フライングストックのブルーミントンショウジョウバエストックセンターに感謝します。 C. Gahr、JT Alkema、およびS. van Hasseltによる加圧空気アッセイの開発に関する初期の試み。ジャスパー・ボスマン、酵母の栽培に関するアドバイス。もともとショウジョウバエの交尾行動の経過観察モニタリングを開発してくれたRezza AzanchiとJoel Levine。 JA Gorterは、Neuroscience Research School BCN / NWO Graduate Programグラントの支援を受けました。この研究は、JC Billeterの科学研究（NWO）（参考文献：821.02.020）のオランダ組織によって一部支持された。

<ol>
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F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sustained+post-mating+response+in+Drosophila+melanogaster+requires+multiple+seminal+fluid+Proteins.">Sustained post-mating response in Drosophila melanogaster requires multiple seminal fluid Proteins.</a> PLoS gen. 3 (12), 2428-2438 (2007).</li><li>Yapici, N., Kim, Y. -. J., Ribeiro, C., Dickson, B. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+receptor+that+mediates+the+post-mating+switch+in+Drosophila+reproductive+behaviour.">A receptor that mediates the post-mating switch in Drosophila reproductive behaviour.</a> Nature. 451 (7174), 33-37 (2008).</li><li>Yang, C. -. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Control+of+the+postmating+behavioral+switch+in+Drosophila+females+by+internal+sensory+neurons.">Control of the postmating behavioral switch in Drosophila females by internal sensory neurons.</a> Neuron. 61 (4), 519-526 (2009).</li><li>H&#228;semeyer, M., Yapici, N., Heberlein, U., Dickson, B. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sensory+neurons+in+the+Drosophila+genital+tract+regulate+female+reproductive+behavior.">Sensory neurons in the Drosophila genital tract regulate female reproductive behavior.</a> Neuron. 61 (4), 511-518 (2009).</li><li>Rez&#225;val, C., Pavlou, H. J., Dornan, A. J., Chan, Y. -. B., Kravitz, E. A., Goodwin, S. 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D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pigment-dispersing+factor+modulates+pheromone+production+in+clock+cells+that+influence+mating+in+Drosophila.">Pigment-dispersing factor modulates pheromone production in clock cells that influence mating in Drosophila.</a> Neuron. 79 (1), 54-68 (2013).</li><li>Imhof, M., Harr, B., Brem, G., Schl&#246;tterer, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multiple+mating+in+wild+Drosophila+melanogaster+revisited+by+microsatellite+analysis.">Multiple mating in wild Drosophila melanogaster revisited by microsatellite analysis.</a> Mol. Ecol. , 915-917 (1998).</li><li>Ochando, M. D., Reyes, A., Ayala, F. 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著者らは、開示する競合する持分はない。

このプロトコルは、交配する夫婦が交配頻度を決定するために使用することが仮定されている環境的合図を連続的に制御しながら、24時間にわたり交配行動を試験するためのアッセイを記載する。培地に酵母も含まれている場合（ 図2B ）、加圧空気システムを介して送達された酵母空気に応答して交配頻度を増加させることが可能である。さらに、寒天、ペプトン、および酢酸?...

生殖行動は、通常、特に女性の場合、男性よりも大きな生殖細胞を産生し、発生する子孫を育てる条件を慎重に選択する必要があるため、エネルギーコストが高い。エネルギーコストのために、再生が栄養状態に関連していることは驚くべきことではない。これは、すべてではないにしても哺乳動物を含むほとんどの動物で、栄養失調により思春期が遅れ、性的欲求が食物制限によって悪影響を受ける可能性がある1に当てはまります。 遺伝的モデル生物Drosophila melanogasterの繁殖はまた、栄養状態の影響を受ける。男性の食品揮発性物質2の存在下でより高いレベルで裁判所、と女性は、酵母の存在、卵の生産と子孫の生存3、4、5のための主要な栄養素で、より性的に受容されています。この進化的に保存された食品への生殖的応答は、遺伝的に扱いやすく時間効率の良い生物において、環境的食物の利用可能性を性的複製と結びつけるメカニズムを研究する機会を提供する。実際、 D. melanogasterの研究は、インスリン経路が食物と交配行動との関係の重要な調節因子であることを示唆している6 。それはまた、それ自体を相手の行為が女性の食品嗜好、ならびに関連する化学感覚ニューロン7、8、9を変えることが示されています。 D. melanogasterの食餌の合図は生殖行動に影響することは明らかである。これらの影響は主に女性、特に既に交配している女性に影響するようである5 。しかしながら、環境条件のこれらの急性効果を試験するために、雌交配行動に古典的に用いられるアッセイは、交配エピソードの間に長い間中断があるため、あまり適切ではない。古典的なrematingアッセイでは、処女の女性は最初に男性と仲良くなり、すぐに隔離され、24〜48時間後に新しい男性が呈示されます。この古典的なアッセイは、女性の行動と雌応答12、13、14、15、16、17、18を変更男性精液の成分を同定するために成功裏に使用されてきました。したがって、ここで示された連続交配アッセイは、生殖行動に対する環境条件の急性効果を研究するために使用され得る古典的な交配アッセイに加えられるものである。 ここで説明されている交尾行動についての連続アッセイを用いて、以前に酵母に曝露されたハエのペアが、24時間の観察期間5、19、20を超えるeveral回、21、ハエが食品に曝されていない間は一度だけ5 Remateのであろう。この知見は、女性が（参照10、11に概説されている）最初の交配後、数日間Remateのないことを示すキイロショウジョウバエの文学の大部分の光の中で不可解なことができます。しかし、この相違は、新しい交配機会が提供される前に、1〜数日間、女性が単離されるアッセイ条件によって容易に説明することができる。ペアがこの1時間の観察期間に交配しなければ、女性は受容性ではないと特徴付けられる。さらに、野生の捕獲されたハエからのデータが、女性が貯蔵器官に4〜6匹の男性の精子を含むことを示すとすれば、高い交配頻度は驚くべきではない。したがって、女性は自然に数回22、23を Remateのことdicating。 ここでは、この連続交配アッセイを使用して、ハエがどのように集まり、環境条件に関する情報を組み合わせて交配頻度を調節するかを実証する。このアッセイにより、遺伝学的研究のために比較的多数の交配対を試験し、揮発性及び非揮発性の環境手がかりの影響を試験することができる。アッセイは典型的には24時間実行されるが、48時間まで延長することができ、明暗（LD）サイクルなどの循環環境試験を行うことができる。我々は、食品基質中の不揮発性酵母栄養素の利用可能性と相まって、加圧空気システム内の酵母培養からの揮発性手がかりの影響を試験することにより、このアッセイを実証する。 加圧空気システムは、揮発性の手がかりを、食物基質および試験対（その交配挙動がモニターされる）。酵母が交配に影響を及ぼす特異性をさらに決定するために、本発明者らは、酵母の主要な揮発性化合物、すなわち酢酸24を、食品基質中の酵母のアミ​​ノ酸含量と組み合わせて、ペプトン（アミノ動物性タンパク質の酵素消化に由来する酸）。これらの実験は、 D. melanogasterの交配行動に対する環境上の合図の効果がこのアッセイでどのように試験され得るかを示す。

<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

個人の性的欲求は、遺伝子型、経験および環境条件の影響を受ける。性的行動を調節するためにこれらの因子がどのように相互作用するかは、依然として十分に理解されていない。 ショウジョウバエのメラノガスターでは 、食物の利用可能性などの環境上の手がかりが、性行動を調節するメカニズムを調べるための扱いやすいシステムを提供する交配活動に影響を与えます。 D. melanogasterでは、化学感覚性の味覚システムと嗅覚系を介して、環境の合図がしばしば感知されます。ここでは、交尾行動に対する環境化学的合図の効果を試験する方法を提示する。アッセイは、食物培地と交配するカップルを含む小さな交配アリーナからなる。各カップルの交配頻度を24時間連続してモニターする。ここでは、加圧空気システムを介した外部供給源からの環境化合物の試験、および対合アリーナ内の環境成分の直接操作に対するこのアッセイの適用性を示します。あなたは非常に揮発性の化合物の効果を試験するのに特に有用である一方、交配アリーナ内の成分を直接操作することは、化合物の存在を確かめるために有益であり得る。このアッセイは、交配行動および産卵ならびに他の男性および女性の生殖行動に及ぼす遺伝的および環境的合図の影響に関する質問に答えるために適合させることができる。

この連続アッセイを使用して、交配行動および特異的な交配頻度を、実験的環境条件下で決定することができる。環境条件をコントロールするために、ステンレス製のキッチンキャビネットを、独自の光源と拡散機能を備えたテストエリアに変えました。これにより、光が豊富になり、対合アリーナの上部からグレアが最小限に抑えられました（ 図1A ） ...

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

我々はショウジョウバエDrosophila melanogasterの交尾行動に影響を与える環境および遺伝的合図を分析するためのアッセイを実証する。

環境内の交信が交配行動に及ぼす影響を調べる方法<em&gt;ショウジョウバエ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 ビュー.  University of Groningen. 我々はショウジョウバエDrosophila melanogasterの交尾行動に影響を与える環境および遺伝的合図を分析するためのアッセイを実証する。 

ビデオ: 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.

アッセイのメソッドショウジョウバエ行動

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

神経科学

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.

での採餌行動のためのシングル·フライアッセイ<em&gt;ショウジョウバエ</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.

ショウジョウバエメラメラノガスターにおける内分泌破壊を試験する方法

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

発生生物学

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.

社会回避での定量化のための簡単​​なアッセイ<em&gt;キイロショウジョウバエ</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.

行動アッセイの染色昆虫：麻酔の配偶行動<em&gt;ショウジョウバエ</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.

で社会空間に影響を与える条件<em&gt;キイロショウジョウバエ</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.

でMARCMベースのクローンのMechanosensationための行動アッセイ<em&gt;キイロショウジョウバエ</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.

使用して報酬を求める行動の変化を測定するための簡単な方法<em&gt;キイロショウジョウバエ</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.

測定とマレの変更交尾ドライブ<em&gt;キイロショウジョウバエ</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.

ショウジョウバエの訓練と記憶の尺度としてのコンディショニング

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

環境内の交信が交配行動に及ぼす影響を調べる方法

要約

さらに動画を探す

この動画の章

アッセイのメソッドショウジョウバエ行動

での採餌行動のためのシングル·フライアッセイ

ショウジョウバエメラメラノガスターにおける内分泌破壊を試験する方法

社会回避での定量化のための簡単なアッセイ

行動アッセイの染色昆虫：麻酔の配偶行動

で社会空間に影響を与える条件

でMARCMベースのクローンのMechanosensationための行動アッセイ

使用して報酬を求める行動の変化を測定するための簡単な方法

測定とマレの変更交尾ドライブ

ショウジョウバエの訓練と記憶の尺度としてのコンディショニング

環境内の交信が交配行動に及ぼす影響を調べる方法

要約

さらに動画を探す

この動画の章

アッセイのメソッドショウジョウバエ行動

での採餌行動のためのシングル·フライアッセイ

ショウジョウバエメラメラノガスターにおける内分泌破壊を試験する方法

社会回避での定量化のための簡単​​なアッセイ

行動アッセイの染色昆虫：麻酔の配偶行動

で社会空間に影響を与える条件

でMARCMベースのクローンのMechanosensationための行動アッセイ

使用して報酬を求める行動の変化を測定するための簡単な方法

測定とマレの変更交尾ドライブ

ショウジョウバエの訓練と記憶の尺度としてのコンディショニング

社会回避での定量化のための簡単なアッセイ