Name: straightforward assay for quantification of social avoidance in drosophila melanogaster
Uploaded: 2014-12-13T14:00:00.000+00:00
Duration: 8 min 8 s
Description: Watch this Scientific Journal Video about Straightforward Assay for Quantification of Social Avoidance in Drosophila melanogaster at JoVE.com

We thank Rachelle Kanippayoor for her help in identifying the new wild-type strain as being of the melanogaster species. R.W.F, O.F. and A.F.S were responsible for research design; R.W.F, M.N. and O.F. performed the experiments. R.W.F, M.N., O.F. and A.F.S. analyzed the data; R.W.F., I.S.M. and A.F.S. wrote the manuscript. 
This work was supported by PSC-CUNY research awards, jointly funded by The Professional Staff Congress and The City University of New York to A.F.S.; by internal funding from Western University to A.F.S. and I.S.M.; by a training support from the National Alliance for Hispanic Health&#8217;s Alliance/Merck Ciencia (Science) Hispanic Scholars Program and a University Fellowship from the Yale Graduate School of Arts and Sciences to R.W.F.

1. equipamentos e reagentes criados em casa (Veja a lista de material para os outros) <ol><li> Prepare um aparelho de anestesia Drosophila frio para realizar o trabalho real. <ol><li> Cortar uma folha de polietileno poroso para cobrir uma pequena caixa de plástico (12.7 cm de comprimento, 10,2 cm de largura), normalmente a tampa superior de uma caixa de pontas de pipeta (12.7 cm de comprimento, 10,2 cm de largura, 3,8 cm de profundidade). </li><li> Preencha a caixa com gelo picado, cubra com a folha de polietileno poroso. </li></ol></li><li> Prepare um aparelho T-maze; adaptar o aparelho anteriormente descrito em detalhes 28-31. </li><li> Preparar um aspirador de mosca como descrito 32. </li><li> Prepare um aparelho de resposta phototaxis para deslocar moscas dentro e fora de frascos de teste sem estresse: <ol><li> Usar um aparelho em contracorrente, tal como descrito por 29,33. </li></ol></li></ol> NOTA: O labirinto em T pode também ser utilizado para outros fins, como descrito por 28. <ol start="5"><li> Certifique-homcondições de iluminação ogeneous: realizar o experimento em um banco coberto com uma capa de banco branco, e na frente de um quadro branco. NOTA: Para estas experiências, as linhagens de moscas utilizados foram Drosophila melanogaster tipos selvagens: Canton-S, Oregon e Samarkand voa de nossos estoques de laboratório 34; Elwood moscas foram coletadas no outono de 2011 no bairro de Elwood de Huntington, em Long Island, Nova York, EUA. </li></ol> 2. Preparar as moscas antes do experimento 1-2 dias antes de realizar o experimento: <ol><li> Manter moscas em meio padrão fubá-agar-melaço de levedura a 25 ° C em um ciclo claro / escuro de 12 horas de luz. </li><li> Recolher as moscas que respondem sob anestesia frio 1-2 dias antes de se efectuar a experiência <ol><li> Transferem-se as moscas das garrafas no qual foram colocadas em tubos Falcon de 50 ml, utilizando um funil. </li><li> Coloque o tubo de falcão com as moscas sob nível de gelo, em umbalde de gelo isolado. </li><li> Coloque o aparelho de anestesia fria com gelo picado a -4 ° C para relaxar. </li><li> Espere 5 minutos para as moscas para entrar em um coma frio. </li><li> Transferem-se as moscas para a folha de polietileno poroso do aparelho de anestesia frio. </li><li> Colete as moscas de resposta sobre o aparelho de anestesia frio. No RT, sob microscópio estereoscópico, separe masculina 3-7 dias de idade de moscas fêmeas e manter essas moscas em grupos de 40. </li></ol></li><li> Recolher as moscas emissor 1-2 dias antes de realizar a experiência. <ol><li> Gentilmente recolher Canton-S mixed-sexo voa de suas garrafas, usando um aspirador de boca, controlando o seu número. </li><li> Manter essas moscas no grupo de 60-100, pois eles serão os emissores. </li><li> Preparar todas as amostras de emissores como pontos de dados necessários. </li></ol></li><li> Permitir que pelo menos uma noite de recuperação após a coleta dos respondentes e emissores, para minimizar qualquer efeito de confusão da anestesia frioe coleta de boca aspirador no comportamento. </li><li> Abrir o saco de frascos de teste, para permitir que o ar com cheiro plástico obsoleto em que o saco seja substituído por ar fresco, pelo menos 1 dia antes da experiência. </li></ol> 2 horas antes da realização do ensaio: <ol start="6"><li> Certifique-se de que a temperatura na sala em que a experiência é realizada é de cerca de 23-25 ​​° C, com luz uniforme, e humidade superior a 30%. </li><li> Transfira os respondedores e moscas emissores de frutas em frascos de alimentos frescos para garantir que estas moscas não estão morrendo de fome. </li><li> Deixe que as moscas de fruta ajustar ao ambiente, durante 2 horas, no banco em que o experimento será realizado. </li></ol> 3. a realização do experimento evitação social <ol><li> Executar cada experiência à mesma hora do dia, num intervalo de 3-4 horas, à tarde entre o tempo Zeitgeber ZT5 e ZT9. </li><li> Coloque o T-maze no teclado batendo, e aperte o parafuso de modo clipe tele é estável elevador. </li><li> Transfira as moscas de resposta em um tubo de ensaio fresco. Encaixe este frasco contendo as moscas de resposta sobre a parte superior do labirinto-T. </li><li> Inclinar o aparelho T-maze e coloque-a sobre o tapete batendo tão moscas da fruta cair no elevador. </li><li> Mover a parte superior do elevador para baixo para que as moscas são entre a secção superior e inferior do labirinto em T, mas prestar atenção para manter o elevador acima do ponto de escolha - para impedir que as moscas de escapar. </li><li> Obter frascos com DSO (ver seção 5 para gerar DSO-free frascos que foram anteriormente ocupadas por moscas). <ol><li> Enquanto as moscas de resposta estão se ajustando a esse novo ambiente, coloque as moscas de emissor em um tubo de ensaio. </li><li> Coloque um pedaço de algodão para fechar o frasco de modo que as moscas de fruta não escapam. </li><li> Mecanicamente isto agitar o frasco numa mini-vórtice do seguinte modo: em vórtice durante 15 seg, remover o frasco de vórtice durante 5 segundos. Repita 3 vezes para um total de 55 seg. </li><li> Remove as moscas emissor para fora do frasco fresco - por transferência num frasco de alimentos - e rapidamente colocar esse frasco cheio DSO em um dos dois lados do aparelho de labirinto-T. </li><li> Alterne o lado de colocação para o frasco DSO preenchidos em cada nova execução. </li></ol></li><li> Coloque um tubo de ensaio fresco no outro lado do labirinto-T. </li><li> Traga o elevador completamente para baixo, de modo que as moscas de fruta pode escolher entre o tubo de ensaio fresco eo tubo de ensaio com DSO, e iniciar o timer. </li><li> Deixe as moscas escolher entre o frasco fresco e o frasco DSO, por 1 min, salvo indicação em contrário. </li><li> Após 1 min, mover o elevador para cima para separar as moscas no frasco fresco, DSO frasco, e os que estão presos no elevador. </li><li> Contar o número de moscas em cada frasco, e no elevador. </li><li> Repita esse procedimento para cada genótipo / condição. </li></ol> 4. Analisando os dados evitação social <ol><li> Contar o número de moscas da fruta em cada frasco para cada Genotipo, e transferir os dados para uma planilha. </li><li> Usando um programa de planilha, calcular o Índice de Desempenho (PI) para cada genótipo, subtraindo o número de moscas no frasco DSO do número de moscas no frasco DSO-livre, e dividindo pelo número total de moscas. </li><li> Compare dos IPs que consiste em utilizar o software de análise estatística para análise dos resultados. <ol><li> Idealmente, preparar 2-3 repetições internas de cada experiência, e três repetições independentes, realizadas em dias diferentes, com diferentes garrafas ou 30 atravessa. </li><li> Use testes estatísticos de distribuição de Gauss, como os dados seguem uma distribuição normal. </li><li> Ao testar um efeito (isto é, densidade - Figura 2), utilizar um ANOVA de uma via, com o pós-teste de comparação múltipla (coluna por coluna), ou um t-teste simples para comparar cada resultado de um controlo. </li><li> Quando experimentos do grupo são realizadas, testar ambas as condições e sexos (frascos vazios, destacou umd moscas un-esforçado; e efeito de gênero - Figura 1), usando uma ANOVA de duas vias. </li></ol></li></ol> 5. Gerar frascos anteriormente ocupado por Flies átonas - Realizar Em vez de 3,5-3,6 <ol><li> Use fototaxia para preparar frascos ocupados por moscas que não têm sido destacadas, como um controlo. </li><li> Transfira as moscas em um tubo de ensaio, e encaixe-o aparelho em contracorrente, com frescos frascos vazios no local oposto. </li><li> Coloque o aparelho na horizontal, com o frasco distal diretamente em frente a 15 W fluorescente luz cool-branco, e cubra com um pano preto. </li><li> Deixe as moscas mover suavemente no frasco fresco. Isso pode levar vários minutos. Uma vez que a maioria das moscas estão caminhando em direção à luz no frasco distal, iniciar o temporizador para 1 min. </li><li> Remova cuidadosamente o pano preto, mudar a localização da luz, recuperar com o pano preto, e permitir que as moscas de deixar o tubo de ensaio. </li><li> Naquela época, eunitiate o experimento escolha como em 3.2 a 3.5. Ir 3.6 e 3.7. </li><li> Mover o cursor do aparelho, para separar as moscas que desocupado o tubo de ensaio a partir do outro lado. </li><li> Use o frasco teste desocupado na T-labirinto de imediato para o experimento escolha, como em 3.7. </li></ol>

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The authors declare that they have no competing financial interests.

Este protocolo descreve um procedimento detalhado para o ensaio de evitação social. Canton-S só vai evitar um frasco em que voa tenham sido previamente stress mecânico, e que o sexo e número de respondentes que não afeta o desempenho evitação social. No entanto, a base genética dos respondentes tem uma grande influência. Seguem-se vários passos críticos para a realização deste experimento com sucesso: 1) sempre transferir as moscas 2 horas antes do experimento e certifique-se d...

O objetivo deste método é quantificar facilmente um novo aspecto do comportamento social simples em Drosophila melanogaster, independente do namoro e agressão. As interações sociais são cruciais para o bom desenvolvimento e saúde dos indivíduos dentro de uma sociedade, bem como a funcionalidade de um grupo social como um todo. A elevada complexidade destas interacções exige grandes tamanhos de amostra e um sistema que permite a simplificação do comportamento como as bases genéticas e neurais do comportamento social são ainda pouco compreendidos. Drosophila melanogaster é um poderoso modelo genético que pode ser usada para identificar os factores genéticos e bases neurais de interações sociais. Com efeito, D. melanogaster tem um repertório de comportamentos sociais complexos e algumas medições diretas de socialização já foram feitas 1-7. No entanto, a maioria desses esforços têm sido focados em comportamentos sociais relativamente complexas, como aggresinterações sive 3,6, vários aspectos do namoro 3,8-12, e como experiência social afeta outros comportamentos como a aprendizagem, ou ritmo circadiano 13-17. Além disso, muitos desses ensaios dependem de analisar os padrões de interacção complexas de grupos de moscas, usando o acompanhamento de vídeo e software de computador para analisar a abundância resultante de dados. Essas análises são de valor inestimável, e levar a novos conhecimentos importantes, como a dinâmica das interações fly-fly em grupos de sete. Uma limitação, no entanto, é a inacessibilidade destes ensaios para a comunidade em geral, e o limitado conhecimento dos mecanismos subjacentes reconhecimento dos outros. Em outras palavras, com base na emissão de um sinal por um indivíduo e o seu reconhecimento por outro ainda está mal compreendida 18. Em contraste, as moscas também exibem um comportamento simples, evitação social, onde os indivíduos se afastar de um sinal emitido por moscas estressados: o D. Melanogaster Estresse Odorant ou DSO 19. Em um ensaio de elevado débito, este comportamento pode ser quantificada como a prevenção de um sinal de tensão emitida por outros, moscas ou evitação social 19. As moscas são colocadas num aparelho de labirinto em T e dada a opção de evitar um frasco contendo DSO. Utilizando este ensaio, CO 2 mostrou ser um componente do DSO, e parte do circuito neural necessário para responder ao CO 2 foi dissecada 19. O ensaio de evitação social aqui apresentada é semelhante conceitualmente para os ensaios comportamentais simples desenvolvido no laboratório de Seymour Benzer que permitiram gerações de pesquisadores para dissecar comportamentos complexos 20. Análise de evitação social pode ser levada a cabo de forma eficaz em termos de custos, utilizando o ensaio de labirinto em T, para permitir um estudo mais generalizada do comportamento social. Por exemplo, com este ensaio que recentemente demonstrou que diferentes riscos genéticos para autismo têm efeitos contrastantes em b sociaisensaios ehavior. Mutantes para um gene candidato para o autismo - neurobeachin 21,22 - deficiências presentes tanto no espaço social (descritos em outros 23) e evitação social 24. Sinalização dopaminérgica anormal também é proposto para desempenhar um papel na etiologia de autismo em humanos 25,26. Em contraste com os resultados obtidos com neurobeachin, descobrimos que o desempenho evitação social não foi afectada por um aumento ou diminuição dos níveis de Drosophila vesicular da monoamina transportador (VMAT) em células dopaminérgicas, embora espaço social foi directamente correlacionada com estes níveis de VMAT 27. Os resultados contrastantes obtidos com neurobeachin e VMAT ressaltam a possibilidade de identificar várias formas de comportamento anti-social e, assim, os diferentes circuitos neurais subjacentes modulando a resposta para os outros.

<table><tbody><tr><td>Stereo Zoom Microscope&amp;nbsp;</td><td>Nikon&amp;nbsp;&amp;nbsp;</td><td>SMZ-645</td><td>Any other standard scope for fly handling would work</td></tr><tr><td>Small paint brushes&amp;nbsp;</td><td></td><td></td><td>for pushing flies</td></tr><tr><td>Porous Polyethylene, 12&quot; x 12&quot; Sheet</td><td>Flystuff - Genesee</td><td>46-100</td><td>http://www.flystuff.com/ProductInfo.php?productID=46-100&lt;br/&gt; Porous Plastic sheet for the cold anesthesia box</td></tr><tr><td>Mini-Alarm Timer/Stopwatch</td><td></td><td></td><td></td></tr><tr><td>Sharpie pens</td><td></td><td></td><td></td></tr><tr><td>Adhesive Tape</td><td></td><td></td><td></td></tr><tr><td>Mini vortex</td><td>Fisher</td><td>14-955-151&amp;nbsp;</td><td>http://www.fishersci.com/ecomm/servlet/itemdetail&lt;br/&gt; For mechanical agitation of the flies - any vortex would work.</td></tr><tr><td>Corning Life Sciences DL No.:352017, Falcon test tube; round bottom; disposable; no closure, 14 ml; 17 x 100 mm</td><td>Fisher</td><td>14-959-8 &amp;nbsp;&amp;nbsp;</td><td>http://www.fishersci.com/ecomm/servlet/itemdetail?storeId=10652&amp;amp;langId=-1&amp;amp;catalogId=29104&amp;amp;productId=2771811&amp;amp;distype=0&amp;amp;highlightProductsItemsFlag=Y&amp;amp;fromSearch=1&amp;amp;searchType=PROD&amp;amp;hasPromo=0&lt;br/&gt; These snap in place in the in-house made T-maze and counter-current apparatus (see text)</td></tr><tr><td>cotton balls</td><td></td><td></td><td>to close the vials after the experiment.</td></tr><tr><td>trifold board and white bench cover</td><td></td><td></td><td>to provide a white background, and a homogeneous light.</td></tr><tr><td>white bench cover</td><td></td><td></td><td></td></tr><tr><td>pounding pad</td><td></td><td></td><td>any mouse pad works.</td></tr><tr><td>large black cloth</td><td></td><td></td><td>to cover the counter-current apparatus in phototaxis response.</td></tr><tr><td>cool-white light&amp;nbsp;</td><td>Home Depot</td><td>1000516563</td><td>http://www.homedepot.ca/product/illume-26-fluorescent-plug-in-linear/911423&lt;br/&gt; any similar linear light with fluorescent light bulb cool-white at 13-15 W would work</td></tr></tbody></table>

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.

O ensaio de evitação social é um ensaio robusto quantificar a capacidade de Drosophila melanogaster para reconhecer um sinal de tensão (DSO) emitidos por outras moscas, e avaliando assim um aspecto de interacções sociais. O ensaio é realizado utilizando um aparelho comumente usado em vários ensaios de comportamento conhecido como T-labirinto, que apresentam as moscas com uma escolha entre duas opções diferentes - a esquerda ou direita 19,28-31. Neste caso, a eficiência com que as moscas ev...

Watch this Scientific Journal Video about Straightforward Assay for Quantification of Social Avoidance in Drosophila melanogaster at JoVE.com

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 ...

straightforward-assay-for-quantification-social-avoidance-drosophila

Research

JoVE Journal

Neuroscience

Straightforward Assay for Quantification of Social Avoidance in Drosophila melanogaster

11.2K visualizações.  Yale University. 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.

Vídeo: Straightforward Assay for Quantification of Social Avoidance in Drosophila melanogaster

Assaying Locomotor, Learning, and Memory Deficits in Drosophila Models of Neurodegeneration

Advances in genetic methods have enabled the study of genes involved in human neurodegenerative diseases using Drosophila as a model system1. Most of these diseases, including Alzheimer's, Parkinson's and Huntington's disease are characterized by age-dependent deterioration in learning and memory functions and movement coordination2. Here we use behavioral assays, including the negative geotaxis assay3 and the aversive phototaxic suppression assay (APS assay)4,5, to show that some of the behavior characteristics associated with human neurodegeneration can be recapitulated in flies. In the negative geotaxis assay, the natural tendency of flies to move against gravity when agitated is utilized to study genes or conditions that may hinder locomotor capacities. In the APS assay, the learning and memory functions are tested in positively-phototactic flies trained to associate light with aversive bitter taste and hence avoid this otherwise natural tendency to move toward light. Testing these trained flies 6 hours post-training is used to assess memory functions. Using these assays, the contribution of any genetic or environmental factors toward developing neurodegeneration can be easily studied in flies.

Assaying Locomotor, Learning, and Memory Deficits in Drosophila Models of Neurodegeneration

Behavior assays for measuring locomotor functions, learning, and memory abilities in Drosophila.

Locomotor ensaio, Aprendizagem e Memória Os déficits no Drosophila Modelos de neurodegeneração

Advances in genetic methods have enabled the study of genes involved in human neurodegenerative diseases using Drosophila as a model system1. Most of ...

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 ...

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 ...

Drosophila Passive Avoidance Behavior as a New Paradigm to Study Associative Aversive Learning

This protocol describes a new paradigm for analyzing aversive associative learning in adult flies (Drosophila melanogaster). The paradigm is analogous to passive avoidance behavior in laboratory rodents in which animals learn to avoid a compartment where they have previously received an electric shock. The assay takes advantage of negative geotaxis in flies, which manifests as an urge to climb up when they are placed on a vertical surface. The setup consists of vertically oriented upper and lower compartments. On the first trial, a fly is placed into a lower compartment from where it usually exits&#160;within 3-15 s, and steps into the upper compartment where it receives an electric shock. During the second trial, 24 h later, the latency is significantly increased. At the same time, the number of shocks is decreased compared to the first trial, indicating that flies formed long-term memory about the upper compartment. The recordings of latencies and number of shocks could be performed with a tally counter and a stopwatch or with an Arduino-based simple device. To illustrate how the assay can be used, the passive avoidance behavior of D. melanogaster and D. simulans male and female were characterized here. Comparison of latencies and number of shocks revealed that both D. melanogaster and D. simulans flies efficiently learned the passive avoidance behavior. No statistical differences were observed between male and female flies. However, males were a little faster while entering the upper compartment on the first trial, while females received a slightly higher number of shocks in every retention trial. The Western diet (WD) significantly impaired learning and memory in male flies while flight exercise counterbalanced this effect. Taken together, the passive avoidance behavior in flies offers a simple and reproducible assay that could be used for studying basic mechanisms of learning and memory.

Drosophila Passive Avoidance Behavior as a New Paradigm to Study Associative Aversive Learning

This work describes a simple behavioral paradigm that allows the analysis of aversive associative learning in adult fruit flies. The method is based on suppressing the innate negative geotaxis behavior due to the association formed between a specific environmental context and an electric shock.

Drosófila Comportamento de evasão passiva como um novo paradigma para estudar aprendizagem aversiva associativa

This protocol describes a new paradigm for analyzing aversive associative learning in adult flies (Drosophila melanogaster). The paradigm is analogous to ...

Comportamento

A Simple Technique to Assay Locomotor Activity in Drosophila

Drosophila melanogaster is an ideal model organism for studying various diseases due to its abundance of advanced genetic manipulation techniques and diverse behavioral features. Identifying behavioral deficiency in animal models is a crucial measure of disease severity, for example, in neurodegenerative diseases where patients often experience impairments in motor function. However, with the availability of various systems to track and assess motor deficits in fly models, such as drug-treated or transgenic individuals, an economical and user-friendly system for precise evaluation from multiple angles is still lacking. A method based on the AnimalTracker application programming interface (API) is developed here, which is compatible with the Fiji image processing program, to systematically evaluate the movement activities of both adult and larval individuals from recorded video, thus allowing for the analysis of their tracking behavior. This method requires only a high-definition camera and a computer peripheral hardware integration to record and analyze behavior, making it an affordable and effective approach for screening fly models with transgenic or environmental behavioral deficiencies. Examples of behavioral tests using pharmacologically treated flies are given to show how the techniques can detect behavioral changes in both adult flies and larvae in a highly repeatable manner.

A Simple Technique to Assay Locomotor Activity in Drosophila

The present protocol assesses the locomotor activity of Drosophila by tracking and analyzing the movement of flies in a hand-made arena using open-source software Fiji, compatible with plugins to segment pixels of each frame based on high-definition video recording to calculate parameters of speed, distance, etc.

Uma técnica simples para avaliar a atividade locomotora em Drosophila

Drosophila melanogaster is an ideal model organism for studying various diseases due to its abundance of advanced genetic manipulation techniques and ...

Conditions Affecting Social Space in Drosophila melanogaster

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

Conditions Affecting Social Space in Drosophila melanogaster

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

Condições que afetam Espaço Social em<em&gt; Drosophila melanogaster</em

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

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 Ensaio para examinar Preferências de postura de Pessoa<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. ...

Drosófila Modelos para traços genéticos e comportamentais do transtorno do espectro do autismo

Paradigms for Behavioral Assessment in Drosophila Model of Autism Spectrum Disorder

Autism Spectrum Disorder (ASD) encompasses a heterogeneous group of neurodevelopmental disorders with common behavioral symptoms including deficits in social interaction and ability for communication, enhanced restricted or repetitive behaviors, and also, in some cases, learning disability and motor deficit. Drosophila has served as an unparalleled model organism for modeling a great number of human diseases. As many genes have been implicated in ASD, fruit flies have emerged as a powerful and efficient way to test the genes putatively involved with the disorder. As hundreds of genes, with varied functional roles, are implicated in ASD, a single genetic fly model of ASD is unfeasible; instead, individual genetic mutants, gene knockdowns, or overexpression-based studies of the fly homologs of ASD-associated genes are the common means for gaining insight regarding molecular pathways underlying these gene products. A host of behavioral techniques are available in Drosophila which provide easy readout of deficits in specific behavioral components. Social space assay and aggression and courtship assays in flies have been shown to be useful in assessing defects in social interaction or communication. Grooming behavior in flies is an excellent readout of repetitive behavior. Habituation assay is used in flies to estimate the ability for habituation learning, which is found to be affected in some ASD patients. A combination of these behavioral paradigms can be utilized to make a thorough assessment of the human ASD-like disease state in flies. Using Fmr1 mutant flies, recapitulating Fragile-X syndrome in humans, and POGZ-homolog row knockdown in fly neurons, we have shown quantifiable deficits in social spacing, aggression, courtship behavior, grooming behavior, and habituation. These behavioral paradigms are demonstrated here in their simplest and straightforward forms with an assumption that it would facilitate their widespread use for research on ASD and other neurodevelopmental disorders in fly models.

Autor em destaque: Explorando os sintomas do transtorno do espectro do autismo em moscas-das-frutas - modelos genéticos e testes comportamentais

Autism Spectrum Disorder (ASD) is associated with impaired social and communicative behavior and the emergence of repetitive behavior. For studying the interrelation between ASD genes and behavioral deficits in the Drosophila model, five behavioral paradigms are described in this paper for assaying social spacing, aggression, courtship, grooming, and habituation behavior.

Paradigmas para Avaliação Comportamental no Modelo de Drosophila de Transtorno do Espectro do Autismo

Autism Spectrum Disorder (ASD) encompasses a heterogeneous group of neurodevelopmental disorders with common behavioral symptoms including deficits in ...

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