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.

注：このプロトコルは、準備（セクション1から3）について説明し、2-Dの満腹アッセイの実行（第4節）、および分析（第4節）。次に、例として、ドーパミン作動性刺激を使用して、第5節では異常性欲を誘導するために2次元満腹アッセイと熱発生を起こす刺激を結合する方法を示しています。第6節では、2-Dの満腹アッセイの結果を検証する3つの方法について説明します。最後に、7節は、男?...

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動機づけ状態は、飽き飽きし維持、及び34を回収することができます。我々は、迅速かつ堅牢フライに交配ドライブのこれらの側面のすべてを測定する2次元満腹アッセイを提示します。このアッセイは、動機付け行動の分子や回路部品を研究するために、高度なハエの遺伝子操作を使用しての可能性を開きます。 満腹アッセイが正常に裁判所や交尾す?...

ショウジョウバエでの作業は、遺伝子1の性質を含め、多くの生物学的現象に深く、先駆的な洞察を提供している、胚発生2、概日リズム3、および神経系4、5、6の開発や配線の原則。動機は、おそらくため、これまで検討されているシステム上の制限により、これまであまりよく理解これらの現象よりも残っています。ハエにおける動機は主に脂肪沈着の明白な徴候を排除送り試合と外骨格あたりの無視できるほど小さい食物摂取に起因する多くの課題を提示し、飢餓、との関連で検討されています。したがって、ハエにモチベーションを研究するために使用されるシステムを拡張する必要があります。 我々は内交配ドライブの研究のための行動の枠組みを説明しますショウジョウバエ 。このシステムは、フライに神経遺伝ツールを利用するだけでなく、アクセシビリティ7、8、9、10、11、12、その性的二形回路8,13の推定コネクトーム。また、生来の14、15、16、17、18、19、20、21の多く及び22を学習し、23において、24感覚運動回路制御求愛は、貴重な機会を提供し、具体的に働いています動機が衝突する時に正確な回路ノードを検索します。我々は最近、フライに、人間のように、ドーパミンレベルが交配ドライブ25、26、27の中心である、と報告しました。私たちはここに25を説明するアッセイを使用して、この保存された現象の解析関連ドーパミン産生やフライにニューロンを受け、容易に詳細なmolecular-と回路レベルへの遺伝的アクセスを得ています。 私たちは、Zhang らで行動アッセイに追加します。 25我々は2次元（2-D）満腹アッセイを呼び出すビデオ得点、以前の方法に比べて重要な改善を可能にする新しいフラット行動アリーナ。その結果、新しいアッセイは、よりスケーラブルかつ定量化できる、とモチベーションに関与する遺伝子と神経細胞の遺伝子スクリーニングのために、したがって、より適しています。私たちは求愛のアッセイおよびneurogeと一緒に、この新しいアッセイを使用します磁操作は、測定し、ハエにおける交配ドライブを変更する方法を実証します。

<table border="0" cellpadding="0" cellspacing="0" width="1195">
	<tbody>
		<tr height="21">
			<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>
		</tr>
		<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>
		</tr>
		<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>
		</tr>
		<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>
		</tr>
	</tbody>
</table>

measuring and altering mating drive in male drosophila melanogaster

調査の何十年にもかかわらず、動機づけの状態の神経細胞および分子の塩基は、神秘的なままです。我々は最近、小説、還元主義、および男性キイロショウジョウバエ （ショウジョウバエ）、ここでは詳細をWEするための方法の交配ドライブを使用して動機の徹底的な調査のためのスケーラブルなシステムを開発しました。男性の交尾ドライブが繰り返さcopulationsの過程で生殖能力と一緒に減少し、〜3日間にわたって回復所見上の行動パラダイムセンター。このシステムでは、ハエで利用可能な強力な神経遺伝ツールは、遺伝的アクセシビリティと性的行動のために利用可能な推定上の配線図で収束します。この収束は、特定の動機づけの機能を持つ小型のニューロン集団の迅速な単離および尋問を可能にします。ここでは詳細男性のハエで求愛のモチベーションを測定し、変更するために使用される満腹アッセイの設計と実行を。これを使用して、アッセイは、我々は、低雄嵌合ドライブがドーパミン作動性ニューロンを刺激することによって克服することができることを示しています。満腹アッセイは、単純な手頃な価格、および遺伝的背景の影響に対してロバストです。私たちは、満腹アッセイは動機づけの状態の神経生物学に多くの新たな洞察を生成するために期待しています。

ショウジョウバエ嵌合ドライブを特徴づけるために、3日齢、WTカントン-S男性は、2-Dの満腹アッセイで試験しました。アッセイ（4.5時間）の間に、男性が4.8±0.3の平均を交尾回数（平均値、SEMの±標準誤差を意味します）。交配は、最初の2時間（78％）（ 図6A、6B）で、主に開始し、アッセイは（ 図6A、6B）を進むにつれて少なく頻繁に?...

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

この記事では、モチベーションを研究するために、ショウジョウバエの melanogaste rの雄交尾ドライブを使用する行動アッセイを記載します。この方法を使用して、研究者は、この動機の根底にある、分子遺伝学、および細胞メカニズムを解明するために、高度なフライ神経遺伝技術を利用することができます。

測定とマレの変更交尾ドライブ<em&gt;キイロショウジョウバエ</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

昆虫における単一感覚子録音キイロショウジョウバエとハマダラカ</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

で逃避行動のOptogenetic刺激<em&gt;キイロショウジョウバエ</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

における自発運動量の測定<em&gt;キイロショウジョウバエ</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

社会回避での定量化のための簡単​​なアッセイ<em&gt;キイロショウジョウバエ</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

行動アッセイの染色昆虫：麻酔の配偶行動<em&gt;ショウジョウバエ</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

で社会空間に影響を与える条件<em&gt;キイロショウジョウバエ</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

パスの長さのプロトコルを採餌<em&gt;キイロショウジョウバエ</em&gt;幼虫

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

キャピラリーフィーダーアッセイにおいて食物摂取を測定します<em&gt;キイロショウジョウバエ</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

キイロショウジョウバエ幼虫と大人の脳の代謝解析

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

ショウジョウバエメラノガスターにおける軸索死における軸索とそのシナプスの形態学的および機能的評価

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