Drosophila+TRPA1+channel+mediates+chemical+avoidance+in+gustatory+receptor+neurons.">Drosophila TRPA1 channel mediates chemical avoidance in gustatory receptor neurons.</a> Proc Natl Acad Sci U S A. 107 (18), 8440-8445 (2010).</li><li>Koh, T. W., 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=The+Drosophila+IR20a+clade+of+ionotropic+receptors+are+candidate+taste+and+pheromone+receptors.">The Drosophila IR20a clade of ionotropic receptors are candidate taste and pheromone receptors.</a> Neuron. 83 (4), 850-865 (2014).</li><li>Zhang, Y. V., Ni, J., Montell, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+molecular+basis+for+attractive+salt-taste+coding+in+Drosophila.">The molecular basis for attractive salt-taste coding in Drosophila.</a> Science. 340 (6138), 1334-1338 (2013).</li><li>Zhang, Y. V., Raghuwanshi, R. P., Shen, W. L., Montell, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Food+experience-induced+taste+desensitization+modulated+by+the+Drosophila+TRPL+channel.">Food experience-induced taste desensitization modulated by the Drosophila TRPL channel.</a> Nat Neurosci. 16 (10), 1468-1476 (2013).</li><li>Liman, E. R., Zhang, Y. V., Montell, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Peripheral+coding+of+taste.">Peripheral coding of taste.</a> Neuron. 81 (5), 984-1000 (2014).</li><li>Rodrigues, V., Cheah, P. Y., Ray, K., Chia, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=malvolio,+the+Drosophila+homologue+of+mouse+NRAMP-1+(Bcg),+is+expressed+in+macrophages+and+in+the+nervous+system+and+is+required+for+normal+taste+behaviour.">malvolio, the Drosophila homologue of mouse NRAMP-1 (Bcg), is expressed in macrophages and in the nervous system and is required for normal taste behaviour.</a> EMBO J. 14 (13), 3007-3020 (1995).</li><li>Tanimura, T., Isono, K., Yamamoto, M. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Taste+sensitivity+to+trehalose+and+its+alteration+by+gene+dosage+in+Drosophila+melanogaster.">Taste sensitivity to trehalose and its alteration by gene dosage in Drosophila melanogaster.</a> Genetics. 119 (2), 399-406 (1988).</li><li>Weiss, L. A., Dahanukar, A., Kwon, J. Y., Banerjee, D., Carlson, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+molecular+and+cellular+basis+of+bitter+taste+in+Drosophila.">The molecular and cellular basis of bitter taste in Drosophila.</a> Neuron. 69 (2), 258-272 (2011).</li><li>French, A. S., 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=Dual+mechanism+for+bitter+avoidance+in+Drosophila.">Dual mechanism for bitter avoidance in Drosophila.</a> J Neurosci. 35 (9), 3990-4004 (2015).</li><li>Deshpande, S. A., 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=Quantifying+Drosophila+food+intake:+comparative+analysis+of+current+methodology.">Quantifying Drosophila food intake: comparative analysis of current methodology.</a> Nat Methods. 11 (5), 535-540 (2014).</li></ol>

The authors declare that they have no competing financial interests.

Abbiamo descritto un protocollo semplice ma efficace per determinare preferenza di gusto in Drosophila. Le versioni di questo test sono abitualmente utilizzati in esperimenti per determinare i contributi di recettori gustativi (GRS) a percepire le diverse qualità (amaro, dolce, acido, salato e umami) di composti di gusto. Il genoma di Drosophila contiene circa 60 geni che codificano 68 individuati recettori gustativi da splicing alternativo 8,9. Tuttavia, altre proteine ​​come recettori...

Gli animali usano chemosensation distinguere condizioni vantaggiose a parte condizioni svantaggiose. Questa percezione può essere fondamentale per cose come determinare la migliore fonte di cibo, evitando sostanze tossiche o determinare il miglior partner di accoppiamento 1. Chemosensation è spesso diviso in due componenti sensoriali: olfatto e sensi gustative. Una principale caratteristica distintiva di questi sensi è che l&#39;olfatto (odore) è usato per campionare l&#39;ambiente chimico gassoso circostante mentre gustation (gusto) richiede il contatto fisico con un substrato non volatile. Entrambe le modalità sensoriali stimolano risposte neurologiche che vengono elaborati e decodificati nel cervello per produrre l&#39;attrattiva o repulsiva comportamento appropriato 2. Questi sensi sono quindi fondamentale per la sopravvivenza degli animali. La mosca della frutta Drosophila melanogaster è un organismo modello che continua a crescere in popolarità per l&#39;uso in capireing come gli insetti percepiscono odore e sapore. Frutta mosche offrono enormi vantaggi rispetto ad altri sistemi modello per la ricchezza di strumenti genetici disponibili per la dissezione di percorsi molecolari, cellulari e comportamentali. Il lavoro nel corso degli ultimi 15 anni è stato particolarmente determinante nel caratterizzare le identità cellulari specifici, recettori neuronali, e meccanismi coinvolti sia in odore e il sapore di segnalazione. Ora, il potere della genetica della Drosophila viene usata per chiarire ulteriormente come questi processi sono codificati al singolo neurone e il circuito unico livello 3-6. Pertanto, le analisi che forniscono facilmente lanciati letture di alterazioni percorsi sensoriali sono di vitale importanza per il continuo avanzamento di questi campi. Mentre molto si sa su come i segnali olfattivi sono codificati ed elaborati nel cervello, e tanto meno è capito su meccanismi simili nel percorso gustativo. Descriviamo qui un protocollo che può essere utilizzato per verificare preferen gustoce in Drosophila. Drosophila, come i mammiferi, in genere preferiscono i composti di degustazione di dolci al contrario di composti sapore amaro. Qualsiasi combinazione di queste fonti di cibo può essere utilizzato in questo progetto sperimentale per determinare come note alterazioni genetiche influenzano la scelta gusto. Inoltre, le strategie di intervento farmacologico può analogamente essere valutati per i loro effetti sulla preferenza di gusto degli animali. La facilità e la flessibilità di questo test lo rende un paradigma utile per comprendere la natura della percezione gustativa in Drosophila.

<table><tbody><tr><td>Blue Food Coloring (Water, Propylene Glycol, FD&amp;amp;C Blue 1 and Red 40, Propylparaben)</td><td>McCormick</td><td>N/A</td><td></td></tr><tr><td>Cryo/Freezer Boxes w/o Dividers</td><td>Fisher</td><td>03-395-455</td><td></td></tr><tr><td>Dumont #5 Forceps</td><td>Fine Science Tools</td><td>11251-20</td><td></td></tr><tr><td>Glacial Acetic Acid</td><td>Fisher</td><td>BP2401-500</td><td></td></tr><tr><td>Leica S6 E Stereozoom 0.63X-4.0X microscope</td><td>W. Nuhsbaum, Inc.</td><td>10446294</td><td></td></tr><tr><td>Petri dish (100 mm x 15 mm)</td><td>BD Falcon</td><td>351029</td><td>Reuseable if thoroughly washed and dried</td></tr><tr><td>Quick-Snap Microtubes</td><td>Alkali Scientific Inc.</td><td>C3017</td><td></td></tr><tr><td>Red Food Coloring (Water, Propylene Glycol, FD&amp;amp;C Reds 40 and 3, Propylparaben)</td><td>McCormick</td><td>N/A</td><td></td></tr><tr><td>Sucrose</td><td>IBI Scientific</td><td>IB37160</td><td></td></tr></tbody></table>

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.

Alcuni risultati tipici da saggi di preferenza di gusto sono riportati di seguito. Nella maggior parte degli esperimenti qualche variazione di intensità di colorazione addominale sarà visto (Figura 1). Qualsiasi colorazione nell&#39;addome se intensa o debole è considerato un ingestione positivo. E &#39;quindi consigliabile per i ricercatori a segnare animali, mentre ciechi alla condizione sperimentale in modo da limitare i potenziali errori. <p class="jove_conten...

Watch this Scientific Journal Video about Taste Preference Assay for Adult Drosophila at JoVE.com

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.

Gusto Preferenza Assay per adulti<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 ...

taste-preference-assay-for-adult-drosophila

Research

JoVE Journal

Neuroscience

Taste Preference Assay for Adult Drosophila

9.6K Views.  Indiana University School of Medicine-South Bend. 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.

Video: Taste Preference Assay for Adult Drosophila

Appetitive Associative Olfactory Learning in Drosophila Larvae

In the following we describe the methodological details of appetitive associative olfactory learning in Drosophila larvae. The setup, in combination with genetic interference, provides a handle to analyze the neuronal and molecular fundamentals of specifically associative learning in a simple larval brain.
Organisms can use past experience to adjust present behavior. Such acquisition of behavioral potential can be defined as learning, and the physical bases of these potentials as memory traces1-4. Neuroscientists try to understand how these processes are organized in terms of molecular and neuronal changes in the brain by using a variety of methods in model organisms ranging from insects to vertebrates5,6. For such endeavors it is helpful to use model systems that are simple and experimentally accessible. The Drosophila larva has turned out to satisfy these demands based on the availability of robust behavioral assays, the existence of a variety of transgenic techniques and the elementary organization of the nervous system comprising only about 10,000 neurons (albeit with some concessions: cognitive limitations, few behavioral options, and richness of experience questionable)7-10.
Drosophila larvae can form associations between odors and appetitive gustatory reinforcement like sugar11-14. In a standard assay, established in the lab of B. Gerber, animals receive a two-odor reciprocal training: A first group of larvae is exposed to an odor A together with a gustatory reinforcer (sugar reward) and is subsequently exposed to an odor B without reinforcement 9. Meanwhile a second group of larvae receives reciprocal training while experiencing odor A without reinforcement and subsequently being exposed to odor B with reinforcement (sugar reward). In the following both groups are tested for their preference between the two odors. Relatively higher preferences for the rewarded odor reflect associative learning - presented as a performance index (PI). The conclusion regarding the associative nature of the performance index is compelling, because apart from the contingency between odors and tastants, other parameters, such as odor and reward exposure, passage of time and handling do not differ between the two groups9.

Appetitive Associative Olfactory Learning in Drosophila Larvae

Drosophila larvae are able to associate odor stimuli with gustatory reward. Here we describe a simple behavioral paradigm that allows the analysis of appetitive associative olfactory learning.

Appetitivo apprendimento associativo olfattiva in<em&gt; Drosophila</em&gt; Le larve

In the following we describe the methodological  details of appetitive associative olfactory learning in Drosophila larvae. The setup, in combination with ...

Ricerca

Neuroscienze

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.

A Single-fly saggio per comportamento alimentare 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 ...

Testing Drosophila Olfaction with a Y-maze Assay

Detecting signals from the environment is essential for animals to ensure their survival. To this aim, they use environmental cues such as vision, mechanoreception, hearing, and chemoperception through taste, via direct contact or through olfaction, which represents the response to a volatile molecule acting at longer range. Volatile chemical molecules are very important signals for most animals in the detection of danger, a source of food, or to communicate between individuals. Drosophila melanogaster is one of the most common biological models for scientists to explore the cellular and molecular basis of olfaction. In order to highlight olfactory abilities of this small insect, we describe a modified choice protocol based on the Y-maze test classically used with mice. Data obtained with Y-mazes give valuable information to better understand how animals deal with their perpetually changing environment. We introduce a step-by-step protocol to study the impact of odorants on fly exploratory response using this Y-maze assay.

Testing Drosophila Olfaction with a Y-maze Assay

We present a choice test to reveal the influence of odorants on Drosophila behavior using a Y-maze assay.

Test<em&gt; Drosophila</em&gt; Olfatto con un test Y-maze

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

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

Recently, egg-laying preference of Drosophila has emerged as a genetically tractable model to study the neural basis of simple decision-making processes. When selecting sites to deposit their eggs, female flies are capable of ranking the relative attractiveness of their options and choosing the &#34;greater of two goods.&#34; However, most egg-laying preference assays are not practical if one wants to take a systematic genetic screening approach to search for the circuit basis underlying this simple decision-making process, as they are population-based and laborious to set up. To increase the throughput of studying of egg-laying preferences of single females, we developed custom chambers that each can simultaneously assay egg-laying preferences of up to thirty individual flies as well as a protocol that ensures each female has a high egg-laying rate (so that their preference is readily discernable and more convincing). Our approach is simple to execute and produces very consistent results. Additionally, these chambers can be equipped with different attachments to allow video recording the egg-laying animals and to deliver light for optogenetics studies. This article provides the blueprints for fabricating these chambers and the procedure for preparing the flies to be assayed in these chambers.

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

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.

High Throughput test per esaminare Preferenze deposizione delle uova dei singoli<em&gt; Drosophila melanogaster</em

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

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

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

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

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

Un modo semplice per misurare alterazioni in ricompensa il comportamento di ricerca Utilizzando<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 ...

The CApillary FEeder Assay Measures Food Intake in Drosophila melanogaster

For most animals, feeding is an essential behavior for securing survival, and it influences development, locomotion, health and reproduction. Ingestion of the right type and quantity of food therefore has a major influence on quality of life. Research on feeding behavior focuses on the underlying processes that ensure actual feeding and unravels the role of factors regulating internal energy homeostasis and the neuronal bases of decision-making. The model organism Drosophila melanogaster, with its great variety of genetically traceable tools for labeling and manipulating single neurons, allows mapping of neuronal networks and identification of molecular signaling cascades involved in the regulation of food intake. This report demonstrates the CApillary FEeder assay (CAFE) and shows how to measure food intake in a group of flies for time spans ranging from hours to days. This easy-to-use assay consists of glass capillaries filled with liquid food that flies can freely access and feed on. Food consumption in the assay is accurately determined using simple measurement tools. Herein we describe step-by-step the method from setup to successful execution of the CAFE assay, and provide practical examples to analyze the food intake of a group of flies under controlled conditions. The reader is guided through possible limitations of the assay, and advantages and disadvantages of the method compared to other feeding assays in D. melanogaster are evaluated.

The CApillary FEeder Assay Measures Food Intake in Drosophila melanogaster

The CApillary FEeder (CAFE) assay is a simple, budget-friendly, highly reliable method for investigating mechanisms underlying food intake. Used with the highly versatile genetic model organism Drosophila melanogaster, it provides a powerful means of gaining new insights into regulatory mechanisms of food intake.

Il capillare alimentatore Assay Misure assunzione di cibo in<em&gt; Drosophila melanogaster</em

For most animals, feeding is an essential behavior for securing survival, and it influences development, locomotion, health and reproduction. Ingestion of ...

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.

Un saggio rapido di preferenza alimentare 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 ...

Feeding Test: Assessment of Feeding Preferences in Drosophila melanogaster

Source: Bovier, T. F., et al. <a href="https://www.jove.com/video/59535/" target="_blank">Methods to Test Endocrine Disruption in Drosophila melanogaster</a>. J. Vis. Exp. (2019).
This video describes a feeding assay for fruit flies in which colored food is used to monitor feeding and the effect added compounds may have on feeding activity. The example protocol tests the effects of several endocrine disrupting chemicals (EDC).

Test di alimentazione: Valutazione delle preferenze alimentari in Drosophila melanogaster

Source: Bovier, T. F., et al. Methods to Test Endocrine Disruption in Drosophila melanogaster. J. Vis. Exp. (2019).
This video describes a feeding assay ...

JoVE Encyclopedia of Experiments

Biologia

Drosophila melanogaster (fruit fly)

Adult

Temperature Gradient Assay: A Method to Test Temperature Preference in Drosophila Larvae

Source: Liu, J., et al. <a href="https://www.jove.com/video/57963/" target="_blank">A Temperature Gradient Assay to Determine Thermal Preferences of Drosophila Larvae</a>. J. Vis. Exp. (2018).
Drosophila fruit flies can distinguish small changes in temperature and therefore select environments with favorable thermal conditions. This video describes a behavioral assay that tests the temperature preference of Drosophila larvae on a linear thermal gradient.

Saggio del gradiente di temperatura: un metodo per testare la preferenza di temperatura nelle larve di Drosophila

Source: Liu, J., et al. A Temperature Gradient Assay to Determine Thermal Preferences of Drosophila Larvae. J. Vis. Exp. (2018).
Drosophila fruit flies ...

Larva

Taste Preference Assay: A Method for Measuring Feeding Behavior in Drosophila

Source: Bantel, A. P. and Tessier, C. R. <a href="https://www.jove.com/video/54403/" target="_blank">Taste Preference Assay for Adult Drosophila</a>. J. Vis. Exp. (2016).
This video describes the taste preference assay, a behavioral method used to measure attraction or avoidance towards colored solutions that taste differently by assessing the fly&#39;s abdominal coloration after ingestion of the preferred substance. The featured protocol demonstrates the procedure used to measure flies&#39; preference towards solutions of varying sucrose concentrations.

Saggio delle preferenze gustative: un metodo per misurare il comportamento alimentare in Drosophila

Source: Bantel, A. P. and Tessier, C. R. Taste Preference Assay for Adult Drosophila. J. Vis. Exp. (2016).
This video describes the taste preference ...

Gusto Preferenza Assay per adulti

Riepilogo

Esplora Altri Video

Capitoli in questo video

Appetitivo apprendimento associativo olfattiva in

A Single-fly saggio per comportamento alimentare in

Test

High Throughput test per esaminare Preferenze deposizione delle uova dei singoli

Un modo semplice per misurare alterazioni in ricompensa il comportamento di ricerca Utilizzando

Il capillare alimentatore Assay Misure assunzione di cibo in

Un saggio rapido di preferenza alimentare in Drosophila

Test di alimentazione: Valutazione delle preferenze alimentari in Drosophila melanogaster

Saggio del gradiente di temperatura: un metodo per testare la preferenza di temperatura nelle larve di Drosophila

Saggio delle preferenze gustative: un metodo per misurare il comportamento alimentare in Drosophila