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.

NOTA: Questo protocollo descrive la preparazione (sezioni 1 - 3), esecuzione (sezione 4), e le analisi (sezione 4) di saggi sazietà 2-D. Quindi, utilizzando la stimolazione dopaminergica come esempio, la sezione 5 mostra come combinare la stimolazione thermogenetic con saggi sazietà 2-D per indurre ipersessualità. Sezione 6 descrive 3 modi per verificare i risultati dei dosaggi sazietà 2-D. Infine, la sezione 7 mostra come misurare il recupero delle unità di accoppiamento nei moscerini maschi. <p class="jove_ti...

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Stati motivazionali possono essere saziati, mantenuti e recuperati 34. Vi presentiamo un test sazietà 2-D che misura in modo rapido e robusto tutti questi aspetti di accoppiamento auto in tempo reale. Questo test apre la possibilità di utilizzare mosca advanced manipolazioni genetiche per studiare le componenti molecolari e circuitali di un comportamento motivato. Il test si basa sulla capacità di sazietà del maschio in tribunale con successo e copulare, e di inte...

Il lavoro in Drosophila ha fornito una visione profonda e pionieristico in molti fenomeni biologici, compresa la natura del gene 1, i principi dello sviluppo embrionale 2, ritmi circadiani 3, e lo sviluppo e il cablaggio del sistema nervoso 4, 5, 6. La motivazione resta molto meno ben compreso che questi fenomeni, forse a causa delle limitazioni sui sistemi che sono stati studiati finora. La motivazione e la mosca è studiato principalmente nel contesto della fame, che presenta molte sfide a causa della loro irrisorio assunzione di cibo per attacco di alimentazione e di esoscheletro che esclude i segni evidenti di deposizione di grasso. Di conseguenza, vi è la necessità di ampliare i sistemi utilizzati per studiare motivazione in volo. Descriviamo un quadro comportamentale per lo studio di azionamento accoppiamento inDrosophila. Questo sistema sfrutta gli strumenti neurogenetic in volo e l&#39;accessibilità 7, 8, 9, 10, 11, 12 e la connettoma putativo del suo circuito dimorfismo sessuale 8, 13. Inoltre, gran parte della innata 14, 15, 16, 17, 18, 19, 20, 21 e ho imparato 22, 23, 24 circuiti senso-motorio che controlla il corteggiamento è stato elaborato in dettaglio, offrendo una rara opportunitàper individuare il nodo circuitale esatta su cui incide motivazione. Abbiamo recentemente riportato che, nel volo, come negli esseri umani, i livelli di dopamina sono fondamentali per unità di accoppiamento 25, 26, 27. Abbiamo ottenuto l&#39;accesso genetica al relativo dettagliata a livello di circuito molecular- e che producono la dopamina e la ricezione di neuroni al volo, facilitando l&#39;analisi di questo fenomeno conservato utilizzando i saggi che descriviamo qui 25. Aggiungiamo ai test comportamentali nei Zhang et al. 25 una nuova arena comportamentale piatta che permette di video di punteggio, che noi chiamiamo un test di sazietà a 2 dimensioni (2-D), un importante miglioramento rispetto ai metodi precedenti. Di conseguenza, il nuovo saggio è più scalabile e quantificabile e quindi più adatto per schermi genetiche di geni e neuroni coinvolti nella motivazione. Usiamo questo nuovo test, insieme a saggi di corteggiamento e neurogemanipolazioni tiche, per dimostrare come misurare e modificare unità di accoppiamento in volo.

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			<td height="21" style="height:21px;width:216px;">1/16 inch clear acrylic</td>
			<td style="width:119px;">McMaster-Carr</td>
			<td style="width:119px;">8589K12</td>
			<td style="width:743px;">Used to make arenas; see Supplemental Material 1 for designs.</td>
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		<tr height="21">
			<td height="21" style="height:21px;">1/8 inch clear acrylic</td>
			<td style="width:119px;">McMaster-Carr</td>
			<td>8589K42</td>
			<td>Used to make arenas; see Supplemental Material 1 for designs.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">3/16 inch clear acrylic</td>
			<td style="width:119px;">McMaster-Carr</td>
			<td>8560K219</td>
			<td>Used to make arenas; see Supplemental Material 1 for designs.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">1/32 inch black delrin</td>
			<td style="width:119px;">McMaster-Carr</td>
			<td>8575K132</td>
			<td>Used to make arenas; see Supplemental Material 1 for designs.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hex screws, 1 inch long (50x)</td>
			<td style="width:119px;">McMaster-Carr</td>
			<td>92314A115&#160;</td>
			<td>Used to make arenas. Can be replaced by 3/4 inch screws (92314A113, McMaster-Carr) for 32-chamber arenas.</td>
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		<tr height="21">
			<td height="21" style="height:21px;">Thumb nuts (25x)</td>
			<td style="width:119px;">McMaster-Carr</td>
			<td>92741A100</td>
			<td>Used to make arenas. Can be replaced by regular hex nuts (90480A005, McMaster-Carr).</td>
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		<tr height="21">
			<td height="21" style="height:21px;">Camcorder</td>
			<td style="width:119px;">Canon</td>
			<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

Nonostante decenni di ricerca, le basi neuronali e molecolari degli stati motivazionali rimangono misteriose. Abbiamo recentemente sviluppato un nuovo, riduzionista, e il sistema scalabile per un&#39;indagine approfondita della motivazione utilizzando l&#39;unità di accoppiamento del maschio Drosophila melanogaster (Drosophila), i metodi per la quale abbiamo dettaglio qui. Il paradigma comportamentale centra sulla constatazione che l&#39;unità di accoppiamento maschio diminuisce la fertilità a fianco nel corso di accoppiamenti ripetuti e recupera oltre ~ 3 d. In questo sistema, gli strumenti potenti neurogenetici disponibili nei fly convergono con l&#39;accessibilità genetica e schema elettrico putativo disponibile per comportamento sessuale. Questa convergenza consente un rapido isolamento e l&#39;interrogatorio di piccole popolazioni neuronali con specifiche funzioni motivazionali. Qui si dettaglio la progettazione e l&#39;esecuzione del test sazietà che viene utilizzato per misurare e modificare la motivazione di corteggiamento nel moscerino maschio. l&#39;utilizzo di questotest, abbiamo anche dimostrato che rasoterra accoppiamento maschio può essere superata attraverso la stimolazione dei neuroni dopaminergici. Il saggio sazietà è semplice, economico e robusto per le influenze di background genetico. Ci aspettiamo che il saggio di sazietà per generare molte nuove intuizioni nella neurobiologia degli stati motivazionali.

Per caratterizzare unità di accoppiamento Drosophila, 3 giorni di età, maschi WT Canton-S sono stati testati in un saggio sazietà 2-D. Nel corso del test (4.5 h), i maschi si accoppiano una media di 4,8 ± 0,3 (media ± errore standard della media, SEM) volte. Accoppiamenti iniziano soprattutto nei primi 2 h (78%) (Figura 6A, 6B) e diventano meno frequenti come il saggio progredisce (Figura 6A, 6B). Questa diminuzione non è dovuta alla manca...

Watch this Scientific Journal Video about Measuring and Altering Mating Drive in Male Drosophila melanogaster at JoVE.com

Measuring and Altering Mating Drive in Male Drosophila melanogaster

Questo articolo descrive un test comportamentale che utilizza unità di accoppiamento maschio in Drosophila melanogaste R per studiare la motivazione. Utilizzando questo metodo, i ricercatori possono utilizzare mosca avanzate tecniche neurogenetiche per scoprire i meccanismi genetici, molecolari e cellulari che sono alla base questa motivazione.

Misurazione e l&#39;alterazione di accoppiamento Drive a Maschio<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, ...

Research

JoVE Journal

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

Registrazioni Sensillum singolo in Insetti Drosophila melanogaster E Anopheles gambiae</em

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

Ricerca

Neuroscienze

Optogenetic Stimulation of Escape Behavior in Drosophila melanogaster

Optogenetic Stimulation of Escape Behavior in Drosophila melanogaster

Stimolazione Optogenetic del comportamento in Fuga<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

Determinazione del locomotore attività spontanea in<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

Assay Semplice per la quantificazione della Social Prevenzione in<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

Tintoria Insetti per comportamentali saggi: il comportamento di accoppiamento di anestetizzati<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

Condizioni che influiscono spazio sociale in<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

Foraggiamento protocollo percorso di lunghezza per<em&gt; Drosophila melanogaster</em&gt; Le larve

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

Il capillare alimentatore Assay Misure assunzione di cibo in<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

Analisi metabolica di Drosophila melanogaster Larval e cervello adulto

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

Valutazione morfologica e funzionale degli assoni e delle loro sinapsi durante la morte di Axon in Drosophila melanogaster

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