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. Day Before Assay <ol><li> Recolha voa em frascos de alimentos frescos em grupos de 11 (do mesmo sexo), sob breve (1-5 min) CO 2. </li><li> Permitir moscas para recuperar S / N em frascos de alimentos num espaço de ambiente controlado (tipicamente 25 ° C, 60% de humidade relativa, 12 horas de luz / escuridão). </li><li> Preparar a solução de (s) diluindo etanol puro (100%) em etanol purificado (≥18 mohms) de água para a concentração (s) final adequada para a experiência planeada. Permitir que a solução (s) para voltar à TA O / N. Nota: Diluição de etanol é exotérmica. </li></ol> 2. Dia de Ensaio <ol><li> Para cada frasco de moscas para ser testado, preparar (i) um frasco vazio alimentos limpo; ou seja, testes de frasco, (ii) uma nova ficha de acetato de celulose, (iii) uma rolha de silicone e (iv) 1 ml de solução de etanol (ver Tabela 3). </li><li> Prepare a sala de ensaio, ajustando a temperatura de 20-25 ° C e humidade relativa de 55-65%. </li><li> Já um outro trabalhadoratribuir um código único para cada grupo de frascos e registrar o código para mais tarde. Coloque frascos codificados com moscas na sala de testes para se aclimatar para poucos min. </li><li> Rotular frascos vazios de teste para coincidir com códigos em frascos de voar a partir de 2.3. </li><li> Construir um log de ​​teste impressa, digitando os códigos em colunas (uma coluna / frasco) em uma planilha semelhante à Tabela 1. </li><li> Usando o log de teste como um guia, organizar frascos de alimentos codificados com moscas e frascos vazios de testes em matrizes correspondentes na sala de testes. Nota: Um número máximo razoável de frascos de testar é 24 (ou seja, 6 conjuntos de 4 frascos cada). </li><li> Transferência voa de frascos de alimentos em pareados / rotulados frascos de ensaio vazio e coloque imediatamente plugs de acetato de celulose em frascos de ensaio até plugs de acetato de celulose são 2 cm abaixo dos topos dos frascos. </li><li> A partir de agora, lidar com cada linha de quatro frascos como um conjunto em escalonados intervalos de 1 min. </li><li> Para a avaliação do tempo 0, segure cada frasco individualmente wom polegar eo indicador, bater suavemente sobre a mesa três vezes para bater as moscas para o fundo do frasco, esperar 30 segundos e depois contar o número de moscas que são imóveis / morto. Grave o número de moscas mortas / imóveis para cada frasco no tempo 0 min no log de teste impresso. </li><li> Comece contador de tempo de forma contínua no tempo 0 e começar imediatamente a adição de 1 ml de etanol para plugs de acetato de celulose nos frascos para a primeira linha / set de 4 frascos. Adicionar 1 ml de etanol para as velas de acetato de celulose nos frascos a intervalos de 5 seg na ordem em que irá ser testada. Adicionar etanol para os tampões de acetato de celulose em um movimento circular de modo que o etanol é absorvido uniformemente ao longo dos tampões de acetato de celulose. Quando o etanol foi adicionado a todos os 4 frascos de ensaio no conjunto, insira um plug silicone em cada frasco para selá-lo. </li><li> Às vezes 1, 2, 3, 4 e 5 min, adicionar 1 ml de etanol para o segundo, terceiro, quarto, conjuntos, quinto e sexto de 4 frascos, respectivamente. Continue inserir rolhas de silicone apósadição de etanol para cada conjunto de 4 frascos. </li><li> Em vez de 6 min, testar o primeiro conjunto de quatro frascos de agarrar cada frasco com o polegar eo dedo indicador, batendo suavemente na mesa três vezes para bater as moscas para o fundo do frasco, de espera de 30 segundos e, em seguida, a contagem e registo do número total de moscas que estão sedados. Score voa tão sedado se (i) estar no chão do frasco, mas não andam ou (ii) se situam em suas costas com ou sem bater as asas. </li><li> Lidar com cada frasco dentro do conjunto em intervalos de 5 seg usando o cronograma na Tabela 2. </li><li> Às vezes, 7, 8, 9, 10 e 11 min, testar as segunda, terceira, quarta, quinta e sexta séries de frascos, respectivamente, como é feito com o primeiro set. </li><li> Em tempo de 12 min, testar o primeiro conjunto de 4 frascos novamente como descrito em 2.12 e continuar testando o segundo terceiro, quarto sets, quinto e sexto de frascos em 13, 14, 15, 16 e 17 min, respectivamente. </li><li> Continuar ensaio moscas como descrito em 2,12 até que todas as moscas são sedated. </li><li> Digite o número total de moscas em cada frasco no log de testes cópia impressa. Censor moscas imóveis / mortas no tempo 0 a partir do número total de moscas. </li><li> Calcular a percentagem de moscas ativas em cada tempo de ponto de avaliação e dados de enredo como% moscas ativas (eixo y) em função do tempo (eixo-x). Quantificar sedação etanol interpolando Sedação Tempo 50 valores (ST50, tempo para 50% sedação) de terceira ordem curva polinomial ou sigmoidal encaixa ou área de cálculo sob a curva. </li><li> Compilar dados de frascos decodificados e realizar análises estatísticas (por exemplo, one-way ANOVA com Bonferroni teste de comparação múltipla) conforme apropriado para o projeto experimental. </li></ol>

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The authors have nothing to disclose.

Ensaios simples que reproducibly quantificada fenótipos significativas são de grande valor para a análise do comportamento. O trabalho descrito aqui aborda vários aspectos práticos de um ensaio para medir a sensibilidade sedação etanol e tolerância rápida em Drosophila. Apesar de não ser um foco deste trabalho, análises de comportamento são facilitadas pela manutenção do ambiente e genético constante fundo para indivíduos de teste dentro de um estudo. Além disso, comparações devem tipicamente...

Transtorno de uso de álcool (AUD) é um enorme problema de saúde mundial (revisto em 1). Embora os mecanismos que conduzem o desenvolvimento de R são complexas, estas perturbações têm um importante componente genético (por exemplo, 2). O grande herdabilidade de AUD e as respostas comportamentais conservados em etanol através de muitas espécies (revisto em 3,4) têm gerado grande interesse na utilização de organismos-modelo genéticos para investigar o envolvimento de genes específicos em comportamentos relacionados com o álcool para uma melhor compreensão das bases moleculares de AUD. A mosca da fruta, Drosophila melanogaster, surgiu como um organismo modelo de liderança para explorar mecanismos genético-moleculares de comportamentos relacionados com o etanol (revisto em 3,4). Estudos em moscas têm destacado papel para várias vias de sinalização em respostas comportamentais ao etanol (revisto em 5). Curiosamente, alguns dos genes e vias que influenciam respons comportamentaises para etanol em moscas também têm sido implicados em comportamentos relacionados com etanol roedores e / ou R humano (por exemplo, 6-14). A conservação dos mecanismos de condução comportamentos relacionados com o etanol em toda a espécie, juntamente com o conjunto de ferramentas genéticas disponíveis no sistema modelo de Drosophila, ressaltam a utilidade do modelo mosca da fruta para investigar a genética de respostas comportamentais ao etanol. Sensibilidade e tolerância 15,16 (revisto em 17) ao etanol em seres humanos está associada com o desenvolvimento de AUD. Ambas as respostas comportamentais ao etanol pode ser modelado em moscas através de uma variedade de ensaios laboratoriais (revisto em 3,4). Todos os ensaios de mosca conhecidos para os autores são baseados quer dependente do tempo induzida pelo etanol sedação / ou descoordenação recuperação dependente do tempo a partir de etanol sedação. Em um artigo anterior do nosso grupo sobre a genética da sensibilidade etanol e rtolerância APID em Drosophila, um ensaio comportamental, baseada em etanol sedação induzida por vapor de moscas foi utilizado 18. Testes neste ensaio foi iniciado através da transferência de adulto vivo voa sem anestesia para esvaziar frascos alimentares, aprisionando as moscas em frascos com um tampão de acetato de celulose, adição de etanol para a parte superior (ou seja, o lado não-fly) do tampão de acetato de celulose, e selando o frasco contendo moscas, plugue de acetato de celulose e de etanol com uma rolha de silicone (ver esquema na figura S3, Referência 18). Vários frascos que representam diferentes grupos de moscas foram avaliados em paralelo, aumenta o rendimento do presente ensaio. Os frascos foram dadas um código anônimo e experimentadores ficaram cegos para grupo de tratamento para evitar viés não intencional na avaliação da sedação. Numa experiência normal, moscas em frascos foram bate-se suavemente a intervalos de 6 min e, após uma recuperação de 30 seg, o número de moscas sedados em cada frasco foi contado e Converted para moscas por cento ativos. Moscas absorvida vapor de etanol a partir do tampão de acetato de celulose de uma maneira dependente do tempo, causando aumentos progressivos em etanol interna 18 e sedação (cf referência 18 e Figura 1A e 1B neste relatório). Sedação neste ensaio foi definida operacionalmente como moscas (i) que está na ausência de curta ou (ii) que encontra-se em suas costas com ou sem bater as asas. Aqui, este ensaio de sedação etanol é descrita em detalhe, uma maior optimização operacional relevante para usá-lo é fornecido, e o ensaio é utilizado para tratar a contribuição de opções de suplementação de alimentos sobre a sensibilidade mosca sedação.

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

Transtorno de uso de álcool (AUD) é um desafio de saúde grave. Apesar de uma grande componente hereditário para AUD, alguns genes têm sido implicados de forma inequívoca na sua etiologia. A mosca da fruta, Drosophila melanogaster, é um modelo poderoso para explorar mecanismos genético-moleculares subjacentes comportamentos relacionados com o álcool e, portanto, é uma grande promessa para identificar e compreender a função dos genes que influenciam AUD. A utilização do modelo de Drosophila para estes tipos de estudos depende da disponibilidade de ensaios que medem a forma fiável respostas comportamentais ao etanol. Este relatório descreve um ensaio adequado para avaliar a sensibilidade de etanol e tolerância rápida em moscas. Sensibilidade etanol medido neste ensaio é influenciado pelo volume e concentração do etanol usado, uma variedade de manipulações genéticas previamente relatados, e também o período de tempo em que as moscas são alojados sem alimentos imediatamente antes do teste. Em contraste, s etanolensitivity medido neste ensaio não é afectado pela força de manipulação mosca, sexo das moscas, e a suplementação do meio de crescimento com antibióticos ou com levedura viva. Três métodos diferentes para quantificar a sensibilidade de etanol encontram-se descritos, todos levando a resultados essencialmente indistinguíveis sensibilidade etanol. A natureza escalável deste ensaio, combinada com a sua simplicidade global de set-up e custo relativamente baixo, tornam adequado para a análise genética de pequena escala e grande sensibilidade de etanol e tolerância rápida em Drosophila.

Os dados em bruto a partir de etanol neste ensaio de sedação são os números de moscas que são sedados como uma função do tempo de exposição ao vapor de etanol. Os dados em bruto são convertidos para as moscas por cento ativos como uma função do tempo (dados primários, as Figuras 1A, B, D - F). Sensibilidade para sedação de etanol a partir dos dados primários podem ser quantificados como sedação 50 Tempo (ST50), o tempo necessário para que 50% das moscas de se tornar s...

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.

Um Ensaio Behavioral Barato, escalável para medição Ethanol Sedação sensibilidade e tolerância rápida em<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 ...

an-inexpensive-scalable-behavioral-assay-for-measuring-ethanol

Research

JoVE Journal

Neuroscience

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

10.2K visualizações.  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.

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

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.

Uma maneira simples para medir a sensibilidade de etanol no 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 ...

Neurociência

Methods to Assay Drosophila Behavior

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

Methods to Assay Drosophila Behavior

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

Métodos de Ensaio para Drosophila Comportamento

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

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.

O Flybar: Administração álcool para Moscas

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

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 Ensaio para comportamento de forrageamento em<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 ...

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.

Método de alta duração para medir o tempo de sedação de álcool de melanogaster individual drosophila

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 estuda desinibição comportamental induzida pelo etanol e sensibilização

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.

Ensaio simples para Quantificação de Prevenção Social<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.

Um ensaio para medir os efeitos do etanol sobre a velocidade de 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.

Uma maneira simples para medir alterações na Recompensa comportamento de busca Usando<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 ...

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.

Um ensaio rápido de preferência alimentar em Drosophila

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

Um Ensaio Behavioral Barato, escalável para medição Ethanol Sedação sensibilidade e tolerância rápida em

Resumo

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