This work was supported by NIH P20GM103643 to I. Meng.

1. Preparação de larvas <ol><li> Para analisar uma larva desejado para locomoção ou posição preferência, crescer larvas sob condições padrão para a idade desejada para ensaiar 10 utilizando alimentos mosca padrão 11. </li><li> Faça um filtro de malha esticando silk-screen grau de nylon da malha ao longo de um funil. Fixe a malha no pescoço funil com elástico. Colocar o funil num copo. </li><li> Para a coleta de larvas para análise, colher uma espátula cheia de alimentos que contenham larvas de forrageamento do frasco de cultura e lavar com água da torneira RT sobre o filtro de malha, coletando larvas indivíduo diretamente da malha com um pincel 10. </li></ol> 2. Preparação de tubos de ensaio de <ol><li> Para se preparar para tapar os tubos de ensaio, ferva cuidadosamente um gel de agar 4% e despeje em uma placa de Petri a uma profundidade de 1,5 cm. Isto irá fornecer uma tomada para um ou outro lado do tubo quando animal é fechado. A solução a 4% é suficientemente denso para evitar larvas a partir de penetrating as fichas. </li><li> Certifique-se de que os tubos são limpos e claros antes da inserção de larva. Se não estiverem, isto pode bloquear movimentos de ser gravada. </li><li> Para garantir larvas tem umidade suficiente para o movimento, prepare um frasco contendo uma pequena quantidade de água quente. Inverta a garrafa, cuidadosamente orientar a tomada em direção a uma pia e aperte suavemente para expulsar a água do tubo de coleta. </li><li> Insira a saída da garrafa no tubo de ensaio. Espremer o frasco para fornecer vapor de água dentro do tubo de ensaio até uma fina camada de condensação aparece na parede do tubo. Vídeo 1 mostra uma larva movendo-se em um tubo com uma quantidade apropriada de humidade. </li><li> Para selar a larva no tubo, remover o gel de 1,5 centímetros de espessura de agar de placa de Petri e colocar sobre uma superfície de rede para permitir o fluxo de ar sob o ágar de modo que um tampão de agar pode ser inserido dentro do tubo. Pressionar uma das extremidades do tubo de ensaio para o gel duas vezes, para inserir dois tampões de gel em uma extremidade. </li><li> Com um paintbRush, colocar uma larva no interior do tubo, aproximadamente, 1,5 polegadas de profundidade e selar o tubo ao pressionar a extremidade mais próxima da larva no gel de agar. A pressão resultante irá forçar para fora o segundo bujão de gel de agar a partir da extremidade oposta e selar o animal dentro do tubo. </li><li> Colocar os tubos no dispositivo MB5 multi-Beam Drosophila Activity Monitor (DAM), e ajustar as posições das câmaras de ar de modo a que os animais não pode mover-se para além do alcance dos sensores. </li><li> Para garantir que os tubos de ensaio não deslizar para fora da grelha de leitura de infravermelho do dispositivo durante o transporte, mantenha o tubo no lugar fixando um anel de massa de vidraceiro em torno de cada tubo onde contacta o dispositivo de gravação. </li></ol> 3. Medição Atividade <ol><li> Atividade recorde em uma incubadora para evitar leituras imprecisas que possam ocorrer devido a sombras, luzes fluorescentes ou a variações de temperatura no laboratório. Ver Figura 5 para uma disposição sugerida do sistema. </li><li> Situado numa incubadora a 20 ° C (ver Figura 1). Para evitar falsos gravações devido a interferência de fontes de luz incandescentes durante a gravação, desligar as luzes fluorescentes incubadora e usar uma fonte de luz LED separado durante a gravação na incubadora. Realizar um julgamento sem quaisquer animais para assegurar que as condições de luz não aberrante acionar sensores. Não deve haver dados de movimento gravados após este teste. </li><li> Permitir que as larvas se adapte à as definições incubadora a 20 ° C durante 5 minutos antes do início do ensaio. </li><li> Para definir o sistema de DAM para intervalos de gravação desejados (por exemplo, 1 min), garantir DAM software de gravação do sistema é baixado para sediar computador 12 e abra o arquivo DAM sistema antes de conectar a câmara de gravação para a interface PSIU. </li><li> Para definir a freqüência de gravação desejada na qual os dados serão salvos, selecione Preferências e, em seguida, clique em acima ou abaixo da opção de intervalo de leitura para selecionar diferentes intervalos de tempo em que os dadosvai ser armazenado. Por exemplo, seleccionar os intervalos de tempo de leitura 1 segundo a 1 hora. </li><li> Para selecionar parâmetro variável para gravação de dados, escolha preferências e selecione as caixas correspondentes em tipo de dados de saída (contagens, movimentos, posições e habitar, ver Tabela 1). Cada parâmetro proporciona uma única análise da actividade das larvas no interior do tubo. </li><li> Para iniciar a gravação, conecte o monitor a Unidade de Alimentação Interface (PSIU) usando um cabo de telefone CAB6. Conecte o PSIU a uma tomada eléctrica. Uma luz verde indica conexão apropriada. </li><li> Conecte o PSIU através de um cabo Universal Serial Bus (USB) a um Macintosh ou PC com Windows para a gravação de dados. Abra o programa DAM SystemMB1v6x no computador para iniciar automaticamente a gravação de dados. </li><li> Após o tempo de coleta desejado estiver completo, selecione Sair e, em seguida Sair agora na tela da atividade DAM; dados serão salvos automaticamente sob DAM de dados do sistema. Tome este arquivo de dados (por exemplo, monitorar 1) e arraste<html> em uma pasta separada. Isto irá armazenar os dados em bruto a partir do software DAM de modo que eles podem ser posteriormente processados. </li></ol> 4. Preparação de dados para Processing (DAM FileScan) <ol><li> Para processar os dados brutos em um formato compreensível abrir o aplicativo DAM FileScan e escolha Selecionar pasta de dados de entrada. Em seguida, escolha a pasta de dados eo arquivo desejado e selecione a opção de digitalização. Por fim, selecione o comprimento bin para o período de leitura desejada. Isto irá organizar os dados brutos em parâmetros definidos pelo operador eo programa irá reportar os dados coletados nesse intervalo particular. </li><li> Selecione o tipo de dados de saída para analisar a definição de movimento desejado (monitor de contagem, movimentos, habitar). De acordo com o menu leituras adicionais, select sum em bin (isto é, se o comprimento do compartimento é maior do que o intervalo de leitura de sistema seleccionado). </li><li> Nomeie o arquivo de forma apropriada e salve. O arquivo será armazenado no mesmo local que a pasta a partir do passo 3.9. </li></ol>_title &quot;&gt; 5. Acessando Dados de Análise <ol><li> A fim de recolher movimentos médios, processar os dados brutos gerados a partir do passo 4. Para gerar uma tabela em um programa de planilha, abra o arquivo que foi salvo anteriormente (passo 4.3), e quando a janela Assistente de importação de texto exibida, selecione acabamento para abrir os arquivos de dados em um formato de planilha. Nota: Dependendo do tipo de dados analisados, a planilha será lido de forma diferente. Tipicamente para medir movimentos do monitor ou contagens, o período de tempo de estudo serão gravados numericamente, enquanto cada tubo leitura individual receberão uma carta designada. Ao visualizar os dados na planilha, colunas KZ representam os slots para tubos de ensaio 16/01, respectivamente. As linhas representam os pontos de dados coletados. </li><li> Por exemplo, se move foram recolhidos durante 20 min em intervalos de 1 minuto, em média, as linhas 20 para calcular um número médio de movimentos / minuto e outras medições. </li></ol>

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(86), e51449 (2014).</li><li>Troncoso, B., Godoy-Herrera, R., Waldo, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+development+of+larval+movement+patterns+in+Drosophila).">The development of larval movement patterns in Drosophila).</a> Heredity. 58 (1), 321-329 (1987).</li><li>Helfrich, C., Engelmann, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Circadian+rhythm+of+the+locomotor+activity+in+Drosophila.+melanogaster.and+its+mutants+&#8216;sine+oculis&#8217;+and+&#8216;small+optic+lobes.">Circadian rhythm of the locomotor activity in Drosophila. melanogaster.and its mutants &#8216;sine oculis&#8217; and &#8216;small optic lobes.</a> Physiological Entomology. 8 (3), 257-272 (1983).</li><li>Johnson, A. W., Carder, W. 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The authors declare that they have no competing financial interests.

Actividade de larvas de Drosophila é influenciada por uma variedade de factores, incluindo o genótipo 8, 13 anos de idade e 2 temperatura ambiente. Embora os métodos videográficos poderosos, capazes de análise altamente detalhadas foram desenvolvidas por aqueles que estudam locomoção 5, este nível de detalhe pode ser supérflua para aqueles que desejam para determinar os parâmetros básicos da atividade. O método descrito aqui utiliza um dispositivo que est?...

The use of Drosophila as a genetic tool has transformed scientific knowledge of biological systems. Drosophila larvae have been used in a variety of studies including nociception1, development2 and as a model for the study of human disease genes3. Drosophila activity encompasses a range of behaviors that vary under different conditions including temperatures2, exposure to drugs4 and amongst different genotypes. Yet, despite the significant use of the larva as a model organism, a simple, standardized method to analyze larval activity has not been available. Presently, many larval locomotion studies employ sophisticated video analysis software5. While powerful, the complexity of such automated tools may discourage labs that are not already equipped to study locomotion from including analysis of informative activity parameters in their studies. In other current non-automated methods, such as the grid crawling analysis, motion is scored by a human observer, which introduces the possibility of subjectivity and limits throughput to one animal at a time6-7. A similar study used a 5-lane crawling assay, which measured the time it took larvae to travel a certain linear distance8. In such non-automated assays, displacement is measured but this does not account for non-linear travel between the start and end points. As discussed below, the method described here accounts for more of the actual larval movement, is objective, easy to operate, and offers robust throughput.
To easily study larval activity behavior without the compromise of accuracy, efficiency, or cost, this method employs the TriKinetics Drosophila Activity Monitor (DAM), a device often used to study adult activity. Using one device to study both adult and larval activity is cost-effective, and allows direct comparison of motion by animals at these two life stages. The system, featuring the manufacturer&#8217;s highest level of resolution, makes use of 17 infrared detection beams per assay tube, which record larval activity when sensor beams are broken within the 16 individual tubes. The system then automatically saves recorded information to a computer, making it available for manipulation with basic graphing software. The data obtained represents the beams that were broken by individual larvae (which can be converted into a rate), movement when a larva stays within a detector beam and the position of the animals within the assay chambers during a recording period (allowing one to calculate position preference). The system is efficient and relatively simple to operate, and brings highly reproducible basic activity analysis within the reach of any laboratory studying Drosophila larvae.
To demonstrate the power of this assay, data are presented that show its use to verify differences in activity resulting from varying ambient temperatures, as well as through the comparison of a mutant previously described as hypoactive (iav1)9 with a widely analyzed control (w1118).

<table><tbody><tr><td>Drosophila Activity Monitor, Multibeam, 16 tubes, including wires</td><td>TriKinetics Inc.&amp;nbsp;</td><td>MB5</td><td></td></tr><tr><td>Power Supply Interface for Activity Monitor&amp;nbsp;</td><td>TriKinetics Inc.&amp;nbsp;</td><td>PSIU24</td><td></td></tr><tr><td>Glass 80 x 5 mm tubes for Activity Monitor (100)</td><td>TriKinetics Inc.&amp;nbsp;</td><td>PGT 5x80</td><td></td></tr><tr><td>DAMsystemMB1v6x Data Acquisitions Software for Macintonsh OSX (Intel)</td><td>&lt;a href=&quot;http://www.trikinetics.com/&quot;&gt;www.trikinetics.com&lt;/a&gt;</td><td></td><td>free download</td></tr><tr><td>DAMFileScan 108x software for Macintosh</td><td>&lt;a href=&quot;http://www.trikinetics.com/&quot;&gt;www.trikinetics.com&lt;/a&gt;</td><td></td><td>free download</td></tr><tr><td>USB software (PSIUdrivers.zip).</td><td>&lt;a href=&quot;http://www.trikinetics.com/&quot;&gt;www.trikinetics.com&lt;/a&gt;</td><td></td><td>free download</td></tr><tr><td>DAMSystem Notes 308</td><td>&lt;a href=&quot;http://www.trikinetics.com/&quot;&gt;www.trikinetics.com&lt;/a&gt;</td><td></td><td>free download</td></tr><tr><td>Zeiss Stemi 2000C- Stereo Microscope</td><td>Spectra Services</td><td>SP-STEMI2000C-BS</td><td></td></tr><tr><td>Carbon Dioxide</td><td>Maine Oxy</td><td></td><td>anaesthesia</td></tr><tr><td>Fly Pad</td><td>Genesee</td><td>59-114</td><td>surface for sorting anaesthetized flies</td></tr><tr><td>Small paint brush&amp;nbsp;</td><td>Winsor &amp;amp; Newton</td><td>#2 ROUND</td><td>or similar, used for sorting anaesthetized flies</td></tr><tr><td>Silk Screen Printing Mesh (160)</td><td>msj-gallery.com</td><td>SM160W63-3YD</td><td>pore sized used in this protocol was ~ 0.1 mm</td></tr><tr><td>Tegosept</td><td>Genesee</td><td>20-258</td><td>preservative</td></tr><tr><td>Ethanol (190proof)</td><td>Pharmco</td><td>111000190</td><td>used to dissolve Tegosept</td></tr><tr><td>6 oz Square Bottom Bottle (PP)</td><td>Genesee</td><td>32-130</td><td></td></tr><tr><td>&quot;Flugs&quot; for Plastic Fly bottles</td><td>Genesee</td><td>49-100</td><td></td></tr><tr><td>Drosophila Vials, Wide (PS)</td><td>Genesee</td><td>32-117</td><td></td></tr><tr><td>Flugs for wide plastic vials</td><td>Genesee</td><td>49-101</td><td></td></tr><tr><td>Yellow Degerminated Corn Meal</td><td>Gold Medal</td><td></td><td></td></tr><tr><td>Drosophila agar</td><td>LabScientific</td><td>FLY 8020</td><td></td></tr><tr><td>Baker&#039;s Yeast - Red Star</td><td>King Arthur Flour</td><td>1270</td><td></td></tr><tr><td>Granulated Sugar - Extra Fine</td><td>Domino</td><td></td><td></td></tr></tbody></table>

measurement of larval activity in the drosophila activity monitor

Drosophila larvae are used in many behavioral studies, yet a simple device for measuring basic parameters of larval activity has not been available. This protocol repurposes an instrument often used to measure adult activity, the TriKinetics Drosophila activity monitor (MB5 Multi-Beam Activity Monitor) to study larval activity. The instrument can monitor the movements of animals in 16 individual 8 cm glass assay tubes, using 17 infrared detection beams per tube. Logging software automatically saves data to a computer, recording parameters such as number of moves, times sensors were triggered, and animals&#8217; positions within the tubes. The data can then be analyzed to represent overall locomotion and/or position preference as well as other measurements. All data are easily accessible and compatible with basic graphing and data manipulation software. This protocol will discuss how to use the apparatus, how to operate the software and how to run a larval activity assay from start to finish.

A Figura 1 mostra os resultados de um estudo de resposta à temperatura de controlo de larvas de terceiro instar, W 1118, utilizando o dispositivo de monitorização, para detectar diferenças na locomoção das larvas em sete diferentes temperaturas. As larvas foram lavadas e colocadas no dispositivo de actividade DAM como descrito acima, e colocado numa incubadora à temperatura desejada. O aparelho foi então deixados a aclimatar ao ambiente durante 5 min antes da gravação começou. Cada...

Watch this Scientific Journal Video about Measurement of Larval Activity in the Drosophila Activity Monitor at JoVE.com

Measurement of Larval Activity in the Drosophila Activity Monitor

This report describes a method for measuring Drosophila larval activity using the TriKinetics Drosophila Activity Monitor. The device employs infrared beams to detect movements of up to 16 individual animals. Data can be analyzed to represent motion parameters including rates and the positions of the animals within the assay chambers.

Medição da Actividade larval no<em&gt; Drosophila</em&gt; Monitor de Atividade

Drosophila larvae are used in many behavioral studies, yet a simple device for measuring basic parameters of larval activity has not been available. This ...

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Biology

Measurement of Larval Activity in the Drosophila Activity Monitor

9.3K visualizações.  University of New England. This report describes a method for measuring Drosophila larval activity using the TriKinetics Drosophila Activity Monitor. The device employs infrared beams to detect movements of up to 16 individual animals. Data can be analyzed to represent motion parameters including rates and the positions of the animals within the assay chambers.

Vídeo: Measurement of Larval Activity in the Drosophila Activity Monitor

Assaying Locomotor Activity to Study Circadian Rhythms and Sleep Parameters in Drosophila

Most life forms exhibit daily rhythms in cellular, physiological and behavioral phenomena that are driven by endogenous circadian (&equiv;24 hr) pacemakers or clocks. Malfunctions in the human circadian system are associated with numerous diseases or disorders. Much progress towards our understanding of the mechanisms underlying circadian rhythms has emerged from genetic screens whereby an easily measured behavioral rhythm is used as a read-out of clock function. Studies using Drosophila have made seminal contributions to our understanding of the cellular and biochemical bases underlying circadian rhythms. The standard circadian behavioral read-out measured in Drosophila is locomotor activity. In general, the monitoring system involves specially designed devices that can measure the locomotor movement of Drosophila. These devices are housed in environmentally controlled incubators located in a darkroom and are based on using the interruption of a beam of infrared light to record the locomotor activity of individual flies contained inside small tubes. When measured over many days, Drosophila exhibit daily cycles of activity and inactivity, a behavioral rhythm that is governed by the animal's endogenous circadian system. The overall procedure has been simplified with the advent of commercially available locomotor activity monitoring devices and the development of software programs for data analysis. We use the system from Trikinetics Inc., which is the procedure described here and is currently the most popular system used worldwide. More recently, the same monitoring devices have been used to study sleep behavior in Drosophila. Because the daily wake-sleep cycles of many flies can be measured simultaneously and only 1 to 2 weeks worth of continuous locomotor activity data is usually sufficient, this system is ideal for large-scale screens to identify Drosophila manifesting altered circadian or sleep properties.

Assaying Locomotor Activity to Study Circadian Rhythms and Sleep Parameters in Drosophila

We describe procedures for recording daily locomotor activity rhythms of Drosophila and subsequent data analysis. Locomotor activity rhythms are a reliable behavioral output of animal circadian clocks and are used as the standard readout of clock function when studying circadian mutants or examining how the environment regulates the circadian system.

Assaying atividade locomotora para estudar ritmos circadianos e sono Parâmetros em Drosophila</em

Most life forms exhibit daily rhythms in cellular, physiological and behavioral phenomena that are driven by endogenous circadian (&equiv;24 hr) ...

Neurociência

Measuring Exercise Levels in Drosophila melanogaster Using the Rotating Exercise Quantification System (REQS)

Drosophila melanogaster is a new model organism for studies in exercise biology. To date, two main exercise systems, the Power Tower and the Treadwheel have been described. However, a method to measure the amount of additional animal activity induced through the exercise treatment has been lacking. The Rotating Exercise Quantification System (REQS) fills this need, providing a measure of animal activity for animals experiencing rotational exercise. This protocol details how to use the REQS to assess animal activity during rotational exercise and illustrates the type of data that can be generated. Here, we demonstrate how the REQS is used to measure sex- and strain-specific differences in exercise induced activity. The REQS can also be used to evaluate the impact of various other experimental parameters such as age, diet, or population size on exercise induced activity. In addition, it can be used to compare the efficacy of different exercise training protocols. Importantly, it provides an opportunity to standardize exercise treatments between strains, allowing the researcher to achieve equal amounts of activity between groups if needed. Thus, the REQS is a notable new resource for exercise biologists working with the Drosophila model system and complements existing exercise systems.

Measuring Exercise Levels in Drosophila melanogaster Using the Rotating Exercise Quantification System REQS

The Rotating Exercise Quantification System (REQS) can induce exercise in Drosophila melanogaster through rotation while simultaneously measuring the amount of activity performed by the animals. Here, we present a point-by-point protocol detailing how to measure activity levels of animals experiencing rotational exercise treatments using the REQS.

Medição de níveis de exercício em Drosophila melanogaster , usando o sistema de quantificação de exercício de giro (REQS)

Drosophila melanogaster is a new model organism for studies in exercise biology. To date, two main exercise systems, the Power Tower and the Treadwheel ...

Genética

A Computational Method to Quantify Fly Circadian Activity

In most animals and plants, circadian clocks orchestrate behavioral and molecular processes and synchronize them to the daily light-dark cycle. Fundamental mechanisms that underlie this temporal control are widely studied using the fruit fly Drosophila melanogaster as a model organism. In flies, the clock is typically studied by analyzing multiday locomotor recording. Such a recording shows a complex bimodal pattern with two peaks of activity: a morning peak that happens around dawn, and an evening peak that happens around dusk. These two peaks together form a waveform that is very different from sinusoidal oscillations observed in clock genes, suggesting that mechanisms in addition to the clock have profound effects in producing the observed patterns in behavioral data. Here we provide instructions on using a recently developed computational method that mathematically describes temporal patterns in fly activity. The method fits activity data with a model waveform that consists of four exponential terms and nine independent parameters that fully describe the shape and size of the morning and evening peaks of activity. The extracted parameters can help elucidate the kinetic mechanisms of substrates that underlie the commonly observed bimodal activity patterns in fly locomotor rhythms.

A method to quantify the main temporal features seen in fly circadian locomotor rhythms is presented. The quantification is achieved by fitting fly activity with a multi-parametric model waveform. The model parameters describe the shape and size of the morning and evening peaks of daily activity.

Um método computacional para quantificar a atividade Circadian voar

In most animals and plants, circadian clocks orchestrate behavioral and molecular processes and synchronize them to the daily light-dark cycle. ...

Comportamento

Quantitative Measurement of the Immune Response and Sleep in Drosophila

A complex interaction between the immune response and host behavior has been described in a wide range of species. Excess sleep, in particular, is known to occur as a response to infection in mammals 1 and has also recently been described in Drosophila melanogaster2. It is generally accepted that sleep is beneficial to the host during an infection and that it is important for the maintenance of a robust immune system3,4. However, experimental evidence that supports this hypothesis is limited4, and the function of excess sleep during an immune response remains unclear. We have used a multidisciplinary approach to address this complex problem, and have conducted studies in the simple genetic model system, the fruitfly Drosophila melanogaster. We use a standard assay for measuring locomotor behavior and sleep in flies, and demonstrate how this assay is used to measure behavior in flies infected with a pathogenic strain of bacteria. This assay is also useful for monitoring the duration of survival in individual flies during an infection. Additional measures of immune function include the ability of flies to clear an infection and the activation of NF&kappa;B, a key transcription factor that is central to the innate immune response in Drosophila. Both survival outcome and bacterial clearance during infection together are indicators of resistance and tolerance to infection. Resistance refers to the ability of flies to clear an infection, while tolerance is defined as the ability of the host to limit damage from an infection and thereby survive despite high levels of pathogen within the system5. Real-time monitoring of NF&kappa;B activity during infection provides insight into a molecular mechanism of survival during infection. The use of Drosophila in these straightforward assays facilitates the genetic and molecular analyses of sleep and the immune response and how these two complex systems are reciprocally influenced.

Quantitative Measurement of the Immune Response and Sleep in Drosophila

To understand a link between the immune response and behavior, we describe a method to measure locomotor behavior in Drosophila during bacterial infection as well as the ability of flies to mount an immune response by monitoring survival, bacterial load, and real-time activity of a key regulator of innate immunity, NF&kappa;B.

Medição quantitativa da resposta imune e do sono em<em&gt; Drosophila</em

A complex interaction between the immune response and  host behavior has been described in a wide range of species. Excess sleep, in  particular, is known ...

Imunologia e Infecção

High-Resolution Video Tracking of Locomotion in Adult Drosophila Melanogaster

Flies provide an important model for studying complex behavior due to the plethora of genetic tools available to researchers in this field. Studying locomotor behavior in Drosophila melanogaster relies on the ability to be able to quantify changes in motion during or in response to a given task. For this reason, a high-resolution video tracking system, such as the one we describe in this paper, is a valuable tool for measuring locomotion in real-time. Our protocol involves the use of an initial air pulse to break the flies momentum, followed by a thirty second filming period in a square chamber. A tracking program is then used to calculate the instantaneous speed of each fly within the chamber in 10 msec increments. Analysis software then compiles this data, and outputs a variety of parameters such as average speed, max speed, time spent in motion, acceleration, etc. This protocol will discuss proper feeding and management of flies for behavioral tasks, handling flies without anesthetization or immobilization, setting up a controlled environment, and running the assay from start to finish.

The study of complex locomotor behavior in Drosophila melanogaster is dependent upon the ability to quantify changes in a given fly's movement. This article demonstrates how to do this using a high-resolution tracking system.

De alta resolução de vídeo de Rastreamento Locomotion em Adult Drosophila melanogaster

Flies provide an important model for studying complex behavior due to the plethora of genetic tools available to researchers in this field.  Studying ...

Biologia

Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster

Drosophila melanogaster has been used as an excellent model organism to study environmental and genetic manipulations that affect behavior. One such behavior is spontaneous locomotor activity. Here we describe our protocol that utilizes Drosophila population monitors and a tracking system that allows continuous monitoring of the spontaneous locomotor activity of flies for several days at a time. This method is simple, reliable, and objective and can be used to examine the effects of aging, sex, changes in caloric content of food, addition of drugs, or genetic manipulations that mimic human diseases.

Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster

Drosophila melanogaster are useful in studying genetic or environmental manipulations that affect behaviors such as spontaneous locomotor activity.&#160; Here we describe a protocol that utilizes monitors with infrared beams and data analysis software to quantify spontaneous locomotor activity.&#160;

Determinação da atividade locomotora espontânea em<em&gt; Drosophila melanogaster</em

Drosophila melanogaster has been used as an excellent model organism to study environmental and genetic manipulations that affect behavior. One such ...

A Low-cost Method for Analyzing Seizure-like Activity and Movement in Drosophila

Video tracking systems have been used widely to analyze Drosophila melanogaster movement and detect various abnormalities in locomotive behavior. While these systems can provide a wealth of behavioral information, the cost and complexity of these systems can be prohibitive for many labs. We have developed a low-cost assay for measuring locomotive behavior and seizure movement in D. melanogaster. The system uses a web-cam to capture images that can be processed using a combination of inexpensive and free software to track the distance moved, the average velocity of movement and the duration of movement during a specified time-span. To demonstrate the utility of this system, we examined a group of D. melanogaster mutants, the Bang-sensitive (BS) paralytics, which are 3-10 times more susceptible to seizure-like activity (SLA) than wild type flies. Using this novel system, we were able to detect that the BS mutant bang senseless (bss) exhibits lower levels of exploratory locomotion in a novel environment than wild type flies. In addition, the system was used to identify that the drug metformin, which is commonly used to treat type II diabetes, reduces the intensity of SLA in the BS mutants.

A Low-cost Method for Analyzing Seizure-like Activity and Movement in Drosophila

Using a webcam and a combination of free and inexpensive software programs, locomotive patterns in Drosophila melanogaster can be analyzed to detect differences in the speed, distance, and time of various motor activities.

Um método de baixo custo para análise de Apreensão-como atividade e movimento em<em&gt; Drosophila</em

Video tracking systems have been used widely to analyze Drosophila melanogaster movement and detect various abnormalities in locomotive behavior. While ...

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 ...

<meta charset="utf8"></head><body>Medição de tolerância ao calor de alto rendimento em Drosophila usando sistemas DAM2

Heat Tolerance Assays Using the Drosophila Activity Monitor System: A Guide to an Executable Application for Data Analysis

The study of heat tolerance in&#8239;Drosophila melanogaster has been of particular interest to researchers for decades, with a common approach to assessing heat tolerance being to monitor the time to knockdown (TKD) after exposure to an elevated temperature. Classically, flies are housed in individual vials and placed inside a heated water bath. TKD is then monitored manually by researchers. While very well-established, there remain problems of subjectivity and consistent application of a tangible definition of cessation of all movement, including muscular spasms, when implementing these manual assays. We have developed a high-throughput method for automating heat tolerance assays using the TriKinetics Drosophila Activity Monitors (DAM2). To accompany the DAM2 system, we have written a program and created an easy-use executable to automatically read the last time of movement from the activity data generated. This script then writes to a .csv file the time to heat paralysis (TKD) for each fly. Our data show that this automated DAM2 method is consistent and reliable. Meanwhile, activity profiles created from the activity count data are of interest. These activity profiles can be compiled and have the potential to expand heat tolerance assays to include the relatively unstudied behavioral components of heat tolerance. This protocol will describe in detail how to use the DAM2 system and the HoTDAM! software to estimate heat tolerance in D. melanogaster.

<meta charset="utf8"></head><body>Ensaios de tolerância ao calor usando o sistema de monitoramento de atividade de Drosophila : um guia para um aplicativo executável para análise de dados</p

This report describes a method for measuring adult Drosophila melanogaster time to knockdown using a Drosophila Activity Monitor (DAM2) in response to an air conduction heat stressor within an incubator chamber. The DAM2 measures activity by recording individual fly movements as they cross an infrared beam. Data analysis is facilitated by a novel executable file created by the authors.

Ensaios de tolerância ao calor usando o sistema de monitoramento de atividade de Drosophila : um guia para um aplicativo executável para análise de dados

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Ensaios de tolerância ao calor usando o sistema de monitoramento de atividade de Drosophila : um guia para um aplicativo executável para análise de dados