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.

MERK: Denne protokollen beskriver forberedelsene (§§ 1 - 3), gjennomføring (kapittel 4), og analyse (§ 4) 2-D satiety analyser. Deretter bruker dopaminerg stimulering som eksempel § 5 viser hvordan du kan kombinere thermogenetic stimulering med 2-D satiety analyser for å indusere hyperseksualitet. § 6 beskriver 3 måter å bekrefte resultatene av 2-D satiety analyser. Til slutt, § 7 viser hvordan man kan måle utvinning av parring stasjonen i mannlige fluer. 1. Fabricating 8- og 32-kam...

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Motiverende stater kan bli mett, vedlikeholdt, og gjenvunnet 34. Vi presenterer en 2-D satiety analysen som raskt og robust måler alle disse aspektene ved parring stasjonen i fly. Denne analysen åpner opp muligheten for å bruke avanserte flue genetiske manipulasjoner for å studere de molekylære og krets komponenter av en motivert atferd. Satiety analysen er avhengig av mannens evne til å retten og parre seg, og å avslutte copulations på riktig tidspunkt. Selv ...

Arbeid i Drosophila har gitt dype og banebrytende innsikt i mange biologiske fenomener, herunder arten av genet 1, prinsipper for embryoutvikling 2, døgnrytme 3, og utvikling og kabling av nervesystemet 4, 5, 6. Motivasjon er fortsatt langt mindre forstått enn disse fenomenene, kanskje på grunn av begrensninger på de systemene som har blitt studert så langt. Motivasjon i fly er primært studert i sammenheng med sult, som presenterer mange utfordringer på grunn av deres forsvinnende liten matinntak per fôring bout og exoskeleton som utelukker klare tegn på fett deponering. Følgelig er det et behov for å utvide de systemer som benyttes for å studere motivasjon i fly. Vi beskriver en atferds rammeverk for studiet av parring stasjonen iDrosophila. Dette systemet utnytter de neurogenetic verktøyene i fly så vel som tilgjengeligheten 7, 8, 9, 10, 11, 12 og den antatte connectome av dens seksuelt dimorf kretsen 8, 13. I tillegg er mye av det medfødte 14, 15, 16, 17, 18, 19, 20, 21 og lært 22, 23, 24 sensoriske-motor kretser kontrollerende frieri er utarbeidet i detalj, og gir en sjelden mulighetå finne den eksakte kretsen node hvorpå motivasjon treffer. Vi har nylig rapportert at, i fly, som hos mennesker, dopamin-nivåene er sentrale for mating stasjon 25, 26, 27. Vi har fått genetisk tilgang til relevant dopamin-produserende og motta nevroner i fly, tilrettelegging detaljert molekylær- og krets-nivå analyser av dette bevart fenomenet ved hjelp av analysene vi beskriver her 25. Vi legger til atferdsanalyser i Zhang et al. 25 en ny flat atferds arena som gjør at video å score, som vi kaller en to-dimensjonale (2D) satiety analyse, en viktig forbedring i forhold til tidligere metoder. Følgelig er den nye analysen mer skalerbar og kvantifiserbar, og derfor mer egnet for genetiske skjermer av gener og neuroner som er involvert i motivasjon. Vi bruker denne nye analysen, sammen med frieri analyser og neurogemagnetiske manipulasjoner, for å demonstrere hvordan man kan måle og endre parring stasjonen i fly.

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			<td height="21" style="height:21px;width:216px;">1/16 inch clear acrylic</td>
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			<td height="21" style="height:21px;">3/16 inch clear acrylic</td>
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			<td>Used to make arenas; see Supplemental Material 1 for designs.</td>
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			<td height="21" style="height:21px;">1/32 inch black delrin</td>
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			<td>Used to make arenas; see Supplemental Material 1 for designs.</td>
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			<td style="width:119px;">McMaster-Carr</td>
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			<td height="21" style="height:21px;">Camcorder</td>
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			<td>Vixia HF R700</td>
			<td>Can be replaced by any consumer comcorder.</td>
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measuring and altering mating drive in male drosophila melanogaster

Til tross for flere tiår med etterforskning, de nevronale og molekylære grunnlaget for motiverende stater forblir mystisk. Vi har nylig utviklet en ny, reduksjonistisk, og skalerbart system for grundig undersøkelse av motivasjonen med parring stasjonen på mannlig bananflue (Drosophila), metodene som vi detalj her. Den atferds paradigmet sentre på funn at mannlige parring kjøretur synker sammen fruktbarhet i løpet av gjentatte copulations og gjen løpet ~ 3 d. I dette systemet, de kraftige neurogenetic verktøy tilgjengelig i flue konvergere med den genetiske tilgjengelighet og antatte koblingsskjema tilgjengelig for seksuell atferd. Dette konvergens tillater rask isolasjon og avhør av små nevronale populasjoner med spesifikke motiverende funksjoner. Her har vi detalj utforming og gjennomføring av metthet analyse som brukes til å måle og endre frieri motivasjon i den mannlige fly. Ved hjelp av denneanalysen, viser vi også at lav mannlige parring stasjonen kan overvinnes ved å stimulere dopaminerge nevroner. Satiety analysen er enkel, rimelig og robust til påvirkninger av genetisk bakgrunn. Vi forventer at satiety analyse for å generere mange nye innsikt i nevrobiologi motiverende stater.

For å karakterisere Drosophila parring stasjon, tre dager gamle, WT Canton-S-hanner ble testet i en to-D metthetsfølelse analysen. I løpet av analysen (4,5 timer), hanner parer et gjennomsnitt på 4,8 ± 0,3 (middelverdi ± standardavvik av middel, SEM) ganger. Parr initiere for det meste i de første 2 timer (78%) (Figur 6A, 6B) og blir mindre hyppig som analysen utvikler seg (figur 6A, 6B). Denne reduksjonen er ikke på grunn av mangel på ...

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Measuring and Altering Mating Drive in Male Drosophila melanogaster

Denne artikkelen beskriver en atferdsanalyse som bruker mannlige parring stasjonen i Drosophila melanogaste r å studere motivasjon. Ved hjelp av denne metoden, kan forskerne utnytte avansert flue neurogenetic teknikker for å avdekke genetiske, molekylære og cellulære mekanismer som ligger til grunn for denne motivasjonen.

Måle og endre Mating Drive i 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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Neuroscience

Measuring and Altering Mating Drive in Male Drosophila melanogaster

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

The sense of smell is essential for insects to find foods, mates, predators, and oviposition sites3. Insect olfactory sensory neurons (OSNs) are enclosed ...

Optogenetic Stimulation of Escape Behavior in Drosophila melanogaster

Optogenetic Stimulation of Escape Behavior in Drosophila melanogaster

Optogenetic Stimulering av Escape Behavior i<em&gt; Drosophila melanogaster</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

Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster

Fastsettelse av Spontan Locomotor aktivitet i<em&gt; Drosophila melanogaster</em

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

Straightforward Assay for Quantification of Social Avoidance in Drosophila melanogaster

Straightforward Assay for Quantification of Social Avoidance in Drosophila melanogaster

Enkel analyse for Kvantifisering av Social Avhold 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 ...

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

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

Farging Insekter for Behavioral Analyser: den Mating Behavior av bedøvd<em&gt; Drosophila</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

Conditions Affecting Social Space in Drosophila melanogaster

Forhold som påvirker sosiale rom i<em&gt; Drosophila melanogaster</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 ...

Foraging Path-length Protocol for Drosophila melanogaster Larvae

Foraging Path-length Protocol for Drosophila melanogaster Larvae

Foraging Sti lengde Protokoll for<em&gt; Drosophila melanogaster</em&gt; Larver

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

The CApillary FEeder Assay Measures Food Intake in Drosophila melanogaster

Kapillær mater analysen Måler matinntak i<em&gt; Drosophila melanogaster</em

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

Metabolic Analysis of Drosophila melanogaster Larval and Adult Brains

Metabolic Analysis of Drosophila melanogaster Larval and Adult Brains

Metabolsk analyse av Drosophila melanogaster Larval og voksen hjerner

This protocol describes a method for measuring the metabolism in Drosophila melanogaster larval and adult brains. Quantifying metabolism in whole organs ...

Morphological and Functional Evaluation of Axons and their Synapses during Axon Death in Drosophila melanogaster

Morphological and Functional Evaluation of Axons and their Synapses during Axon Death in Drosophila melanogaster

Morfologisk og funksjonell evaluering av axoner og deres synapser under Axon Death i Drosophila melanogaster

Axon degeneration is a shared feature in neurodegenerative disease and when nervous systems are challenged by mechanical or chemical forces. However, our ...