These studies were supported by grants from the National Institutes of Health, National Institute for Alcoholism and Alcohol Abuse to M.G. (P20AA017828, R01AA020634, P50 AA022537). The authors thank Jill Bettinger for helpful discussions and Jacqueline DeLoyht for technical assistance.

1. Giorno Prima Assay <ol><li> Raccogliere mosche in fiale alimenti freschi in gruppi di 11 (sesso unico) in breve (1-5 minuti) CO 2. </li><li> Consentire mosche di recuperare O / N in fiale alimentari in uno spazio ambiente controllato (tipicamente 25 ° C, 60% umidità relativa, 12 hr luce / buio ciclo). </li><li> Preparare la soluzione di etanolo (s) diluendo puro (100%) di etanolo in purificato (≥18 MW) di acqua a concentrazione finale (s) appropriato per l&#39;esperimento previsto. Lasciare la soluzione (s) per tornare alla RT O / N. Nota: La diluizione di etanolo è esotermica. </li></ol> 2. Giornata di test <ol><li> Per ogni flacone di mosche da testare, preparare (i) un ambiente pulito, vuoto alimentare flacone; Per esempio, il test flacone, (ii) un nuovo plug acetato di cellulosa, (iii) un tappo silicone e (iv) 1 ml di soluzione di etanolo (vedi Tabella 3). </li><li> Preparare sala prove regolando la temperatura a 20-25 ° C e umidità relativa al 55-65%. </li><li> Hanno un altro lavoratoreassegnare un codice univoco a ciascun gruppo di flaconi e registrare il codice per dopo. Posizionare fiale codificati con le mosche in sala prove per ambientarsi per pochi minuti. </li><li> Etichetta fiale test vuoti per abbinare i codici di fiale vola da 2.3. </li><li> Costruire un registro test cartaceo inserendo i codici in colonne (una colonna / flaconcino) in un foglio di calcolo simile alla Tabella 1. </li><li> Utilizzo del registro di test come una guida, organizzare fiale alimentari codificati con mosche e fiale di test vuoti in corrispondenza array in sala prove. Nota: Un ragionevole numero massimo di fiale di test è 24 (cioè, 6 set di 4 flaconcini ciascuna). </li><li> Trasferimento vola da flaconcini alimentari in abbinati / etichettati fiale test vuoto e inserire immediatamente spine in acetato di cellulosa in fiale di prova fino a quando le spine di acetato di cellulosa sono 2 cm sotto le cime delle fiale. </li><li> Hereafter, gestire ogni fila di quattro fiale come insieme ad sfalsati min intervalli di 1. </li><li> Per la valutazione tempo 0, cogliere ogni flaconcino individualmente with pollice e l&#39;indice, toccare delicatamente sul tavolo tre volte per battere mosche al fondo della fiala, attendere 30 sec e poi contare il numero di mosche che sono immobili / morti. Registrare il numero di mosche immobili / morte per ogni flacone al tempo 0 min nel registro test cartaceo. </li><li> Inizia timer conteggio continuamente al tempo 0 e iniziare immediatamente aggiungendo 1 ml di etanolo a tappi acetato di cellulosa in fiale per la prima riga / set di 4 fiale. Aggiungere 1 ml di etanolo per i tappi di acetato di cellulosa in fiale a intervalli di 5 sec nell&#39;ordine in cui saranno testati. Aggiungere etanolo per i tappi di acetato di cellulosa in un movimento circolare in modo che l&#39;etanolo viene assorbita in modo uniforme in tutto i tappi di acetato di cellulosa. Quando l&#39;etanolo è stato aggiunto a tutti i 4 flaconcini di test nel set, inserire un tappo di silicone in ogni fiala per sigillarlo. </li><li> A volte 1, 2, 3, 4 e 5 min, aggiungere 1 ml di etanolo al secondo, terzo, quarto, quinto e sesto set di 4 flaconcini, rispettivamente. Continuare inserendo tappi in silicone dopoaggiungendo etanolo per ciascun set di 4 fiale. </li><li> Al momento 6 min, testare la prima serie di 4 flaconi afferrando ogni flacone con il pollice e l&#39;indice, picchiettando delicatamente sul tavolo tre volte per battere mosche al fondo del flacone, in attesa 30 sec e poi contare e registrare il numero totale di mosche che sono sedati. Score vola come sedato se (i) stanno sul pavimento del flacone, ma non camminano o (ii) si trovano sulla schiena, con o senza sbattere le ali. </li><li> Maneggiare ogni flacone all&#39;interno del set ad intervalli di 5 sec utilizzando il programma in tabella 2. </li><li> A volte 7, 8, 9, 10 e 11 min, testare il secondo, terzo, quarto, quinto e sesto set di fiale, rispettivamente, come fatto per la prima serie. </li><li> Al tempo 12 min, testare la prima serie di 4 provette nuovamente come descritto in 2.12 e continuare il test secondo, terzo, quarto, quinto e sesto serie di flaconi a 13, 14, 15, 16 e 17 minuti, rispettivamente. </li><li> Continuare mosche test come descritto in 2.12 fino a quando tutte le mosche sono sedated. </li><li> Inserire il numero totale di mosche in ogni fiala nel registro test cartaceo. Censore mosche immobile / morti al tempo 0 dal numero totale di mosche. </li><li> Calcolare i cento mosche attivi ad ogni valutazione di time-point e dati plot come% mosche attivi (asse y) in funzione del tempo (asse x). Quantificare etanolo sedazione interpolando sedazione Tempo 50 valori (ST50, tempo al 50% sedazione) dal terzo ordine curva polinomiale o sigmoidale adatta o calcolare l&#39;area sotto la curva. </li><li> Compilare i dati provenienti da flaconi decodificati e effettuare analisi statistiche (ad esempio, una via ANOVA con Bonferroni test di confronto multiplo), secondo il disegno sperimentale. </li></ol>

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The authors have nothing to disclose.

Test semplice che riproducibile quantificare fenotipi significativi sono di grande valore per l&#39;analisi del comportamento. Il lavoro qui descritto affronta diversi aspetti pratici di un test per misurare la sensibilità etanolo sedazione e tolleranza rapida in Drosophila. Sebbene non sia un obiettivo di questo lavoro, analisi comportamentali sono facilitate mantenendo l&#39;ambiente e lo sfondo costante genetico per soggetti di prova all&#39;interno di uno studio. Inoltre, i confronti dovrebbero generalment...

L&#39;uso di alcol disturbo (AUD) è un problema di salute enorme in tutto il mondo (rivisto in 1). Sebbene i meccanismi di guida lo sviluppo di AUD sono complesse, questi disturbi hanno una notevole componente genetica (ad esempio, 2). La grande ereditabilità di AUD e le risposte comportamentali conservati in etanolo in molte specie (recensito in 3,4) hanno generato un forte interesse ad utilizzare organismi modello genetici per studiare il coinvolgimento di specifici geni nei comportamenti etanolo legate verso una migliore comprensione delle basi molecolari di AUD. Il moscerino della frutta, Drosophila melanogaster, è emerso come organismo modello principale per esplorare i meccanismi molecolari-genetica dei comportamenti etanolo correlati (recensito in 3,4). Studi in mosche hanno evidenziato i ruoli per diverse vie di segnalazione nelle risposte comportamentali a etanolo (recensito in 5). Curiosamente, alcuni dei geni e dei percorsi che influenzare respons comportamentalies a etanolo in linea sono stati implicati in roditori comportamenti etanolo correlati e / o AUD umana (ad esempio, 6-14). La conservazione di meccanismi di guida comportamenti etanolo legate tra le specie, insieme con la suite di strumenti genetici disponibili nel sistema modello Drosophila, sottolineano l&#39;utilità del modello di mosca della frutta per studiare la genetica di risposte comportamentali a etanolo. Sensibilità 15,16 e tolleranza (rivisto in 17) di etanolo negli esseri umani è legato allo sviluppo di AUD. Entrambe queste risposte comportamentali a etanolo può essere modellato in linea attraverso una serie di test di laboratorio (rivisto in 3,4). Tutti i saggi mosca noti gli autori sono basati su uno dipendente dal tempo etanolo-indotta sedazione / incoordinazione o di recupero in funzione del tempo da etanolo sedazione. In un precedente articolo dal nostro gruppo sulla genetica della sensibilità etanolo e rtolleranza APID in Drosophila, un test comportamentale basato su etanolo sedazione vapore indotta di mosche è stato utilizzato 18. Test in questo test è stato avviato con il trasferimento dal vivo adulti vola senza anestesia svuotare fiale alimentari, intrappolando le mosche nelle fiale con una spina di acetato di cellulosa, aggiungendo etanolo verso l&#39;alto (cioè, lato non volo) del tappo di acetato di cellulosa, e sigillare la fiala contenente mosche, spina acetato di cellulosa e di etanolo con un tappo di silicone (vedi schema in Figura S3, riferimento 18). Più fiale che rappresentano diversi gruppi di mosche sono state valutate in parallelo, aumentando la velocità di questo test. Fiale sono stati dati un codice anonimo e sperimentatori sono stati accecati per gruppo di trattamento per prevenire la distorsione intenzionale nella valutazione della sedazione. In un esperimento standard mosche in fiale erano picchiettare leggermente ad intervalli di 6 min e, dopo un recupero di 30 secondi, il numero di mosche sedati in ogni flacone è stato contato e converted per cento mosche attivi. Mosche assorbiti vapori di etanolo dal tappo di acetato di cellulosa in modo dipendente dal tempo, causando un aumento progressivo in etanolo interno 18 e sedazione (riferimento cf 18 e Figure 1A e 1B in questo rapporto). La sedazione in questo test è stato operativamente definita come mosche (i) in piedi in assenza di camminare o (ii) che giace sulla schiena, con o senza sbattere le ali. Qui, questo dosaggio etanolo sedazione è descritta in dettaglio, ulteriore ottimizzazione operativa che riguardano l&#39;uso è previsto, e il test viene utilizzato per indirizzare il contributo delle opzioni integrazione alimentare sulla sensibilità fly sedazione.

<table><tbody><tr><td>food vials</td><td>VWR</td><td>89092-772</td><td>narrow</td></tr><tr><td>Flugs</td><td>Genesee/flystuff.com</td><td>49-102</td><td>narrow</td></tr><tr><td>silicone stopper</td><td>Fisher Scientific</td><td>09-704-1l</td><td>#4</td></tr><tr><td>ethanol</td><td>Pharmaco-Aaper</td><td>111000200</td><td>200 proof</td></tr></tbody></table>

an inexpensive, scalable behavioral assay for measuring ethanol sedation sensitivity and rapid tolerance in drosophila

L&#39;uso di alcol disturbo (AUD) è una sfida di salute gravi. Nonostante una grande componente ereditaria per AUD, pochi geni sono stati inequivocabilmente implicati nella loro eziologia. Il moscerino della frutta, Drosophila melanogaster, è un modello potente per esplorare i meccanismi molecolari alla base genetica comportamenti alcol-correlati e detiene quindi una grande promessa per l&#39;identificazione e la comprensione della funzione dei geni che influenzano AUD. L&#39;uso del modello Drosophila per questi tipi di studi dipende dalla disponibilità di saggi che misurano affidabile risposte comportamentali a etanolo. Questo rapporto descrive un test adatto per valutare la sensibilità etanolo e tolleranza rapida in mosche. Etanolo sensibilità misurata in questo saggio è influenzata dal volume e la concentrazione di etanolo utilizzato, una varietà di manipolazioni genetiche precedentemente riportati, e anche la lunghezza di tempo in linea sono alloggiati senza cibo immediatamente prima del test. Al contrario, etanolo sensitivity misurata in questo dosaggio non è influenzato dal vigore di movimentazione mosca, sesso in linea, e la supplementazione di mezzo di crescita con antibiotici o lieviti vivi. Tre metodi diversi per quantificazione sensibilità etanolo sono descritte, tutto conduce a risultati sostanzialmente indistinguibili sensibilità etanolo. La natura scalabile di questo test, in combinazione con la sua semplicità complessiva di set-up e relativamente a basso costo, lo rendono adatto per l&#39;analisi piccola e grande scala genetica della sensibilità etanolo e tolleranza rapida in Drosophila.

I dati grezzi di questa analisi etanolo sedazione sono i numeri di mosche che vengono sedati in funzione del tempo di esposizione al vapore etanolo. I dati grezzi sono convertiti in linea attivi cento in funzione del tempo (dati primario, figure 1A, B, D - F). La sensibilità all&#39;etanolo sedazione dai dati primari può essere quantificata come sedazione Tempo 50 (ST50), il tempo necessario per il 50% di mosche per diventare sedato o aea sotto la curva (AUC), via interpolazione curva...

Watch this Scientific Journal Video about An Inexpensive, Scalable Behavioral Assay for Measuring Ethanol Sedation Sensitivity and Rapid Tolerance in Drosophila at JoVE.com

An Inexpensive, Scalable Behavioral Assay for Measuring Ethanol Sedation Sensitivity and Rapid Tolerance in Drosophila

Straightforward assays for measuring ethanol sensitivity and rapid tolerance in Drosophila facilitate the use of this model organism for investigating these important ethanol-related behaviors. Here, a relatively simple, scalable assay for measuring ethanol sensitivity and rapid tolerance in flies is described.

Un poco costoso, test comportamentale scalabile per la misurazione etanolo sedazione sensibilità e tolleranza Rapid in<em&gt; Drosophila</em

Alcohol use disorder (AUD) is a serious health challenge. Despite a large hereditary component to AUD, few genes have been unambiguously implicated in ...

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Universidad de Cartagena UDEC

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Neuroscience

An Inexpensive, Scalable Behavioral Assay for Measuring Ethanol Sedation Sensitivity and Rapid Tolerance in Drosophila

10.2K Views.  Virginia Commonwealth University. Straightforward assays for measuring ethanol sensitivity and rapid tolerance in Drosophila facilitate the use of this model organism for investigating these important ethanol-related behaviors. Here, a relatively simple, scalable assay for measuring ethanol sensitivity and rapid tolerance in flies is described.

Video: An Inexpensive, Scalable Behavioral Assay for Measuring Ethanol Sedation Sensitivity and Rapid Tolerance in Drosophila

Combining Behavioral Endocrinology and Experimental Economics: Testosterone and Social Decision Making

Behavioral endocrinological research in humans as well as in animals suggests that testosterone plays a key role in social interactions. Studies in rodents have shown a direct link between testosterone and aggressive behavior1 and folk wisdom adapts these findings to humans, suggesting that testosterone induces antisocial, egoistic or even aggressive behavior2. However, many researchers doubt a direct testosterone-aggression link in humans, arguing instead that testosterone is primarily involved in status-related behavior3,4. As a high status can also be achieved by aggressive and antisocial means it can be difficult to distinguish between anti-social and status seeking behavior.
We therefore set up an experimental environment, in which status can only be achieved by prosocial means. In a double-blind and placebo-controlled experiment, we administered a single sublingual dose of 0.5 mg of testosterone (with a hydroxypropyl-&beta;-cyclodextrin carrier) to 121 women and investigated their social interaction behavior in an economic bargaining paradigm. Real monetary incentives are at stake in this paradigm; every player A receives a certain amount of money and has to make an offer to another player B on how to share the money. If B accepts, she gets what was offered and player A keeps the rest. If B refuses the offer, nobody gets anything. A status seeking player A is expected to avoid being rejected by behaving in a prosocial way, i.e. by making higher offers.
The results show that if expectations about the hormone are controlled for, testosterone administration leads to a significant increase in fair bargaining offers compared to placebo. The role of expectations is reflected in the fact that subjects who report that they believe to have received testosterone make lower offers than those who say they believe that they were treated with a placebo. These findings suggest that the experimental economics approach is sensitive for detecting neurobiological effects as subtle as those achieved by administration of hormones. Moreover, the findings point towards the importance of both psychosocial as well as neuroendocrine factors in determining the influence of testosterone on human social behavior.

The procedure described in this protocol shows that testosterone administration and folk beliefs about testosterone may be associated with directly opposed social behaviors.

Combinando comportamentali Endocrinologia ed Economia Sperimentale: Decisione testosterone e sociale Fare

Behavioral endocrinological research in humans as well as in animals suggests that testosterone plays a key role in social interactions. Studies in ...

Ricerca

Neuroscienze

A Simple Way to Measure Ethanol Sensitivity in Flies

Low doses of ethanol cause flies to become hyperactive, while high doses are sedating. The sensitivity to ethanol-induced sedation of a given fly strain is correlated with that strain s ethanol preference, and therefore sedation is a highly relevant measure to study the genetics of alcohol responses and drinking. We demonstrate a simple way to expose flies to ethanol and measure its intoxicating effects. The assay we describe can determine acute sensitivity, as well as ethanol tolerance induced by repeat exposure. It does not require a technically involved setup, and can therefore be applied in any laboratory with basic fly culture tools.

A simple assay to measure the sedating effects of ethanol on Drosophila flies, based on the loss of righting reflex, is described.

Un modo semplice per misurare sensibilità etanolo in Flies

Low doses of ethanol cause flies to become hyperactive, while high doses are sedating. The sensitivity to ethanol-induced sedation of a given fly strain ...

A General Method for Evaluating Incubation of Sucrose Craving in Rats

For someone on a food-restricted diet, food craving in response to food-paired cues may serve as a key behavioral transition point between abstinence and relapse to food taking 1. Food craving conceptualized in this way is akin to drug craving in response to drug-paired cues. A rich literature has been developed around understanding the behavioral and neurobiological determinants of drug craving; we and others have been focusing recently on translating techniques from basic addiction research to better understand addiction-like behaviors related to food 2-4.
As done in previous studies of drug craving, we examine sucrose craving behavior by utilizing a rat model of relapse. In this model, rats self-administer either drug or food in sessions over several days. In a session, lever responding delivers the reward along with a tone+light stimulus. Craving behavior is then operationally defined as responding in a subsequent session where the reward is not available. Rats will reliably respond for the tone+light stimulus, likely due to its acquired conditioned reinforcing properties 5. This behavior is sometimes referred to as sucrose seeking or cue reactivity. In the present discussion we will use the term "sucrose craving" to subsume both of these constructs.
In the past decade, we have focused on how the length of time following reward self-administration influences reward craving. Interestingly, rats increase responding for the reward-paired cue over the course of several weeks of a period of forced-abstinence. This "incubation of craving" is observed in rats that have self-administered either food or drugs of abuse 4,6. This time-dependent increase in craving we have identified in the animal model may have great potential relevance to human drug and food addiction behaviors.
Here we present a protocol for assessing incubation of sucrose craving in rats. Variants of the procedure will be indicated where craving is assessed as responding for a discrete sucrose-paired cue following extinction of lever pressing within the sucrose self-administration context (Extinction without cues) or as responding for sucrose-paired cues in a general extinction context (Extinction with cues).

Responding for food or drug-paired cues increases over the course of a period of abstinence and this may relate to an increased susceptibility to relapse behaviors. Here we detail a procedure for evaluating this "incubation of craving" in rats that have self-administered sucrose.

Un metodo generale per la valutazione di incubazione della brama di saccarosio in ratti

For someone on a food-restricted diet, food craving in response to food-paired cues may serve as a key behavioral transition point between abstinence and ...

Recording and Analysis of Circadian Rhythms in Running-wheel Activity in Rodents

When rodents have free access to a running wheel in their home cage, voluntary use of this wheel will depend on the time of day1-5. Nocturnal rodents, including rats, hamsters, and mice, are active during the night and relatively inactive during the day. Many other behavioral and physiological measures also exhibit daily rhythms, but in rodents, running-wheel activity serves as a particularly reliable and convenient measure of the output of the master circadian clock, the suprachiasmatic nucleus (SCN) of the hypothalamus. In general, through a process called entrainment, the daily pattern of running-wheel activity will naturally align with the environmental light-dark cycle (LD cycle; e.g. 12 hr-light:12 hr-dark). However circadian rhythms are endogenously generated patterns in behavior that exhibit a ~24 hr period, and persist in constant darkness. Thus, in the absence of an LD cycle, the recording and analysis of running-wheel activity can be used to determine the subjective time-of-day. Because these rhythms are directed by the circadian clock the subjective time-of-day is referred to as the circadian time (CT). In contrast, when an LD cycle is present, the time-of-day that is determined by the environmental LD cycle is called the zeitgeber time (ZT).
Although circadian rhythms in running-wheel activity are typically linked to the SCN clock6-8, circadian oscillators in many other regions of the brain and body9-14 could also be involved in the regulation of daily activity rhythms. For instance, daily rhythms in food-anticipatory activity do not require the SCN15,16 and instead, are correlated with changes in the activity of extra-SCN oscillators17-20. Thus, running-wheel activity recordings can provide important behavioral information not only about the output of the master SCN clock, but also on the activity of extra-SCN oscillators. Below we describe the equipment and methods used to record, analyze and display circadian locomotor activity rhythms in laboratory rodents.

Circadian rhythms in voluntary wheel-running activity in mammals are tightly coupled to the molecular oscillations of a master clock in the brain. As such, these daily rhythms in behavior can be used to study the influence of genetic, pharmacological, and environmental factors on the functioning of this circadian clock.

Registrazione e analisi dei ritmi circadiani in Running-ruota Attività nei roditori

When rodents have free access to a running wheel in their home cage, voluntary use of this wheel will depend on the time of day1-5. Nocturnal rodents, ...

The FlyBar: Administering Alcohol to Flies

Fruit flies (Drosophila melanogaster) are an established model for both alcohol research and circadian biology. Recently, we showed that the circadian clock modulates alcohol sensitivity, but not the formation of tolerance. Here, we describe our protocol in detail. Alcohol is administered to the flies using the FlyBar. In this setup, saturated alcohol vapor is mixed with humidified air in set proportions, and administered to the flies in four tubes simultaneously. Flies are reared under standardized conditions in order to minimize variation between the replicates. Three-day old flies of different genotypes or treatments are used for the experiments, preferably by matching flies of two different time points (e.g., CT 5 and CT 17) making direct comparisons possible. During the experiment, flies are exposed for 1 hr&#160;to the pre-determined percentage of alcohol vapor and the number of flies that exhibit the Loss of Righting reflex (LoRR) or sedation are counted every 5 min. The data can be analyzed using three different statistical approaches. The first is to determine the time at which 50% of the flies have lost their righting reflex and use an Analysis of the Variance (ANOVA) to determine whether significant differences exist between time points. The second is to determine the percentage flies that show LoRR after a specified number of minutes, followed by an ANOVA analysis. The last method is to analyze the whole times series using multivariate statistics. The protocol can also be used for non-circadian experiments or comparisons between genotypes.

Drosophila has emerged as a significant model system for dissecting the cellular and molecular underpinnings of behavioral responses to alcohol. Here we present a protocol for the collection of alcohol sensitivity data in a circadian context that can be easily applied to other experiments and is well-suited for undergraduate research.

Flybar: Amministrazione alcol a Flies

Fruit flies (Drosophila melanogaster) are an established model for both alcohol research and circadian biology. Recently, we showed that the circadian ...

High-Throughput Method for Measuring Alcohol Sedation Time of Individual Drosophila melanogaster

Drosophila melanogaster provides an excellent model to study the genetic underpinnings of alcohol sensitivity. In contrast to studies in human populations, the Drosophila model allows strict control over genetic background, and virtually unlimited numbers of individuals of the same genotype can be reared rapidly under well-controlled environmental conditions without regulatory restrictions and at relatively low cost. Flies exposed to ethanol undergo physiological and behavioral changes that resemble human alcohol intoxication, including loss of postural control, sedation, and development of tolerance. Here, we describe a simple, low-cost, high-throughput assay for assessing alcohol sedation sensitivity in large numbers of single flies. The assay is based on video recording of single flies introduced without anesthesia in 24-well cell culture plates in a set-up that enables synchronous initiation of alcohol exposure. The system enables a single person to collect individual ethanol sedation data on as many as 2,000 flies within an 8 h work period. The assay can, in principle, be extended to assess the effects of exposure to any volatile substance and applied to measure effects of acute toxicity of volatiles on other insects, including other fly species.

High-Throughput Method for Measuring Alcohol Sedation Time of Individual Drosophila melanogaster

Current methods to measure alcohol sensitivity in Drosophila are designed to test groups of flies. We present a simple, low-cost, high-throughput assay for assessing alcohol sedation sensitivity in large numbers of single flies. The method does not require specialized tools and can be performed in any laboratory using common materials.

Metodo ad alta velocità effettiva per la misurazione del tempo di sedazione alcolica del melanogaster di Drosophila individuale

Drosophila melanogaster provides an excellent model to study the genetic underpinnings of alcohol sensitivity. In contrast to studies in human ...

Comportamento

Flypub To Study Ethanol Induced Behavioral Disinhibition and Sensitization

Alcohol use disorder (AUD) remains a serious problem in our society. To develop effective interventions for addiction, it is important to understand the underlying neurobiological mechanisms, for which diverse experimental approaches and model systems are needed. The main ingredient of alcoholic beverages is ethanol, which causes adaptive changes in the central nervous system and behavior upon chronic intake. Behavioral sensitization (i.e., escalated responses) in particular represents a key adaptive change underlying addiction. Most ethanol-induced behavioral sensitization studies in animal models have been conducted on the locomotor activating effect of ethanol. A prominent effect of ethanol is behavioral disinhibition. Behavioral sensitization to the disinhibition effect of ethanol, however, is underrepresented. To address this issue, we developed the Flypub assay that allows measuring the escalated increase in disinhibited courtship activities upon recurring ethanol exposure in Drosophila melanogaster.&#160;Here, we report the step-by-step Flypub assay including assembly of ethanol exposure chambers, setup of the assay station, criteria for fly care and collection, ethanol delivery, quantification of disinhibited courtship activities, data processing and statistical analysis. Also provided are how to troubleshoot critical steps, overcome limitations and expand its utility to assess additional ethanol-induced behaviors. The Flypub assay in combination with powerful genetic tools in Drosophila melanogaster will facilitate the task of discovering the mechanism underlying ethanol-induced behavioral sensitization.

The Flypub assay measures the behaviors that the fruit fly Drosophila melanogaster displays under the influence of ethanol. The assay can be readily mastered by experimenters at all levels and applied to various vaporized stimuli, facilitating substance abuse and addiction studies.

Flypub Per studiare l'etanolo indotto disinibie comportamentali e sensibilitazione

Alcohol use disorder (AUD) remains a serious problem in our society. To develop effective interventions for addiction, it is important to understand the ...

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.

Assay Semplice per la quantificazione della Social Prevenzione in<em&gt; Drosophila melanogaster</em

Drosophila melanogaster is an emerging model to study different aspects of social interactions. For example, flies avoid areas previously occupied by ...

An Assay for Measuring the Effects of Ethanol on the Locomotion Speed of Caenorhabditis elegans

Alcohol use disorders are a significant public health concern, for which there are few effective treatment strategies. One difficulty that has delayed the development of more effective treatments is the relative lack of understanding of the molecular underpinnings of the effects of ethanol on behavior. The nematode, Caenorhabditis elegans (C.&#160;elegans), provides a useful model in which to generate and test hypotheses about the molecular effects of ethanol. Here, we describe an assay that has been developed and used to examine the roles of particular genes and environmental factors in behavioral responses to ethanol, in which locomotion is the behavioral output. Ethanol dose-dependently causes an acute depression of crawling on an agar surface. The effects are dynamic; animals exposed to a high concentration demonstrate an initial strong depression of crawling, referred to here as initial sensitivity, and then partially recover locomotion speed despite the continued presence of the drug. This ethanol-induced behavioral plasticity is referred to here as the development of acute functional tolerance. This assay has been used to demonstrate that these two phenotypes are distinct and genetically separable. The straightforward locomotion assay described here is suitable for examining the effects of both genetic and environmental manipulations on these acute behavioral responses to ethanol in C.&#160;elegans.

An Assay for Measuring the Effects of Ethanol on the Locomotion Speed of Caenorhabditis elegans

C. elegans is a useful model for studying the effects of ethanol on behavior. We present a behavioral assay that quantifies the effects of ethanol on the locomotion speed of crawling worms; both initial sensitivity and the development of acute functional tolerance to ethanol can be measured with this assay.

Un test per misurare gli effetti di etanolo sulla velocità di Locomotion<em&gt; Caenorhabditis elegans</em

Alcohol use disorders are a significant public health concern, for which there are few effective treatment strategies. One difficulty that has delayed the ...

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 ...

Un poco costoso, test comportamentale scalabile per la misurazione etanolo sedazione sensibilità e tolleranza Rapid in

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