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

社内で作成1.機器および試薬（他のためのマテリアルのリストを参照してください） <ol><li>フライ作業を実行するために、ショウジョウバエ冷たい麻酔装置を準備します。 <ol><li>小さなプラスチックの箱（10.2センチ幅12.7センチ）、ピペットチップボックスの一般的に、トップカバー（3.8センチメートル深い10.2センチ幅12.7センチ）を覆うように多孔質ポリエチレンシートをカット。 </li><li>砕いた氷の箱を埋める、多孔性ポリエチレンシートでカバーしています。 </li></ol></li><li> T-迷路装置を準備します。以前に詳細28-31に記載された装置を適応させる。 </li><li> 32は説明したように、フライアスピレーターを準備します。 </li><li>ストレスなくテストバイアルの内外ハエを変位させる走光性応答装置を準備します。 <ol><li> 29,33に記載されているように、向流装置を使用してください。 </li></ol></li></ol> NOTE：28によって記載されるようにT字型迷路はまた、他の目的に使用することができる。 <ol start="5"><li> HOMを確認してくださいogeneous照明条件は：白ベンチカバーで覆われたベンチに実験を行い、ホワイトボードの前に。 注：これらの実験のために、使用されるフライ株はショウジョウバエた野生型をキイロ ：カントン-S、オレゴン州とサマルカンドは、私たちの研究室のストックから34を飛ぶ。エルウッドハエはロングアイランド、ニューヨーク、米国で、ハンチントンのエルウッド周辺に秋2011年に収集した。 </li></ol> 2.実験前にハエの準備実験を行う前に1〜2日： <ol><li> 12時間の明/暗サイクルで25℃における標準コーンミール寒天 - 糖蜜 - 酵母媒体上のハエを維持します。 </li><li>実験を行うことに1〜2日前に風邪麻酔下のレスポンダハエを収集<ol><li>彼らは漏斗を用いて、50ミリリットルファルコンチューブに提起されている中でボトルからハエを転送します。 </li><li>に、氷のレベルの下のハエでファルコンチューブを置きます絶縁されたアイスバケット。 </li><li>冷却するで-4℃、砕いた氷と冷たい麻酔装置を配置します。 </li><li>ハエは寒さ昏睡を入力するための5分待ちます。 </li><li>冷たい麻酔装置の多孔質ポリエチレンシート上にハエを転送します。 </li><li>冷たい麻酔装置上​​のレスポンダハエを収集します。 RTで、実体顕微鏡下で、女性のハエから3-7日歳の男性を分離し、40のグループでこれらのハエを維持する。 </li></ol></li><li>実験を行うことに1〜2日前にエミッタハエを収集します。 <ol><li>静かにカントン-S混在セックスはその数をコントロールし、口の吸引器を使用して、彼らのボトルから飛ぶ集める。 </li><li>彼らはエミッタであるように、60〜100のグループでこれらのハエを維持します。 </li><li>データ点は、必要に応じて発光体のように多くのサンプルを準備します。 </li></ol></li><li>冷たい麻酔のいずれかの交絡影響を最小化するために、応答者およびエミッタの収集後の回復の少なくとも夜を許可する行動に、口アスピレーターコレクション。 </li><li>少なくとも1日実験前に、新鮮な空気に置き換えられるように袋に入れて古くなったプラスチック製の香りの空気を可能にするために、テスト用バイアルの袋を開きます。 </li></ol>実験を行う前に、2時間。 <ol start="6"><li>でも光で、実験が行われる室内の温度は約23-25​​℃であることを確認し、30％の上記の湿度。 </li><li>これらのハエが飢えていないことを確実にするために生鮮食品バイアルに応答者およびエミッタショウジョウバエを転送します。 </li><li>ショウジョウバエは、実験が実行される上のベンチに、2時間の環境に適応してみましょう。 </li></ol> 3.ソーシャル回避実験を行うこと<ol><li> Zeitgeber時間ZT5とZT9間午後3〜4時間の範囲で、一日の同じ時刻に、それぞれの実験を行います。 </li><li>ドキドキパッド上でT-迷路を置き、およびtので、ネジクリップを締め彼はエレベーター安定している。 </li><li>新鮮な試験バイアルにレスポンダハエを転送します。 T-迷路の上部に、レスポンダハエを含むこのバイアルをスナップ。 </li><li> T-迷路装置を傾斜し、ショウジョウバエがエレベーターに落ちるようにドキドキマットの上をタップします。 </li><li>ハエは、T-迷路の上部と下部の間にあるので、ダウンエレベーターの上部に移動しますが、選択点以上のエレベーターを維持するために注意を払う - 逃げるのハエを防ぐために。 </li><li> DSO（以前にハエによって占有されたDSOのないバイアルを生成するために、第5節を参照）のバイアルを取得します。 <ol><li>応答者のハエは、この新しい環境に適応しているが、試験バイアルにエミッタハエを配置。 </li><li>ミバエが逃げないように、バイアルを閉じるために綿の部分を置きます。 </li><li> 15秒間ボルテックスを5秒間ボルテックスからバイアルを取り外し、次のように機械的にミニ渦にこのバイアルを攪拌する。 55秒の合計3回繰り返します。 </li><li>レム食品バイアルに転送することによって - - エミッタOVE新鮮なバイアルから飛ぶとすぐにT-迷路装置の両側のいずれかにこのDSO満たされたバイアルを置く。 </li><li>それぞれの新しい目でDSO埋めバイアルの配置の側面を交互に。 </li></ol></li><li> T-迷路の反対側に新鮮な試験バイアルを置きます。 </li><li>ショウジョウバエは、新鮮な試験バイアルとDSOで試験バイアルの間で選択することができるように、完全にダウンしてエレベーターを持参し、タイマーを開始。 </li><li>特に断りのない限り、ハエは、1分間、新鮮なバイアルとDSOバイアルのいずれかを選択しましょう​​。 </li><li> 1分後、新鮮なバイアル、DSOバイアル、およびエレベーターの中で立ち往生ものでハエを分離するためにエレベーターを上に移動。 </li><li>各バイアルにハエの数をカウントし、エレベーターで。 </li><li>各遺伝子型/条件のためにこれを繰り返します。 </li></ol> 4.ソーシャル回避データの分析<ol><li>各ジュノのための各バイアルにミバエの数を数えYPE、およびスプレッドシートにデータを転送。 </li><li>表計算プログラムを使用して、DSOのないバイアル中のハエの数からDSOバイアルにハエの数を減算し、その後、ハエの総数で割ることによって、各遺伝子型のための性能指数（PI）を計算する。 </li><li>結果の分析のための統計解析ソフトを用いてPIは「手段の比較。 <ol><li>理想的には、各実験のための2-3内部反復、および3つの独立した繰り返しの準備、別のボトルで、異なる日に行わまたは30を横切る。 </li><li>データが正規分布に従うように、ガウス分布の統計的検定を使用してください。 </li><li> 1効果をテストするとき（ すなわち 、密度- 図2）、コントロールにそれぞれの結果を比較するために、（列ごと）の多重比較のための試験後、または単純なt検定で、一方向ANOVAを使用しています。 </li><li>グループ実験が行われた場合、条件および性別（空のバイアルの両方をテストし、強調Dの未強調したハエ。ジェンダー影響- 図1）は 、双方向ANOVAを使用して。 </li></ol></li></ol> 5.以前はストレスのないハエによって占有バイアルの生成 - その代わりに3.5から3.6の実行<ol><li>コントロールとして、強調されていないハエが占有するバイアルを準備する走光性を使用してください。 </li><li>試験バイアルにハエを転送し、反対の場所で新鮮な空のバイアルで、向流装置にはめ込みます。 </li><li> 15 W蛍光クール白色光の前で直接遠位バイアルで水平に装置を置き、そして黒の布でカバーしています。 </li><li>ハエが優しく新鮮なバイアルに移動しましょう​​。これには数分かかることがあります。ハエのほとんどは遠位バイアル中の光に向かって歩いていると、1分間のタイマーを起動します。 </li><li>静かに、黒い布を削除する光の位置を切り替えて、黒い布で回復し、ハエが試験バイアルを残すことができます。 </li><li>その時、私3.2から3.5のように選択肢実験をnitiate。 3.6と3.7をスキップします。 </li><li>他の側からの試験バイアルを空いたハエを分離するために、装置のスライダを移動します。 </li><li> 3.7のように、選択肢の実験のためにすぐにT-迷路に空いた試験バイアルを使用してください。 </li></ol>

<ol>
	<li>Fry, S. N., Rohrseitz, N., Straw, A. D., Dickinson, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=TrackFly:+virtual+reality+for+a+behavioral+system+analysis+in+free-flying+fruit+flies.">TrackFly: virtual reality for a behavioral system analysis in free-flying fruit flies.</a> J Neurosci Methods. 171, 110-117 (2008).</li><li>Slawson, J. B., Kim, E. Z., Griffith, L. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-resolution+video+tracking+of+locomotion+in+adult+Drosophila+melanogaster.">High-resolution video tracking of locomotion in adult Drosophila melanogaster.</a> J Vis Exp. 24, 1096 (2009).</li><li>Dankert, H., Wang, L., Hoopfer, E. D., Anderson, D. J., Perona, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Automated+monitoring+and+analysis+of+social+behavior+in+Drosophila.">Automated monitoring and analysis of social behavior in Drosophila.</a> Nat Methods. 6, 297-303 (2009).</li><li>Branson, K., Robie, A. A., Bender, J., Perona, P., Dickinson, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-throughput+ethomics+in+large+groups+of+Drosophila.">High-throughput ethomics in large groups of Drosophila.</a> Nat Methods. 6, 451-457 (2009).</li><li>Simon, J. C., Dickinson, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+chamber+for+studying+the+behavior+of+Drosophila.">A new chamber for studying the behavior of Drosophila.</a> PLoS One. 5, 8793 (2010).</li><li>Wang, L., Dankert, H., Perona, P., Anderson, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inaugural+Article:+A+common+genetic+target+for+environmental+and+heritable+influences+on+aggressiveness.">Inaugural Article: A common genetic target for environmental and heritable influences on aggressiveness.</a> in Drosophila. Proceedings of the National Academy of Sciences. 105, 5657-5663 (2008).</li><li>Schneider, J., Dickinson, M. H., Levine, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Social+structures+depend+on+innate+determinants+and+chemosensory+processing+in+Drosophila.">Social structures depend on innate determinants and chemosensory processing in Drosophila.</a> Proceedings of the National Academy of Sciences. 2, 17174-17179 (2012).</li><li>Miyamoto, T., Amrein, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Suppression+of+male+courtship+by+a+Drosophila+pheromone+receptor.">Suppression of male courtship by a Drosophila pheromone receptor.</a> Nat Neurosci. 11, 874-876 (2008).</li><li>Villella, A., Hall, J. C., Jeffrey, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chapter+3+Neurogenetics+of+Courtship+and+Mating+in+Drosophila.+.">Chapter 3 Neurogenetics of Courtship and Mating in Drosophila. .</a> Advances in Genetics. 62, 67-184 (2008).</li><li>Ejima, A., Griffith, L. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Courtship+Initiation+Is+Stimulated+by+Acoustic+Signals+in+Drosophila+melanogaster.">Courtship Initiation Is Stimulated by Acoustic Signals in Drosophila melanogaster.</a> PLoS ONE. 3, 3246 (2008).</li><li>Mery, F., 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=Public+Versus+Personal+Information+for+Mate+Copying+in+an+Invertebrate.">Public Versus Personal Information for Mate Copying in an Invertebrate.</a> Current Biology. 19, 730-734 (2009).</li><li>Montell, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+taste+of+the+Drosophila+gustatory+receptors.">A taste of the Drosophila gustatory receptors.</a> Curr. Opin. Neurobiol. 19, 345-353 (2009).</li><li>Billeter, J. -. C., Atallah, J., Krupp, J. J., Millar, J. G., Levine, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Specialized+cells+tag+sexual+and+species+identity+in+Drosophila+melanogaster.">Specialized cells tag sexual and species identity in Drosophila melanogaster.</a> Nature. 461, 987-991 (2009).</li><li>Krupp, J. J., 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=Social+experience+modifies+pheromone+expression+and+mating+behavior+in+male+Drosophila+melanogaster.">Social experience modifies pheromone expression and mating behavior in male Drosophila melanogaster.</a> Curr Biol. 18, 1373-1383 (2008).</li><li>Kent, C., Azanchi, R., Smith, B., Formosa, A., Levine, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Social+context+influences+chemical+communication+in+D.+melanogaster+males.">Social context influences chemical communication in D. melanogaster males.</a> Curr Biol. 18, 1384-1389 (2008).</li><li>Levine, J. D., Funes, P., Dowse, H. B., Hall, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Resetting+the+Circadian+Clock+by+Social+Experience+in+Drosophila+melanogaster.">Resetting the Circadian Clock by Social Experience in Drosophila melanogaster.</a> Science. 298, 2010-2012 (2002).</li><li>Ganguly-Fitzgerald, I., Donlea, J., Shaw, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Waking+Experience+Affects+Sleep+Need+in+Drosophila.">Waking Experience Affects Sleep Need in Drosophila.</a> Science. 313, 1775-1781 (2006).</li><li>Billeter, J. -. C., Levine, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Who+is+he+and+what+is+he+to+you?+Recognition+in+Drosophila+melanogaster.">Who is he and what is he to you? Recognition in Drosophila melanogaster.</a> Curr. Opin. Neurobiol. 23, 17-23 (2013).</li><li>Suh, G. S., 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=A+single+population+of+olfactory+sensory+neurons+mediates+an+innate+avoidance+behaviour+in+Drosophila.">A single population of olfactory sensory neurons mediates an innate avoidance behaviour in Drosophila.</a> Nature. 431, 854-859 (2004).</li><li>Bonini, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Tribute+to+Seymour+Benzer+1921-2007.">A Tribute to Seymour Benzer 1921-2007.</a> 180, 1265-1273 (2008).</li><li>Castermans, D., 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=The+neurobeachin+gene+is+disrupted+by+a+translocation+in+a+patient+with+idiopathic+autism.">The neurobeachin gene is disrupted by a translocation in a patient with idiopathic autism.</a> Journal of Medical Genetics. 40, 352-356 (2003).</li><li>Medrihan, L., 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=Neurobeachin,+a+protein+implicated+in+membrane+protein+traffic+and+autism,+is+required+for+the+formation+and+functioning+of+central+synapses.">Neurobeachin, a protein implicated in membrane protein traffic and autism, is required for the formation and functioning of central synapses.</a> J Physiol. 587, 5095-5106 (2009).</li><li>Simon, A. F., 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=A+simple+assay+to+study+social+behavior+in+Drosophila:+measurement+of+social+space+within+a+group.">A simple assay to study social behavior in Drosophila: measurement of social space within a group.</a> Genes Brain Behav. 11, 243-252 (2012).</li><li>Venkatesh, T., 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=.">.</a> Cold Spring Harbor Meeting: From Molecules to Circuit Behavior. , (2013).</li><li>Hamilton, P. J., 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=De+novo+mutation+in+the+dopamine+transporter+gene+associates+dopamine+dysfunction+with+autism+spectrum+disorder.">De novo mutation in the dopamine transporter gene associates dopamine dysfunction with autism spectrum disorder.</a> Mol Psychiatry. 18, 1315-1323 (2013).</li><li>Gadow, K. D., 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=Association+of+dopamine+gene+variants,+emotion+dysregulation+and+ADHD+in+autism+spectrum+disorder.">Association of dopamine gene variants, emotion dysregulation and ADHD in autism spectrum disorder.</a> Research in Developmental Disabilities. 35, 1658-1665 (2014).</li><li>Fernandez, R. W., Akinleye, A. A., Nurilov, M., Rouzyi, Z., Simon, A. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> , (2014).</li><li>Ali, Y. O., Escala, W., Ruan, K., Zhai, R. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assaying+Locomotor,+Learning,+and+Memory+Deficits+in+Drosophila+Models+of+Neurodegeneration.">Assaying Locomotor, Learning, and Memory Deficits in Drosophila Models of Neurodegeneration.</a> J Vis Exp. , 2504 (2011).</li><li>Connolly, J. B., Tully, T., Roberts, D. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Drosophila: A Practical Approach. 1, 265-317 (1998).</li><li>Tully, T., Quinn, W. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Classical-conditioning+and+retention+in+normal+and+mutant+Drosophila+melanogaster.">Classical-conditioning and retention in normal and mutant Drosophila melanogaster.</a> J Comp Physiol [A]. 157, 263-277 (1985).</li><li>Krashes, M. J., Waddell, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drosophila+Aversive+Olfactory+Conditioning.">Drosophila Aversive Olfactory Conditioning.</a> Cold Spring Harbor Protocols. 2011, (2011).</li><li>Ejima, A., Griffith, L. C., Zhang, B., Freeman, M. R., Waddell, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ch.+30.">Ch. 30.</a> Drosophila Neurobiology, A Laboratory Manual. , 475-481 (2010).</li><li>Benzer, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+mutants+of+Drosophila+melanogaster+isolated+by+countercurrent+distribution.">Behavioral mutants of Drosophila melanogaster isolated by countercurrent distribution.</a> PNAS. 58, 1112-1119 (1967).</li><li>Simon, A. F., Shih, C., Mack, A., Benzer, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Steroid+control+of+longevity+in+Drosophila+melanogaster.">Steroid control of longevity in Drosophila melanogaster.</a> Science. 299, 1407-1410 (2003).</li><li>Vaux, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Research+methods:+Know+when+your+numbers+are+significant.">Research methods: Know when your numbers are significant.</a> Nature. 492, 180-181 (2012).</li><li>Stowers, L., Logan, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sexual+dimorphism+in+olfactory+signaling.">Sexual dimorphism in olfactory signaling.</a> Curr. Opin. Neurobiol. 20, 770-775 (2010).</li><li>Simon, A. F., Liang, D. T., Krantz, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+decline+in+behavioral+performance+of+Drosophila+melanogaster+with+age.">Differential decline in behavioral performance of Drosophila melanogaster with age.</a> Mechanisms of Ageing and Development. , 127-647 (2006).</li></ol>

The authors declare that they have no competing financial interests.

このプロトコルは、社会的な回避アッセイのための詳細な手順を説明します。カントン-Sのみが飛んでいるバイアルを避けることができます以前に機械的に強調されてきた、と応答者のその性別と数は、その社会的な回避性能には影響しません。しかし、応答者の遺伝的背景に大きな影響を有する。 以下では正常にこの実験を実施するためのいくつかの重要なステップで...

この方法の目的は、容易に求愛と侵略から独立したキイロショウジョウバエにおける単純な社会的行動の新たな側面を定量化することである。 社会的相互作用は、社会の中の個人の適切な発達と健康に重要であるだけでなく、全体としての社会集団の機能性。これらの相互作用の複雑性の高い大規模なサンプルサイズおよび社会的行動の遺伝的及び神経塩基はまだよく理解されて動作の簡略化を可能にするシステムが必要となる。 キイロショウジョウバエの遺伝子を同定するために使用できる強力な遺伝モデルである社会的相互作用の神経基盤。実際、D.キイロは、複雑な社会的行動のレパートリーを持ち、社会のいくつかの直接測定は、すでに1-7行われている。しかしながら、これらの努力のほとんどは、aggresのような比較的複雑な社会的行動に焦点を当ててきたSIVEの相互作用3,6、求愛3,8-12のさまざまな側面、そしてどのように社会的な経験は、学習、または概日リズム13-17のような他の行動に影響を与えます。また、これらのアッセイの多くは、結果として生じるデータの存在量を分析するためにビデオトラッキングとコンピュータソフトウェアを使用して、ハエの群の複雑な相互作用のパターンを分析に依存する。このような分析は非常に貴重であり、そのようなグループ7のフライフライ相互作用の動的などの重要な新しい洞察をもたらす。一つの制限は、しかしながら、一般社会にこれらのアッセイの到達不能などの認識の基礎となるメカニズムの限られた知識である。つまり、一人の個人と他者によるその認識による信号の放射の基礎はまだよく理解されている18。 D.：これとは対照的に、個人が離れてストレスをハエによって放出された信号から移動するシンプルな行動、社会的な回避を呈するもハエメランストレス匂いやDSO 19 ogaster。ハイスループットアッセイでは、この動作は、他のハエ、または社会的回避19によって放射されるストレスシグナルの回避のように定量化することができる。ハエをT迷路装置に入れ、DSOを含むバイアルを回避する選択肢を与えられている。このアッセイを用いて、CO 2は、DSOの成分、および2 19を切開したCOに応答するのに必要な神経回路の一部であることが示された。 ここに提示され、社会的回避アッセイは、概念的に、複雑な振る舞い20を解剖するために、研究者の世代を有効にシーモア·ベンザーの研究室で開発されたシンプルな行動アッセイに似ています。社会的回避の分析は、社会的行動のより広範な研究を可能にする、T迷路アッセイを用いてコスト効率よく行うことができる。例えば、このアッセイを使用して、我々は最近実証自閉症の異なる遺伝的リスクは、社会的なB内の対照的な効果を有することehaviorアッセイ。自閉症の候補遺伝子についての変異体- 21,22 neurobeachin - （他の場所で23を参照）の両方の社会的空間に存在する欠陥および社会的な回避24。異常なドーパミン作動性シグナル伝達はまた、ヒト25,26における自閉症の病因に役割を果たすことが提案されている。 neurobeachinで得られた結果とは対照的に、社会的空間を直接VMAT 27のこれらのレベルと相関したが、社会的回避のパフォーマンスは、ドーパミン作動性細胞ショウジョウバエ胞性モノアミン輸送体（VMAT）の増加または減少したレベルによって影響されなかったことを見出した。 neurobeachinとVMATで得られた対照的な結果は、反社会的行動の様々な形態を識別するため、異なる基礎となる神経回路網が他の応答を調節する可能性を強調する。

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Stereo Zoom Microscope&#160;</td>
			<td>Nikon&#160;&#160;</td>
			<td>SMZ-645</td>
			<td>Any other standard scope for fly handling would work</td>
		</tr>
		<tr>
			<td>Small paint brushes&#160;</td>
			<td></td>
			<td></td>
			<td>for pushing flies</td>
		</tr>
		<tr>
			<td>Porous Polyethylene, 12&#34; x 12&#34; Sheet</td>
			<td>Flystuff - Genesee</td>
			<td>46-100</td>
			<td>http://www.flystuff.com/ProductInfo.php?productID=46-100 
			Porous Plastic sheet for the cold anesthesia box</td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Mini-Alarm Timer/Stopwatch</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>&#160;Sharpie pens</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Adhesive Tape</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Mini vortex</td>
			<td>Fisher</td>
			<td>14-955-151&#160;</td>
			<td>http://www.fishersci.com/ecomm/servlet/itemdetail 
			For mechanical agitation of the flies - any vortex would work.</td>
		</tr>
		<tr>
			<td>Corning Life Sciences DL No.:352017, Falcon test tube; round bottom; disposable; no closure, 14mL;&#160; 17 x 100mm</td>
			<td>Fisher</td>
			<td>14-959-8 &#160;&#160;</td>
			<td>http://www.fishersci.com/ecomm/servlet/itemdetail?storeId=10652&#38;langId=-1&#38;catalogId=29104&#38;productId=2771811&#38;distype=0&#38;highlightProductsItemsFlag=Y&#38;fromSearch=1&#38;searchType=PROD&#38;hasPromo=0 
			These snap in place in the in-house made T-maze and counter-current apparatus (see text)</td>
		</tr>
		<tr>
			<td>cotton balls</td>
			<td></td>
			<td></td>
			<td>to close the vials after the experiment.</td>
		</tr>
		<tr>
			<td>trifold board and white bench cover</td>
			<td></td>
			<td></td>
			<td>to provide a white background, and a homogeneous light.</td>
		</tr>
		<tr>
			<td>white bench cover</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>pounding pad</td>
			<td></td>
			<td></td>
			<td>any mouse pad works.</td>
		</tr>
		<tr>
			<td>large black cloth</td>
			<td></td>
			<td></td>
			<td>to cover the counter-current apparatus in phototaxis response.</td>
		</tr>
		<tr>
			<td>cool-white light&#160;</td>
			<td>Home Depot</td>
			<td>1000516563</td>
			<td>http://www.homedepot.ca/product/illume-26-fluorescent-plug-in-linear/911423 
			any similar linear light with fluorescent light bulb cool-white at 13-15W would work</td>
		</tr>
	</tbody>
</table>

straightforward assay for quantification of social avoidance in drosophila melanogaster

Drosophila melanogaster is an emerging model to study different aspects of social interactions. For example, flies avoid areas previously occupied by stressed conspecifics due to an odorant released during stress known as the Drosophila stress odorant (dSO). Through the use of the T-maze apparatus, one can quantify the avoidance of the dSO by responder flies in a very affordable and robust assay. Conditions necessary to obtain a strong performance are presented here. A stressful experience is necessary for the flies to emit dSO, as well as enough emitter flies to cause a robust avoidance response to the presence of dSO. Genetic background, but not their group size, strongly altered the avoidance of the dSO by the responder flies. Canton-S and Elwood display a higher performance in avoiding the dSO than Oregon and Samarkand strains. This behavioral assay will allow identification of mechanisms underlying this social behavior, and the assessment of the influence of genes and environmental conditions on both emission and avoidance of the dSO. Such an assay can be included in batteries of simple diagnostic tests used to identify social deficiencies of mutants or environmental conditions of interest.

ソーシャル回避アッセイは、他のハエによって放出されるストレスシグナル（DSO）を認識するキイロショウジョウバエの能力を定量化する堅牢な試験であるため、社会的相互作用の一つの側面を評価する。このアッセイは、一般に、2つの異なるオプションの選択にハエを提示するT字型迷路、として知られている種々の行動アッセイで使用される装置を用いて行われる-左または右19...

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

Straightforward Assay for Quantification of Social Avoidance in Drosophila melanogaster

Here, we present a protocol to quantify the avoidance of stressed individuals. This paradigm is powerful yet user-friendly and can be used to assess the influence of genes and environment on one kind of social interaction in Drosophila melanogaster.

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 ...

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

Research

JoVE Journal

Neuroscience

Straightforward Assay for Quantification of Social Avoidance in Drosophila melanogaster

11.1K ビュー.  Yale University. Here, we present a protocol to quantify the avoidance of stressed individuals. This paradigm is powerful yet user-friendly and can be used to assess the influence of genes and environment on one kind of social interaction in Drosophila melanogaster.

ビデオ: Straightforward Assay for Quantification of Social Avoidance in Drosophila melanogaster

Paired Nanoinjection and Electrophysiology Assay to Screen for Bioactivity of Compounds using the Drosophila melanogaster Giant Fiber System

Screening compounds for in vivo activity can be used as a first step to identify candidates that may be developed into pharmacological agents1,2. We developed a novel nanoinjection/electrophysiology assay that allows the detection of bioactive modulatory effects of compounds on the function of a neuronal circuit that mediates the escape response in Drosophila melanogaster3,4. Our in vivo assay, which uses the Drosophila Giant Fiber System (GFS, Figure 1) allows screening of different types of compounds, such as small molecules or peptides, and requires only minimal quantities to elicit an effect. In addition, the Drosophila GFS offers a large variety of potential molecular targets on neurons or muscles. The Giant Fibers (GFs) synapse electrically (Gap Junctions) as well as chemically (cholinergic) onto a Peripheral Synapsing Interneuron (PSI) and the Tergo Trochanteral Muscle neuron (TTMn)5. The PSI to DLMn (Dorsal Longitudinal Muscle neuron) connection is dependent on D&alpha;7 nicotinic acetylcholine receptors (nAChRs)6. Finally, the neuromuscular junctions (NMJ) of the TTMn and the DLMn with the jump (TTM) and flight muscles (DLM) are glutamatergic7-12. Here, we demonstrate how to inject nanoliter quantities of a compound, while obtaining electrophysiological intracellular recordings from the Giant Fiber System13 and how to monitor the effects of the compound on the function of this circuit. We show specificity of the assay with methyllycaconitine citrate (MLA), a nAChR antagonist, which disrupts the PSI to DLMn connection but not the GF to TTMn connection or the function of the NMJ at the jump or flight muscles.
Before beginning this video it is critical that you carefully watch and become familiar with the JoVE video titled "Electrophysiological Recordings from the Giant Fiber Pathway of D. melanogaster " from Augustin et al7, as the video presented here is intended as an expansion to this existing technique. Here we use the electrophysiological recordings method and focus in detail only on the addition of the paired nanoinjections and monitoring technique.

Paired Nanoinjection and Electrophysiology Assay to Screen for Bioactivity of Compounds using the Drosophila melanogaster Giant Fiber System

A rapid in vivo assay to test for neuromodulatory compounds using the Giant Fiber System (GFS) of Drosophila melanogaster is described. Nanoinjections in the head of the animal along with electrophysiological recordings of the GFS can reveal bioactivity of compounds on neurons or muscles.

用いて、化合物の生理活性をスクリーニングするためのペアNanoinjectionと電気生理学的アッセイキイロショウジョウバエジャイアントファイバシステム

Screening compounds for in vivo activity can be used as a first step to identify candidates that may be developed into pharmacological agents1,2. We ...

神経科学

Optogenetic Stimulation of Escape Behavior in Drosophila melanogaster

A growing number of genetically encoded tools are becoming available that allow non-invasive manipulation of the neural activity of specific neurons in Drosophila melanogaster1. Chief among these are optogenetic tools, which enable the activation or silencing of specific neurons in the intact and freely moving animal using bright light. Channelrhodopsin (ChR2) is a light-activated cation channel that, when activated by blue light, causes depolarization of neurons that express it. ChR2 has been effective for identifying neurons critical for specific behaviors, such as CO2 avoidance, proboscis extension and giant-fiber mediated startle response2-4. However, as the intense light sources used to stimulate ChR2 also stimulate photoreceptors, these optogenetic techniques have not previously been used in the visual system. Here, we combine an optogenetic approach with a mutation that impairs phototransduction to demonstrate that activation of a cluster of loom-sensitive neurons in the fly's optic lobe, Foma-1 neurons, can drive an escape behavior used to avoid collision. We used a null allele of a critical component of the phototransduction cascade, phospholipase C-&beta;, encoded by the norpA gene, to render the flies blind and also use the Gal4-UAS transcriptional activator system to drive expression of ChR2 in the Foma-1 neurons. Individual flies are placed on a small platform surrounded by blue LEDs. When the LEDs are illuminated, the flies quickly take-off into flight, in a manner similar to visually driven loom-escape behavior. We believe that this technique can be easily adapted to examine other behaviors in freely moving flies.

Optogenetic Stimulation of Escape Behavior in Drosophila melanogaster

Genetically encoded optogenetic tools enable noninvasive manipulation of specific neurons in the Drosophila brain. Such tools can identify neurons whose activation is sufficient to elicit or suppress particular behaviors. Here we present a method for activating Channelrhodopsin2 that is expressed in targeted neurons in freely walking flies.

で逃避行動のOptogenetic刺激<em&gt;キイロショウジョウバエ</em

A  growing number of genetically encoded tools are becoming available that allow non-invasive  manipulation of the neural activity of specific neurons in ...

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;

における自発運動量の測定<em&gt;キイロショウジョウバエ</em

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

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 ...

High-resolution Quantification of Odor-guided Behavior in Drosophila melanogaster Using the Flywalk Paradigm

In their natural environment, insects such as the vinegar fly Drosophila melanogaster are bombarded with a huge amount of chemically distinct odorants. To complicate matters even further, the odors detected by the insect nervous system usually are not single compounds but mixtures whose composition and concentration ratios vary. This leads to an almost infinite amount of different olfactory stimuli which have to be evaluated by the nervous system.
To understand which aspects of an odor stimulus determine its evaluation by the fly, it is therefore desirable to efficiently examine odor-guided behavior towards many odorants and odor mixtures. To directly correlate behavior to neuronal activity, behavior should be quantified in a comparable time frame and under identical stimulus conditions as in neurophysiological experiments. However, many currently used olfactory bioassays in Drosophila neuroethology are rather specialized either towards efficiency or towards resolution.
Flywalk, an automated odor delivery and tracking system, bridges the gap between efficiency and resolution. It allows the determination of exactly when an odor packet stimulated a freely walking fly, and to determine the animal&#180;s dynamic behavioral reaction.

High-resolution Quantification of Odor-guided Behavior in Drosophila melanogaster Using the Flywalk Paradigm

The automated tracking system Flywalk is used for high-resolution quantification of odor-guided behavior in Drosophila melanogaster.

臭気誘導挙動の高分解能定量<em&gt;キイロショウジョウバエ</em&gt;使用<em&gt; Flywalk</em&gt;パラダイム

In their natural environment, insects such as the vinegar fly Drosophila melanogaster are bombarded with a huge amount of chemically distinct odorants. To ...

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 ...

High Throughput Assay to Examine Egg-Laying Preferences of Individual Drosophila melanogaster

Recently, egg-laying preference of Drosophila has emerged as a genetically tractable model to study the neural basis of simple decision-making processes. When selecting sites to deposit their eggs, female flies are capable of ranking the relative attractiveness of their options and choosing the &#34;greater of two goods.&#34; However, most egg-laying preference assays are not practical if one wants to take a systematic genetic screening approach to search for the circuit basis underlying this simple decision-making process, as they are population-based and laborious to set up. To increase the throughput of studying of egg-laying preferences of single females, we developed custom chambers that each can simultaneously assay egg-laying preferences of up to thirty individual flies as well as a protocol that ensures each female has a high egg-laying rate (so that their preference is readily discernable and more convincing). Our approach is simple to execute and produces very consistent results. Additionally, these chambers can be equipped with different attachments to allow video recording the egg-laying animals and to deliver light for optogenetics studies. This article provides the blueprints for fabricating these chambers and the procedure for preparing the flies to be assayed in these chambers.

High Throughput Assay to Examine Egg-Laying Preferences of Individual Drosophila melanogaster

This protocol describes a high throughput assay for testing egg-laying preferences of Drosophila melanogaster at single-animal resolution. This assay provides a simple, efficient, and scalable platform to identify genes and circuit components that control a simple decision-making process.

個々の産卵環境を調べるために、ハイスループットアッセイ<em&gt;キイロショウジョウバエ</em

Recently, egg-laying preference of Drosophila has emerged as a genetically tractable model to study the neural basis of simple decision-making processes. ...

Foraging Path-length Protocol for Drosophila melanogaster Larvae

The Drosophila melanogaster larval path-length phenotype is an established measure used to study the genetic and environmental contributions to behavioral variation. The larval path-length assay was developed to measure individual differences in foraging behavior that were later linked to the foraging gene. Larval path-length is an easily scored trait that facilitates the collection of large sample sizes, at minimal cost, for genetic screens. Here we provide a detailed description of the current protocol for the larval path-length assay first used by Sokolowski. The protocol details how to reproducibly handle test animals, perform the behavioral assay and analyze the data. An example of how the assay can be used to measure behavioral plasticity in response to environmental change, by manipulating feeding environment prior to performing the assay, is also provided. Finally, appropriate test design as well as environmental factors that can modify larval path-length such as food quality, developmental age and day effects are discussed.

Foraging Path-length Protocol for Drosophila melanogaster Larvae

We provide a detailed protocol for a Drosophila melanogaster foraging path-length assay. We discuss the preparation and handling of test animals, how to perform the assay and analyze the data.

パスの長さのプロトコルを採餌<em&gt;キイロショウジョウバエ</em&gt;幼虫

The Drosophila melanogaster larval path-length phenotype is an established measure used to study the genetic and environmental contributions to behavioral ...

The CApillary FEeder Assay Measures Food Intake in Drosophila melanogaster

For most animals, feeding is an essential behavior for securing survival, and it influences development, locomotion, health and reproduction. Ingestion of the right type and quantity of food therefore has a major influence on quality of life. Research on feeding behavior focuses on the underlying processes that ensure actual feeding and unravels the role of factors regulating internal energy homeostasis and the neuronal bases of decision-making. The model organism Drosophila melanogaster, with its great variety of genetically traceable tools for labeling and manipulating single neurons, allows mapping of neuronal networks and identification of molecular signaling cascades involved in the regulation of food intake. This report demonstrates the CApillary FEeder assay (CAFE) and shows how to measure food intake in a group of flies for time spans ranging from hours to days. This easy-to-use assay consists of glass capillaries filled with liquid food that flies can freely access and feed on. Food consumption in the assay is accurately determined using simple measurement tools. Herein we describe step-by-step the method from setup to successful execution of the CAFE assay, and provide practical examples to analyze the food intake of a group of flies under controlled conditions. The reader is guided through possible limitations of the assay, and advantages and disadvantages of the method compared to other feeding assays in D. melanogaster are evaluated.

The CApillary FEeder Assay Measures Food Intake in Drosophila melanogaster

The CApillary FEeder (CAFE) assay is a simple, budget-friendly, highly reliable method for investigating mechanisms underlying food intake. Used with the highly versatile genetic model organism Drosophila melanogaster, it provides a powerful means of gaining new insights into regulatory mechanisms of food intake.

キャピラリーフィーダーアッセイにおいて食物摂取を測定します<em&gt;キイロショウジョウバエ</em

For most animals, feeding is an essential behavior for securing survival, and it influences development, locomotion, health and reproduction. Ingestion of ...

Assay Development for High Content Quantification of Sod1 Mutant Protein Aggregate Formation in Living Cells

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease that can be caused by inherited mutations in the gene encoding copper-zinc superoxide dismutase 1 (SOD1). The structural instability of SOD1 and the detection of SOD1-positive inclusions in familial-ALS patients supports a potential causal role for misfolded and/or aggregated SOD1 in ALS pathology. In this study, we describe the development of a cell-based assay designed to quantify the dynamics of SOD1 aggregation in living cells by high content screening approaches. Using lentiviral vectors, we generated stable cell lines expressing wild-type and mutant A4V SOD1 tagged with yellow fluorescent protein and found that both proteins were expressed in the cytosol without any sign of aggregation. Interestingly, only SOD1 A4V stably expressed in HEK-293, but not in U2OS or SH-SY5Y cell lines, formed aggregates upon proteasome inhibitor treatment. We show that it is possible to quantify aggregation based on dose-response analysis of various proteasome inhibitors, and to track aggregate-formation kinetics by time-lapse microscopy. Our approach introduces the possibility of quantifying the effect of ALS mutations on the role of SOD1 in aggregate formation as well as screening for small molecules that prevent SOD1 A4V aggregation.

We describe a method to quantify the aggregation of misfolded proteins. Our protocol details lentiviral induced stable cell line generation, automated confocal imaging, and image analysis of protein aggregates. As an illustrative application, we studied the effect of small molecules in promoting SOD1 aggregation in a time- and dose-dependent manner.

Sod1 突然変異体蛋白質集計における細胞の高いコンテンツ定量化のための分析法の開発

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease that can be caused by inherited mutations in the gene encoding copper-zinc ...

社会回避での定量化のための簡単​​なアッセイ

社会回避での定量化のための簡単なアッセイ