Name: measuring and altering mating drive in male drosophila melanogaster
Uploaded: 2017-02-15T14:00:00
Description: Watch this Scientific Journal Video about Measuring and Altering Mating Drive in Male Drosophila melanogaster at JoVE.com

The authors thank Mike Crickmore, Dragana Rogulja, and Michelle Frank for comments on the manuscript. Pavel Gorelik provided technical support for manufacturing the behavioral arenas. This work was conducted in Mike Crickmore's lab and is also supported by the Whitehall Foundation (Principal Investigator: Dragana Rogulja). S.X.Z. is a Stuart H.Q. and Victoria Quan Fellow at Harvard Medical School.

LET OP: Dit protocol beschrijft de bereiding (secties 1-3), uitvoering (hoofdstuk 4), en de analyse (hoofdstuk 4) van 2-D verzadiging assays. Dan, met dopaminerge stimulatie als voorbeeld, afdeling 5 laat zien hoe thermogenetische stimulatie combineren met 2-D verzadiging testen voor hypersexuality induceren. Hoofdstuk 6 beschrijft 3 manieren om de resultaten van 2-D verzadiging assays verifiëren. Tot slot, Hoofdstuk 7 laat zien hoe het herstel van de paring drive in mannelijke vliegt meten. ...

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Motivational toestanden kunnen worden verzadigd, onderhouden en hersteld 34. We presenteren een 2-D verzadiging test die snel en krachtig meet al deze aspecten van paring aandrijving in de vlieg. Deze test opent de mogelijkheid om geavanceerde fly genetische manipulatie de moleculaire en circuitcomponenten van een gemotiveerd gedrag te bestuderen. De verzadiging test is gebaseerd op het vermogen van de man om met succes de rechtbank en copuleren, en copulaties op het ...

Werk in Drosophila heeft diepe en baanbrekende inzicht gegeven in vele biologische fenomenen, zoals de aard van het gen 1, beginselen van embryonale ontwikkeling 2, circadiane ritmes 3, en de ontwikkeling en bedrading van het zenuwstelsel 4, 5, 6. Motivatie blijft veel minder goed begrepen dan deze verschijnselen, misschien vanwege de beperkingen van de systemen die tot nu toe zijn onderzocht. Motivatie in de vlieg wordt vooral onderzocht in het kader van de honger, die vele uitdagingen presenteert als gevolg van hun verwaarloosbaar klein voedselinname per voeding wedstrijd en exoskeleton die duidelijke tekenen van vetopslag in de weg staat. Bijgevolg is er een behoefte om de systemen voor het bestuderen motivatie in de vlieg breiden. We beschrijven een gedrags raamwerk voor de studie van de paring drive inDrosophila. Dit systeem maakt gebruik van de neurogenetische instrumenten in de vlieg en de toegankelijkheid 7, 8, 9, 10, 11, 12 en de vermeende connectoom van de sexueel dimorfe schakelingen 8, 13. Bovendien, veel van de aangeboren 14, 15, 16, 17, 18, 19, 20, 21 en geleerde 22, 23, 24 sensomotorische Schakelingen courtship is uitgewerkt, die een zeldzame kansom de exacte circuit knooppunt waarop motivatie botst lokaliseren. We hebben onlangs gemeld dat, in de vlieg, zoals bij mensen, dopamine niveaus centraal paring aandrijving 25, 26, 27. We hebben genetische toegang tot de relevante dopamine-producerende en het ontvangen van neuronen in de vlieg, het vergemakkelijken van gedetailleerde moleculair-en circuit-niveau opgedaan analyses van deze geconserveerde fenomeen met behulp van de testen beschrijven we hier 25. Voegt men de gedrags assays Zhang et al. 25 een nieuwe flatscreen gedrags-arena die video scoren, dat we een 2-dimensionale (2-D) verzadiging assay, een belangrijke verbetering ten opzichte van eerdere methoden bellen mogelijk maakt. Bijgevolg is de nieuwe assay is schaalbaar en meetbaar, en dus meer geschikt voor genetische screens van genen en neuronen betrokken bij motivatie. We maken gebruik van deze nieuwe test, samen met de verkering assays en neurogesche manipulaties, om aan te tonen hoe om te meten en te veranderen paring rijden in de vlieg.

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measuring and altering mating drive in male drosophila melanogaster

Ondanks tientallen jaren van onderzoek, het neuronale en moleculaire basis van motivationele toestanden blijven mysterieus. We hebben onlangs een nieuwe, reductionistische en schaalbaar systeem voor een diepgaand onderzoek van de motivatie met behulp van de paring aandrijving van mannelijke Drosophila melanogaster (Drosophila), de methoden waarvoor Hier hebben we detail. De behavioral paradigma centra op de bevinding dat mannetje mating schijf afneemt langs de vruchtbaarheid in de loop van herhaalde copulaties en herstelt meer dan ~ 3 d. In dit systeem is de krachtige neurogenetische gereedschap in de vlieg samen met de genetische toegankelijkheid en vermeende aansluitschema vindt seksueel gedrag. Deze convergentie maakt een snelle isolatie en ondervraging van kleine neuronale populaties met specifieke motivationele functies. Hier hebben we detail het ontwerp en de uitvoering van de verzadiging test die wordt gebruikt voor het meten en veranderen verkering motivatie in de mannelijke vliegen. gebruik van dezeassay, we tonen ook aan dat een lage mannetje mating drive kan worden overwonnen door het stimuleren van dopaminerge neuronen. De verzadiging test is eenvoudig, betaalbaar en robuust invloeden van de genetische achtergrond. Wij verwachten dat de verzadiging test om veel nieuwe inzichten in de neurobiologie van motivationele toestanden te genereren.

Karakterisering Drosophila paring aandrijving, 3 dagen oude, WT Canton-S mannetjes werden getest in een 2-D verzadiging assay. In de loop van de test (4,5 h), mannetjes mate gemiddeld 4,8 ± 0,3 (gemiddelde ± standaardfout van het gemiddelde, SEM) keer. Paringen initiëren meestal in de eerste 2 uur (78%) (figuur 6A, 6B) en minder frequent als test vordert (figuur 6A, 6B). Deze daling is niet te wijten aan het gebrek aan passende partners (74% ...

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Measuring and Altering Mating Drive in Male Drosophila melanogaster

Dit artikel beschrijft een gedrags-test die mannetje mating drive in Drosophila melanogaste r om de motivatie te bestuderen gebruikt. Met deze methode kunnen onderzoekers gebruiken geavanceerde fly neurogenetische technieken om de genetische, moleculaire en cellulaire mechanismen die ten grondslag liggen aan deze motivatie ontdekken.

Het meten en de wijziging van Mating Drive in Male<em&gt; Drosophila melanogaster</em

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

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Measuring and Altering Mating Drive in Male Drosophila melanogaster

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