Name: measuring and altering mating drive in male drosophila melanogaster
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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.

NOT: Bu protokol hazırlanması (Bölüm 1-3) açıklar, yürütme (Bölüm 4), ve analiz 2-D tokluk tahlillerin (Bölüm 4). Sonra, bir örnek olarak dopaminerjik stimülasyon kullanarak, Bölüm 5 hiperseksüalite ikna etmek için 2-D tokluk deneyleri ile thermogenetic stimülasyon birleştirmek nasıl gösterir. Bölüm 6 2-D tokluk deneyleri sonuçları doğrulamak için 3 yolları anlatılmaktadır. Son olarak, Bölüm 7 erkek sinekler çiftleşme sürücünün kurtarma ölçmek için nasıl gösterir. <p class...

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Motivasyonel devletler, tok muhafaza ve 34 elde edilebilir. Biz hızlı ve sağlam sinek sürücüyü çiftleşme bu tüm yönleriyle ölçen bir 2-B tokluk test sunuyoruz. Bu deney, bir motive davranış moleküler ve devre elemanlarını okumak için gelişmiş uçucu genetik manipülasyonlar kullanılması olasılığını yaratır. tokluk tahlil erkeğin yeteneği başarıyla mahkemeye ve çiftleşmek ve uygun zamanda çiftleştirerek sonlandırmak için kullanır...

Drosophila iş geninin 1. doğası dahil olmak üzere birçok biyolojik olayların derinliklerine ve öncü fikir sağlamıştır, embriyonik gelişim 2, sirkadiyen ritimler 3 ve sinir sistemi 4, 5, 6 gelişmesi ve kablolama ilkeleri. Motivasyon belki de şimdiye kadar çalışılmıştır sistemlerde sınırlamalar, çok daha az iyi bu olayların daha anlaşılamamıştır. sinek Motivasyon öncelikle yağ depolanmasının açık belirtilerini engeller besleme maçın ve exoskeleton başına onların yok denecek kadar az gıda alımı için birçok zorluklarla karşı karşıya açlık bağlamında ele alınmıştır. Sonuç olarak, sinek motivasyon incelemek için kullanılan sistemler genişletmek için bir ihtiyaç vardır. Biz çiftleşme sürücünün çalışmasında bir davranış çerçeveyi tanımlamakDrosophila. Bu sistem, sinek gibi erişilebilirlik 7, 8, 9, 10, 11, 12 ve cinsi olarak dimorfik devresi 8, 13, varsayılan connectome içinde nörogenetik araçlar yararlanır. Buna ek olarak, çok doğal, 14, 15, 16, 17, 18, 19, 20, 21 ve 22, 23, 24 duyu-motor devre kontrol kur nadir bir fırsat sağlamaktadır ayrıntılı bir şekilde olmuştur öğrenilenmotivasyon çarptığı bunun üzerine tam devre düğümü bulun. Son zamanlarda sinek, insanlarda olduğu gibi, dopamin düzeyleri eşleşen tahrik 25, 26, 27 için merkezi bildirmiştir. Biz burada 25 tarif deneyleri kullanılarak bu korunmuş fenomenin analizi, ilgili dopamin üreten ve sinek nöronlar alma kolaylaştıran detaylı Molecular ve devre düzeyinde genetik erişim kazanmıştır. Bu Zhang ve arkadaşları davranış testleri ekle. Biz 2 boyutlu (2-D) tokluk testi, önceki yöntemlere göre önemli bir iyileşme çağrı, video puanlama, izin verir 25 yeni bir düz davranış arenada. Sonuç olarak, yeni tahlil daha ölçeklenebilir ve ölçülebilir ve motivasyon yer alan genlerin ve nöronların genetik ekranlar için bu nedenle daha uygundur. Bu kur analizleri ve neuroge ile birlikte bu yeni tahlil kullanımımanyetik manipülasyonlar, ölçmek ve sinek çiftleşme sürücüyü değiştirmek için nasıl göstermek için.

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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>
			<td style="width:119px;">McMaster-Carr</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;">Hex screws, 1 inch long (50x)</td>
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			<td>92314A115&#160;</td>
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			<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

soruşturmanın yıllardır rağmen, motivasyon devletlerin nöronal ve moleküler temelleri gizemli kalır. Biz son zamanlarda burada detay metotları olan erkek Drosophila melanogaster (Drosophila) çiftleşme sürücüyü kullanarak motivasyon derinlemesine soruşturulması için bir roman, indirgemeci ve ölçeklenebilir bir sistem geliştirdik. davranışsal paradigma erkek çiftleşme sürücü tekrarlanan çiftleştirerek boyunca doğurganlık yanında azalır ve ~ 3 d üzerinde kurtarır bulgu merkezleri. Bu sistemde, sinek mevcut güçlü neurogenetic araçlar genetik erişilebilirlik ve cinsel davranış için varsayılan bağlantı şemasına ile birleşir. Bu yakınlaşma hızlı izolasyon ve özel motivasyon fonksiyonları ile küçük nöronal popülasyonlarının sorgulama sağlar. Burada detay ölçmek ve erkek sinek kur motivasyonu değiştirmek için kullanılır tokluk testinin tasarımı ve yürütülmesi biz. Kullanımı BuTahlil, biz de düşük erkek çiftleşme sürücü dopaminerjik nöronların uyararak aşılabilir olduğunu göstermektedir. tokluk tahlil genetik arka plan etkilerine basit, hesaplı ve sağlamdır. Biz tokluk tahlil motivasyon devletler nörobiyolojisine içine birçok yeni anlayışlar üretmek için bekliyoruz.

Drosophila eşleşen tahrik karakterize etmek için, 3 günlük bir WT Canton-S erkek 2-B doyma deneyi ile test edildi. tahlil (4.5 saat) boyunca, erkekler 4.8 ± 0.3 ortalama arkadaşı kez (ortalama, SEM ± standart hata demek). Eşleştirmesi, ilk 2 saat boyunca (% 78) (Şekil 6A, 6B) &#39;de daha çok başlatmak ve deneyi (Şekil 6A, 6B) ilerledikçe sıklığı azalır. Bu azalma çiftleşme ortakları eksikliği (% 74 kadın tahlil boyunca...

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

Bu makale melanogaste motivasyon incelemek için r Drosophila erkek çiftleşme sürücüsünü kullanan bir davranış deneyi anlatır. Bu yöntemi kullanarak, araştırmacılar bu motivasyonu altında yatan genetik moleküler ve hücresel mekanizmaları ortaya çıkarmak için ileri sinek neurogenetic teknikler kullanabilirler.

Ölçme ve Male Değiştirme Çiftleşme Sürücü<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

Böcekler Tek Sensillum Kayıtlar Drosophila melanogaster Ve Anofel 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 ...

Optogenetic Stimulation of Escape Behavior in Drosophila melanogaster

Optogenetic Stimulation of Escape Behavior in Drosophila melanogaster

Yılında Escape Davranış Optogenetic Uyarım<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

Olarak spontan lokomotor aktivitenin belirlenmesi<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

Sosyal Kaçınma Niceleme için Basit Testi<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

Davranış Tahliller için Boyama Böcekler: anestezi ve Çiftleşme Davranışı<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

Sosyal Space Etkileyen Koşullar<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

için yiyecek arama yolu uzunluğu Protokolü<em&gt; Drosophila melanogaster</em&gt; Larva

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

Kılcal Besleyici Testi Gıda Alımını içinde Tedbirler<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

Drosophila melanogaster Larval ve yetişkin beyin metabolik Analizi

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

Drosophila melanogaster'de Akson Ölümü Sırasında Aksonların ve Sinapslarının Morfolojik ve Fonksiyonel Değerlendirilmesi

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