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

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

1. Utrustning och reagenser Skapad Egen (se Förteckning över material för andra) <ol><li> Förbered en Drosophila kallt anestesi apparat för att utföra flyga arbete. <ol><li> Skär ett poröst polyetenark för att täcka en liten plastlåda (12,7 cm lång, 10,2 cm bred), typiskt den övre luckan på en pipettspetsar låda (12,7 cm lång, 10,2 cm bred, 3,8 cm djup). </li><li> Fyll lådan med krossad is, täck med den porösa polyetenark. </li></ol></li><li> Förbered en T-labyrint apparat; anpassa anordningen som ...

<ol>
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Detta protokoll beskriver ett detaljerat förfarande för undvikande analysen sociala. Canton-S kommer bara undvika en flaska där flyger har tidigare varit mekaniskt stressad, och att kön och antal responders påverkar inte den sociala undvikande prestanda. Men genetisk bakgrund av responders har ett stort inflytande. Följande är flera viktiga steg för att utföra detta experiment framgångsrikt: 1) alltid överföra flugorna 2 tim före försöket och se till att inte störa sin omgivn...

Målet med denna metod är att enkelt kvantifiera en ny aspekt av enkla socialt beteende i Drosophila melanogaster, oberoende av uppvaktning och aggression. Social interaktion är avgörande för en god utveckling och hälsa individer inom ett samhälle, samt funktionaliteten hos en social grupp som helhet. Den höga komplexiteten i dessa interaktioner kräver stora provstorlekar och ett system som gör det möjligt för förenkling av beteende som de genetiska och neurala grunderna för socialt beteende fortfarande dåligt kända. Drosophila melanogaster är en kraftfull genetisk modell som kan användas för att identifiera genetiska och neurala baser av sociala interaktioner. Indeed, D. melanogaster har en repertoar av komplexa sociala beteenden och några direkta mätningar av socialisering har redan gjorts 1-7. Men de flesta av dessa ansträngningar har fokuserat på relativt komplexa sociala beteenden, såsom aggressiva interaktion 3,6, olika aspekter av uppvaktning 3,8-12, och hur sociala erfarenheter påverkar andra beteenden såsom inlärning, eller dygnsrytm 13-17. Dessutom är många av dessa analyser förlitar sig på att analysera komplexa interaktionsmönster grupper av flugor, med hjälp av video tracking och datorprogram för att analysera den resulte överflöd av data. Sådana analyser är ovärderliga, och leda till nya viktiga insikter såsom dynamik fly-fly interaktioner i grupperna 7. En begränsning är dock otillgänglighet av dessa analyser till samhället i stort, och den begränsade kunskapen om mekanismerna bakom erkännande av andra. Med andra ord, är grunden för utsläpp av en signal från en individ och dess erkännande av en annan fortfarande dåligt kända 18. Däremot flyger också uppvisar en enkel beteende, socialt undvikande, där individer röra sig bort från en signal som skickas av stressade flugor: D. Melanogaster Stress odorant eller DSO 19. I en hög genomströmning analys, kan detta beteende kvantifieras som undvikande av en stress signal som skickas av andra flugor, eller socialt undvikande 19. Flugor placeras i en T-labyrint apparat och ges möjlighet att undvika en flaska innehållande DSO. Med hjälp av denna analys, blev CO2 visade sig vara en komponent i DSO, och en del av neurala kretsar som krävs för att svara på CO2 dissekerades 19. Analysen sociala undvikande presenteras här liknar konceptuellt till de enkla beteendeanalyser som utvecklats i Seymour Benzer laboratorium som möjlig generationer av forskare för att dissekera komplexa beteenden 20. Analys av social undvikande kan genomföras kostnadseffektivt med hjälp av T-labyrinten analys, vilket möjliggör mer utbredd studie av socialt beteende. Till exempel med hjälp av denna analys som vi nyligen visat att olika genetiska risker för autism har kontrasterande effekter i socialt behavior analyser. Mutanter för en kandidat gen för autism - neurobeachin 21,22 - nuvarande brister både i det sociala rummet (beskrivs på annat håll 23) och socialt undvikande 24. Onormal dopaminerga signalering föreslås också att spela en roll i etiologin för autism hos människor 25,26. I motsats till de resultat som erhållits med neurobeachin, fann vi att social undvikande prestanda var opåverkad av ökad eller minskad nivåer av Drosophila vesikulär Monoamin Transporter (VMAT) i dopaminerga celler, även om det sociala rummet var direkt korrelerad till dessa nivåer av VMAT 27. De kontrasterande resultat som erhållits med neurobeachin och VMAT stryker möjligheten att identifiera olika former av asocialt beteende och därmed olika underliggande neurala circuitries modulerar responsen på andra.

<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.

Undvikande analysen sociala är en robust testet kvantifiera förmågan hos Drosophila melanogaster att känna igen en stresssignal (DSO) som avges av andra flugor, och därmed bedöma en aspekt av sociala interaktioner. Analysen utförs med hjälp av en apparat som vanligen används i olika beteendeanalyser som kallas en T-labyrint, som presenterar flugorna med ett val mellan två olika alternativ - vänster eller höger 19,28-31. I detta fall, är effektiviteten vid vilken flugorna undvika DSO kvan...

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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.

Enkel analys för kvantifiering av social Undvikande i<em&gt; Drosophila melanogaster</em

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

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Straightforward Assay for Quantification of Social Avoidance in Drosophila melanogaster

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