우리는 비행 주식에 대한 블루밍턴 Drosophila 주식 센터 감사드립니다; C. Gahr, JT Alkema 및 S. van Hasselt가 가압 공기 분석법 개발 초기 시도 재스퍼 보스 만 (Jasper Bosman) : 효모 배양에 대한 조언; 원래는 Drosophila 교배 행동의 저속 모니터링을 개발 한 Rezza Azanchi와 Joel Levine입니다. JA Gorter는 Neuroscience Research School BCN / NWO 대학원 프로그램 지원금으로 지원되었습니다. 이 작업은 JC Billeter에 대한 네덜란드 연구 기관 (NWO) (참고 : 821.02.020)이 부분적으로 지원했습니다.

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저자는 공개 할 경쟁적 이익이 없습니다.

이 프로토콜은 짝짓기 커플이 짝짓기 빈도를 결정하는 데 사용하도록 가정 된 환경 신호를 지속적으로 제어하면서 24 시간 동안 짝짓기 행위를 테스트하는 분석을 설명합니다. 배지에 효모가 들어있을 때 가압 공기 시스템을 통해 전달되는 효모 공기에 반응하여 결합 빈도를 높일 수 있습니다 ( 그림 2B ). 또한 짝짓기 빈도의 비슷한 반응은 배지에서 한천, 펩톤 및 아세트산 냄새만을 ...

생식 행동은 일반적으로 남성보다 큰 배우자를 생산하고 개발하는 자손을 키우기위한 조건을 신중하게 선택해야하는 여성에게 특히 높은 에너지 비용을 초래합니다. 에너지 비용 때문에 재생산이 영양 상태와 관련이 있다는 것은 놀라운 일이 아닙니다. 이것은 포유류를 포함한 대부분의 동물에서 마찬가지입니다. 포유 동물은 영양 부족으로 사춘기가 늦어 질 수 있으며 성행위가 식량 제한으로 인해 부정적인 영향을받을 수 있습니다 1 . 유전 모델 유기체 Drosophila melanogaster 의 번식은 영양 상태에 의해서도 영향을받습니다. 식품 휘발성이의 존재에 높은 수준의 남성 코트, 그리고 여성보다 성적 효모, 계란 생산 및 자손의 생존 3, 4, 5의 주요 영양소의 존재에 수용된다. 이진화 보존 된 식품에 대한 생식 적 반응은 환경 친화적 인 식품 이용 가능성을 유 전적으로 다루기 쉽고 시간이 절약되는 유기체에서 성 생식에 연결시키는 메커니즘을 연구 할 기회를 제공합니다. 실제로 D. melanogaster 에서의 연구는 인슐린 경로가 음식과 교미 행동 사이의 연관성에 대한 중요한 조절 자라는 것을 의미합니다 6 . 또한 자체를 정합하는 단계는 암컷의 음식 선호도뿐만 아니라 관련된 화학 감각 뉴런 7, 8, 9를 변경하는 것으로 나타났다. D. melanogaster 에서 음식 단서가 생식 행동에 영향을주는 것은 분명합니다. 이러한 효과는 주로 여성, 특히 이미 5를 짝 지어 준 여성에게 영향을 미치는 것으로 보입니다. 그러나 환경 조건의 급성 영향을 시험하기 위해 여성의 행동에 고전적으로 사용되는 분석법이짝짓기 에피소드 간의 오랜 중단으로 인해 적합하지 않습니다. 클래식 remating 분석에서 처녀 여성 먼저 남성과 짝짓기, 즉시 격리하고 새로운 남성과 함께 24 ~ 48 시간 후에 제공됩니다. 이 고전적 분석법은 여성의 행동과 여성 반응을 수정하는 남성 사정의 구성 요소를 확인하는 데 큰 성공을 거두었습니다 12 , 13 , 14 , 15 , 16 , 17 , 18 . 따라서 여기에서 입증 된 연속 교미 분석은 재생산 행동에 대한 환경 조건의 급성 효과를 연구하는 데 사용할 수있는 고전적 교미 분석법에 추가 된 것입니다. 여기에 설명 된 짝짓기 행동에 대한 연속 분석법을 사용하여 이전에 효모에 노출 된 한 쌍의 파리가24 시간 관찰 기간 5, 19, 20, 21 배 이상 everal, 파리만이 remate 한번 음식 (5)에 노출되지 않는 상태. 이 발견은 D. melanogaster 문헌의 많은 부분에 비추어 수수께끼 일 수 있습니다 . 암컷은 초기 교미 후 며칠 동안 remate하지 않는다는 것을 나타냅니다 (참고 문헌 10 , 11 에서 검토 됨). 그러나 이러한 불일치는 새로운 교미 기회가 제공되기 전에 암컷이 1 일에서 며칠 동안 격리 된 분석 조건에 의해 쉽게 설명 될 수 있습니다. 쌍이이 1 시간의 관측 기간에 짝짓기하지 않으면 암컷은 감수성이없는 것으로 특징 지어진다. 더욱이, 야생 잡힌 파리의 자료에 따르면 암컷에 저장 기관에 4 ~ 6 마리의 수컷이 들어있는 정자가 있다는 것을 감안할 때, 높은 교미 빈도는 놀랄만 한 일이 아니어야한다. 따라서여성 자연스럽게, 23 차례 (22) remate 것을 dicating. 여기에서 우리는 파리가 어떻게 모여지고 환경 조건에 대한 정보를 결합하여 짝짓기 빈도를 조절하는지에 대한이 연속 교배 분석법의 사용을 입증합니다. 이 분석을 통해 유전자 연구를 위해 비교적 많은 수의 교미 부부를 시험하고 휘발성 및 비 휘발성 환경 단서의 영향을 시험 할 수 있습니다. 이 분석은 일반적으로 24 시간 동안 실행되지만 48 시간까지 연장 될 수있어 명암주기 (light-dark, LD)주기와 같은 사이클링 환경 큐를 테스트 할 수 있습니다. 우리는 식품 기질에서 비 휘발성 효모 영양소의 가용성과 함께 가압 공기 시스템 내에서 효모 배양으로부터 휘발성 신호의 영향을 시험함으로써이 분석을 입증합니다. 가압 공기 시스템은 지속적으로 휘발성 큐를식품 기질 및 시험 커플 (그의 교미 행동이 모니터 됨). 효모가 짝짓기에 영향을주는 세부 사항을 더 결정하기 위해, 우리는 효모의 주요 휘발성 화합물, 즉 아세트산 24 를 펩톤 (아미노산)의 형태로 식품 기질의 효모에 상응하는 아미노산 함량과 함께 시험한다 동물성 단백질의 효소 소화에서 유래 된 산). 이 실험들은 함께 D. melanogaster 의 교미 작용에 대한 환경 신호의 영향이이 분석법으로 시험 될 수 있음을 보여줍니다.

<table><tbody><tr><td>&lt;strong&gt;Cabinet&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Stainless steel kitchen cabinet</td><td>Horecaworld</td><td>7412.0105</td><td></td></tr><tr><td>White LEDs</td><td>Lucky Light</td><td>ll-583wc2c-001</td><td>Cold white, 20 mAmp and 2 V</td></tr><tr><td>Red LEDs</td><td>Lucky Ligt</td><td>ll-583vc2c-v1-4da</td><td>Wavelength between 625 nm, 20 mAmp and 6 V</td></tr><tr><td>Resistor</td><td>Royal Ohm</td><td>CFR0W4J0561A50</td><td>560 ohm, 0.25 W, 250 V and 5 % tolerance</td></tr><tr><td>Smartphone light meter app</td><td>Patrick Giudicelli</td><td></td><td>Light/Lux Meter FREE, version 1.1.1</td></tr><tr><td>Power timer</td><td>Alecto</td><td>TS-121</td><td></td></tr><tr><td>Metal brackets</td><td></td><td></td><td>Sharp angle 5 by 5 mm,&amp;nbsp; 2 x 5450 and 1 x 1100 mm long</td></tr><tr><td>Frosted glass plate</td><td></td><td></td><td>1190 x 545 x 5 mm</td></tr><tr><td>Filter paper sheets</td><td>LEE filters</td><td>220</td><td>White frost</td></tr><tr><td>Small fan</td><td>Nanoxia Deep silence</td><td>4260285292828</td><td>80 mm Ultra-Quiet PC Fan, 1200 RPM</td></tr><tr><td>Big fan</td><td>Nanoxia Deep silence</td><td>4260285292910</td><td>120 mm Ultra-Quiet PC Fan, 650-1500 RPM</td></tr><tr><td>Webcam camera</td><td>Logitech</td><td>950270</td><td>B910 HD WEBCAM OEM, Angle: 78-degree, resolution: 5-million-pixel&amp;nbsp;&amp;nbsp;</td></tr><tr><td>Camera software</td><td>DeskShare</td><td></td><td>Security monitor pro</td></tr><tr><td>&lt;strong&gt;Name&lt;/strong&gt;</td><td>&lt;strong&gt;Company&lt;/strong&gt;</td><td>&lt;strong&gt;Catalog Number&lt;/strong&gt;</td><td>&lt;strong&gt;Comments&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;Fly rearing&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Fly rearing bottles</td><td>Flystuff</td><td>32-130</td><td>6oz Drosophila stock bottle</td></tr><tr><td>Flypad</td><td>Flystuff</td><td>59-114</td><td></td></tr><tr><td>Wild-type flies</td><td></td><td></td><td>Canton-S</td></tr><tr><td>Fly rearing vials</td><td>Dominique Dutscher</td><td>789008</td><td>Drosophila tubes narrow 25x95 mm</td></tr><tr><td>Incubator</td><td>Sanyo</td><td>MIR-154</td><td></td></tr><tr><td>Magnetic hot plate</td><td>Heidolph</td><td>505-20000-00</td><td>MR Hei-Standard</td></tr><tr><td>Agar</td><td>Caldic Ingredients B.V.</td><td>010001.26.0</td><td></td></tr><tr><td>Glucose</td><td>Gezond&amp;amp;wel</td><td>1019155</td><td>Dextrose/Druivensuiker</td></tr><tr><td>Sucrose</td><td>Van Gilse</td><td></td><td>Granulated sugar</td></tr><tr><td>Cornmeal</td><td>Flystuff</td><td>62-100</td><td></td></tr><tr><td>Wheat germ</td><td>Gezond&amp;amp;wel</td><td>1017683</td><td></td></tr><tr><td>Soy flour</td><td>Flystuff</td><td>62-115</td><td></td></tr><tr><td>Molasses</td><td>Flystuff</td><td>62-117</td><td></td></tr><tr><td>Active dry yeast</td><td>Red Star</td><td></td><td></td></tr><tr><td>Tegosept</td><td>Flystuff</td><td>20-258</td><td>100%</td></tr><tr><td>Peptone (bacto)</td><td>BD</td><td>211677</td><td></td></tr><tr><td>Acetic Acid</td><td>Merck</td><td>1000631000</td><td>Glacial, 100%</td></tr><tr><td>Small petridish</td><td>Greiner bio-one</td><td>627102</td><td>35 x 10 mm with vents</td></tr><tr><td>Paraffin film</td><td>Bemis NA</td><td></td><td>Parafilm</td></tr><tr><td>&lt;strong&gt;Name&lt;/strong&gt;</td><td>&lt;strong&gt;Company&lt;/strong&gt;</td><td>&lt;strong&gt;Catalog Number&lt;/strong&gt;</td><td>&lt;strong&gt;Comments&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;Yeast and pressurised air set-up&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Big petridish</td><td>Gosselin</td><td>BP140-01</td><td>140 x 20.6 mm</td></tr><tr><td>Ultrapure water</td><td>Millipore corporation</td><td></td><td>MiliQ</td></tr><tr><td>Yeast extract</td><td>BD</td><td>212750</td><td></td></tr><tr><td>Agar (pure)</td><td>BD</td><td>214530</td><td>bacto</td></tr><tr><td>Glucose (0(+)-glucose monohydrate)&amp;nbsp;</td><td>Merck</td><td>18270000004</td><td></td></tr><tr><td>Open caps</td><td>Schott</td><td>29 240 28&amp;nbsp;</td><td>GL45</td></tr><tr><td>Silicone septum</td><td>VWR</td><td>548-0662</td><td></td></tr><tr><td>Barbed bulkhead fittings</td><td>Nalgene</td><td>6149-0002</td><td></td></tr><tr><td>Large PVC tubing</td><td></td><td></td><td>diameter: outer 1.2 cm and inner 0.9 cm</td></tr><tr><td>Small PVC tubing</td><td></td><td></td><td>diameters: outer 0.8 cm and inner 0.5 cm</td></tr><tr><td>15 ml tube</td><td>Falcon</td><td></td><td></td></tr><tr><td>Aquarium pump</td><td>Sera precision</td><td></td><td>Sera air 110 plus, AC 220-240 V, 50/60 Hz, 3 W and pressure &gt;100 mbar</td></tr><tr><td>Activated charcoal</td><td>Superfish</td><td>A8040400</td><td>Norit activated carbon</td></tr><tr><td>Disposible filter unit</td><td>Whatman</td><td>10462100</td><td></td></tr><tr><td>Serological pipettes</td><td>VWR</td><td>612-1600</td><td></td></tr><tr><td>Syringe</td><td>BD Plastipak</td><td>300013</td><td></td></tr><tr><td>Hot glue</td><td>Pattex</td><td></td><td></td></tr><tr><td>Syringe filter</td><td>Whatman</td><td></td><td>FP 30/pore size 0.45 mm CA-S</td></tr><tr><td>&lt;strong&gt;Name&lt;/strong&gt;</td><td>&lt;strong&gt;Company&lt;/strong&gt;</td><td>&lt;strong&gt;Catalog Number&lt;/strong&gt;</td><td>&lt;strong&gt;Comments&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;Analysis&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Statistics software</td><td>R</td><td></td><td>lme4 package</td></tr></tbody></table>

a method to test the effect of environmental cues on mating behavior in drosophila melanogaster

개인의 성행위는 유전자형, 경험 및 환경 조건의 영향을받습니다. 이러한 요인들이 어떻게 상호 작용하여 성적 행동을 조절하는지는 여전히 잘 알려져 있지 않습니다. Drosophila melanogaster 에서는 식량 가용성과 같은 환경 적 단서가 성적 행동을 조절하는 메커니즘을 조사 할 수있는 다루기 쉬운 시스템을 제공하는 교미 활동에 영향을 미칩니다. D. melanogaster에서 , 환경 단서는 종종 화학 감각 기관과 후각 시스템을 통해 감지됩니다. 여기에서는 짝짓기 행동에 환경 화학 신호의 영향을 테스트하는 방법을 제시합니다. 이 분석법은 식품 매체와 짝짓기 커플을 포함하는 작은 짝짓기 경기장으로 구성됩니다. 각 커플의 교미 빈도는 24 시간 동안 지속적으로 모니터링됩니다. 여기서 우리는 가청 공기 시스템을 통해 외부 소스로부터 환경 화합물을 시험 할뿐만 아니라 짝짓기 경기장에서 환경 성분을 직접 조작하는이 분석의 적용 가능성을 제시합니다. 너는가압 에어 시스템은 매우 휘발성 인 화합물의 효과를 테스트하는 데 특히 유용하며, 상대방의 구성 요소를 직접 조작하는 것은 화합물의 존재를 확인하는 데 가치가 있습니다. 이 분석은 유전 적 및 환경 적 단서가 짝짓기 행동과 생식력 및 기타 남성과 여성의 생식 행동에 미치는 영향에 대한 질문에 대답 할 수 있습니다.

이 연속 분석법을 사용하여 실험적 환경 조건 하에서 교미 행동 및 특정 주파수에서의 교미 빈도를 결정할 수 있습니다. 환경 조건을 제어하기 위해 우리는 스테인리스 스틸 주방 캐비닛을 자체 광원 및 확산 장치를 사용하여 테스트 영역으로 변형 시켰습니다.이 캐비닛은 결합 영역의 상단에서 높은 광량과 눈부심을 보장합니다 ( 그림 1A ) . 내부 테스트 영?...

Watch this Scientific Journal Video about A Method to Test the Effect of Environmental Cues on Mating Behavior in Drosophila melanogaster at JoVE.com

A Method to Test the Effect of Environmental Cues on Mating Behavior in Drosophila melanogaster

우리는 초파리 Drosophila melanogaster 에서 짝짓기 행동에 영향을 미치는 환경 및 유전 신호를 분석하는 분석법을 시연합니다.

환경에서 신호의 결합 효과에 미치는 영향을 시험하는 방법<em&gt; Drosophila melanogaster</em

An individual's sexual drive is influenced by genotype, experience and environmental conditions. How these factors interact to modulate sexual behaviors ...

a-method-to-test-effect-environmental-cues-on-mating-behavior

Universidad de Cartagena UDEC

Research

JoVE Journal

Behavior

A Method to Test the Effect of Environmental Cues on Mating Behavior in Drosophila melanogaster

9.0K 조회수.  University of Groningen.  우리는 초파리 Drosophila melanogaster 에서 짝짓기 행동에 영향을 미치는 환경 및 유전 신호를 분석하는 분석법을 시연합니다. 

비디오: A Method to Test the Effect of Environmental Cues on Mating Behavior in Drosophila melanogaster

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.

분석의 방법 Drosophila 행동

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

연구

신경과학

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.

먹이 찾아 돌아다 행동에 대한 단일 비행 분석에<em&gt; 초파리</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 ...

Methods to Test Endocrine Disruption in Drosophila melanogaster

In recent years there has been growing evidence that all organisms and the environment are exposed to hormone-like chemicals, known as endocrine disruptor chemicals (EDCs). These chemicals may alter the normal balance of endocrine systems and lead to adverse effects, as well as an increasing number of hormonal disorders in the human population or disturbed growth and reduced reproduction in the wildlife species. For some EDCs, there are documented health effects and restrictions on their use. However, for most of them, there is still no scientific evidence in this sense. In order to verify potential endocrine effects of a chemical in the full organism, we need to test it in appropriate model systems, as well as in the fruit fly, Drosophila melanogaster. Here we report detailed in vivo protocols to study endocrine disruption in Drosophila, addressing EDC effects on the fecundity/fertility, developmental timing, and lifespan of the fly. In the last few years, we used these Drosophila life traits to investigate the effects of exposure to 17-&#945;-ethinylestradiol (EE2), bisphenol A (BPA), and bisphenol AF (BPA F). Altogether, these assays covered all Drosophila life stages and made it possible to evaluate endocrine disruption in all hormone-mediated processes. Fecundity/fertility and developmental timing assays were useful to measure the EDC impact on the fly reproductive performance and on developmental stages, respectively. Finally, the lifespan assay involved chronic EDC exposures to adults and measured their survivorship. However, these life traits can also be influenced by several experimental factors that had to be carefully controlled. So, in this work, we suggest a series of procedures we have optimized for the right outcome of these assays. These methods allow scientists to establish endocrine disruption for any EDC or for a mixture of different EDCs in Drosophila, although to identify the endocrine mechanism responsible for the effect, further essays could be needed.

Methods to Test Endocrine Disruption in Drosophila melanogaster

Endocrine disruptor chemicals (EDCs) represent a serious problem for organisms and for natural environments. Drosophila melanogaster represents an ideal model to study EDC effects in vivo. Here, we present methods to investigate endocrine disruption in Drosophila, addressing EDC effects on fecundity, fertility, developmental timing, and lifespan of the fly.

초파리 멜라노가스터에서 내분비 장애를 테스트하는 방법

In recent years there has been growing evidence that all organisms and the environment are exposed to hormone-like chemicals, known as endocrine disruptor ...

발생학

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.

사회 회피의 정량화에 대한 간단한 분석<em&gt; 초파리</em

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

Dyeing Insects for Behavioral Assays: the Mating Behavior of Anesthetized Drosophila

Mating experiments using Drosophila have contributed greatly to the understanding of sexual selection and behavior. Experiments often require simple, easy and cheap methods to distinguish between individuals in a trial. A standard technique for this is CO2 anaesthesia and then labelling or wing clipping each fly. However, this is invasive and has been shown to affect behavior. Other techniques have used coloration to identify flies. This article presents a simple and non-invasive method for labelling Drosophila that allows them to be individually identified within experiments, using food coloring. This method is used in trials where two males compete to mate with a female. Dyeing allowed quick and easy identification. There was, however, some difference in the strength of the coloration across the three species tested. Data is presented showing the dye has a lower impact on mating behavior than CO2 in Drosophila melanogaster. The impact of CO2 anaesthesia is shown to depend on the species of Drosophila, with D. pseudoobscura and D. subobscura showing no impact, whereas D. melanogaster males had reduced mating success. The dye method presented is applicable to a wide range of experimental designs.

Dyeing Insects for Behavioral Assays: the Mating Behavior of Anesthetized Drosophila

This protocol describes a simple, cost effective way to individually identify Drosophila or other insects. Demonstration data investigating mating success across three species of Drosophila show that this method is comparable or better than the use of CO2 anaesthesia.

행동 분석 실험에 대한 염색 곤충 : 마취의 짝짓기 행동<em&gt; 초파리</em

Mating experiments using Drosophila have contributed greatly to the understanding of sexual selection and behavior. Experiments often require simple, easy ...

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.

사회적 공간에 영향을 미치는 조건<em&gt; 초파리</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 ...

A Behavioral Assay for Mechanosensation of MARCM-based Clones in Drosophila melanogaster

Because of the structural and functional homology to the hair cells of the mammalian inner ear, the neurons that innervate the Drosophila external sense organs provide an excellent model system for the study of mechanosensation. This protocol describes a simple touch behavior in fruit flies which can be used to identify mutations that interfere with mechanosensation. The tactile stimulation of a macrochaete bristle on the thorax of flies elicits a grooming reflex from either the first or third leg. Mutations that interfere with mechanotransduction (such as NOMPC), or with other aspects of the reflex arc, can inhibit the grooming response. A traditional screen of adult behaviors would have missed mutants that have essential roles during development. Instead, this protocol combines the touch screen with mosaic analysis with a repressible cell marker (MARCM) to allow for only limited regions of homozygous mutant cells to be generated and marked by the expression of green fluorescent protein (GFP). By testing MARCM clones for abnormal behavioral responses, it is possible to screen a collection of lethal p-element mutations to search for new genes involved in mechanosensation that would have been missed by more traditional methods.

A Behavioral Assay for Mechanosensation of MARCM-based Clones in Drosophila melanogaster

In order to identify novel mutations affecting mechanosensation, we designed an assay that measures the behavioral response to tactile stimulation of fly bristles in mutant clones generated by the MARCM method. The combination of techniques allows for the identification of mechanosensitive mutations that would otherwise be lethal.

의 Mechanosensation에 대한 행동 분석 클론의 MARCM은 기반<em&gt; 초파리</em

Because of the structural and functional homology to the hair cells of the mammalian inner ear, the neurons that innervate the Drosophila external sense ...

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.

사용 보상 추구 행동의 변화를 측정하는 간단한 방법<em&gt; 노랑 초파리</em

We describe a protocol for measuring ethanol self-administration in fruit flies (Drosophila melanogaster) as a proxy for changes in reward states. We ...

Measuring and Altering Mating Drive in Male Drosophila melanogaster

Despite decades of investigation, the neuronal and molecular bases of motivational states remain mysterious. We have recently developed a novel, reductionist, and scalable system for in-depth investigation of motivation using the mating drive of male Drosophila&#160;melanogaster (Drosophila), the methods for which we detail here. The behavioral paradigm centers on the finding that male mating drive decreases alongside fertility over the course of repeated copulations and recovers over ~3 d. In this system, the powerful neurogenetic tools available in the fly converge with the genetic accessibility and putative wiring diagram available for sexual behavior. This convergence allows rapid isolation and interrogation of small neuronal populations with specific motivational functions. Here we detail the design and execution of the satiety assay that is used to measure and alter courtship motivation in the male fly. Using this assay, we also demonstrate that low male mating drive can be overcome by stimulating dopaminergic neurons. The satiety assay is simple, affordable, and robust to influences of genetic background. We expect the satiety assay to generate many new insights into the neurobiology of motivational states.

Measuring and Altering Mating Drive in Male Drosophila melanogaster

This article describes a behavioral assay that uses male mating drive in Drosophila melanogaster to study motivation. Using this method, researchers can utilize advanced fly neurogenetic techniques to uncover the genetic, molecular, and cellular mechanisms that underlie this motivation.

측정과 남성의 변경 결합 드라이브<em&gt; 노랑 초파리</em

Despite decades of investigation, the neuronal and molecular bases of motivational states remain mysterious. We have recently developed a novel, ...

Drosophila Courtship Conditioning As a Measure of Learning and Memory

Many insights into the molecular mechanisms underlying learning and memory have been elucidated through the use of simple behavioral assays in model organisms such as the fruit fly, Drosophila melanogaster. Drosophila is useful for understanding the basic neurobiology underlying cognitive deficits resulting from mutations in genes associated with human cognitive disorders, such as intellectual disability (ID) and autism. This work describes a methodology for testing learning and memory using a classic paradigm in Drosophila known as courtship conditioning. Male flies court females using a distinct pattern of easily recognizable behaviors. Premated females are not receptive to mating and will reject the male&#39;s copulation attempts. In response to this rejection, male flies reduce their courtship behavior. This learned reduction in courtship behavior is measured over time, serving as an indicator of learning and memory. The basic numerical output of this assay is the courtship index (CI), which is defined as the percentage of time that a male spends courting during a 10 min interval. The learning index (LI) is the relative reduction of CI in flies that have been exposed to a premated female compared to na&#239;ve flies with no previous social encounters. For the statistical comparison of LIs between genotypes, a randomization test with bootstrapping is used. To illustrate how the assay can be used to address the role of a gene relating to learning and memory, the pan-neuronal knockdown of Dihydroxyacetone phosphate acyltransferase (Dhap-at) was characterized here. The human ortholog of Dhap-at, glyceronephosphate O-acyltransferase (GNPT), is involved in rhizomelic chondrodysplasia punctata type 2, an autosomal-recessive syndrome characterized by severe ID. Using the courtship conditioning assay, it was determined that Dhap-at is required for long-term memory, but not for short-term memory. This result serves as a basis for further investigation of the underlying molecular mechanisms.

Drosophila Courtship Conditioning As a Measure of Learning and Memory

This protocol describes a Drosophila learning and memory assay called courtship conditioning. This classic assay is based on a reduction of male courtship behavior after sexual rejection by a non-receptive premated female. This natural form of behavioral plasticity can be used to test learning, short-term memory, and long-term memory.

학습과 기억의 척도 로서의 Drosophila Courtship 컨디셔닝

Many insights into the molecular mechanisms underlying learning and memory have been elucidated through the use of simple behavioral assays in model ...

환경에서 신호의 결합 효과에 미치는 영향을 시험하는 방법

요약

더 많은 비디오 탐색

이 비디오의 챕터

분석의 방법 Drosophila 행동

먹이 찾아 돌아다 행동에 대한 단일 비행 분석에

초파리 멜라노가스터에서 내분비 장애를 테스트하는 방법

사회 회피의 정량화에 대한 간단한 분석

행동 분석 실험에 대한 염색 곤충 : 마취의 짝짓기 행동

사회적 공간에 영향을 미치는 조건

의 Mechanosensation에 대한 행동 분석 클론의 MARCM은 기반

사용 보상 추구 행동의 변화를 측정하는 간단한 방법

측정과 남성의 변경 결합 드라이브

학습과 기억의 척도 로서의 Drosophila Courtship 컨디셔닝