Name: an introduction to parasitic wasps of drosophila and the antiparasite immune response
Uploaded: 2012-05-07T12:00:00
Description: Watch this Scientific Journal Video about An Introduction to Parasitic Wasps of Drosophila and the Antiparasite Immune Response at JoVE.com

我々はTrichopria drosophilae、トランスジェニックハエの系統の教授トニーIP、および抗Spätzle抗体の教授カール·橋本教授はトッドSchlenkeに感謝しています。我々は、このプレゼンテーションへの貢献のためにラボの現在と過去のメンバーに感謝します。 NIH（S06 GM08168、RISE 41399から009、及びG12-RR03060）、USDA（NRI / USDA CSREES 2006から03817と2009-35302-05277）およびPSC-CUNYから：この作品は、以下の補助金によって支えられている。

実験のためのプロトコル全体は、4つのステップ（ 図9）に分割されています。ハエの幼虫（1）の培養ハチ（2）感染の設定と解剖のために動物を調製する工程;（3）ホスト/パラサイトの構造を分離し、固定（4）免疫組織を分析する。 1。 ショウジョウバエの幼虫の培養スズメバチスズメバチの文化の維持は、慎重な計画が必要です?...

<ol>
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K., Hartenstein, V., Banerjee, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+hematopoietic+stem+cell+and+its+niche:+a+comparative+view.">The hematopoietic stem cell and its niche: a comparative view.</a> Genes Dev. 21, 3044-3060 (2007).</li><li>Lemaitre, B., Hoffmann, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+host+defense+of+Drosophila+melanogaster.">The host defense of Drosophila melanogaster.</a> Annu. Rev. Immunol. 25, 697-743 (2007).</li><li>Schlegel, A., Stainier, D. 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A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+hemocyte-specific+enhancer-reporter+transgenes+for+the+analysis+of+hematopoiesis+in+Drosophila.">New hemocyte-specific enhancer-reporter transgenes for the analysis of hematopoiesis in Drosophila.</a> Genesis. 47, 771-774 (2009).</li><li>Tokusumi, T., Sorrentino, R. P., Russell, M., Ferrarese, R., Govind, S., Schulz, R. 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T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hemolymph-dependent+and+-independent+responses+in+Drosophila+immune+tissue.">Hemolymph-dependent and -independent responses in Drosophila immune tissue.</a> J. Cell Biochem. 92, 849-863 (2004).</li><li>Melk, J. P., Govind, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Developmental+analysis+of+Ganaspis+xanthopoda,+a+larval+parasitoid+of+Drosophila+melanogaster.">Developmental analysis of Ganaspis xanthopoda, a larval parasitoid of Drosophila melanogaster.</a> J. Exp. Biol. 202, 1885-1896 (1999).</li></ol>

ショウジョウバエの寄生蜂への関心が全ゲノムを解読するための分子技術は効率的かつ低コストになると急増している。しかし、彼らの非常によく研究されたホストへの相対的な、スズメバチの生物学の多くの魅惑的な側面が不明瞭なままである。これらは、宿主範囲、免疫抑制、過寄生、および動作に関連する問題が含まれています。このプレゼンテーションの焦点は、ハエの免疫...

<table border="1"><tbody><tr><td> 材料 </td><td> タイプ </td><td> 会社 </td><td> カタログ番号 </td></tr><tr><td> 昆虫培養維持のための材料 </td><td></td><td></td><td></td></tr><tr><td>酵母</td><td>アクティブドライ</td><td>フィッシャー·サイエンティフィック</td><td> S802453 </td></tr><tr><td>食べ物を飛ぶ</td><td>コー​​ンミール、砂糖</td><td></td><td>標準的なレシピ</td></tr><tr><td>ハニー</td><td>クローバー</td><td>オランダのゴールド</td><td></td></tr><tr><td>バイアル</td><td>ポリプロピレンシェルバイアル（ナロー） </td><td>フィッシャー·サイエンティフィック</td><td> AS514 </td></tr><tr><td>バイアルの閉鎖</td><td>綿栓</td><td>フィッシャー·サイエンティフィック</td><td> AS212 </td></tr><tr><td>バイアルの閉鎖</td><td>バズプラグ</td><tD&gt;ジェネシー科学</td><td> AS273 </td></tr><tr><td>冷蔵インキュベーター</td><td>精度815 </td><td>サーモフィッシャーサイエンティフィック</td><td> 3721 </td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td> サンプル調製のための材料 </td><td></td><td></td><td></td></tr><tr><td> CO 2タンク</td><td>骨乾燥したグレード</td><td> TWスミス</td><td> UN1013 </td></tr><tr><td>へら</td><td>マイクロスパチュラ（14センチ） </td><td>フィッシャー·サイエンティフィック</td><td> 21-401-15 </td></tr><tr><td>パイレックススポット試験板</td><td> 9よく解剖板85 mm x 100 mm幅</td><td>トーマス科学</td><td> 7812G17 </td></tr><tr><td>パスツールピペット</td><td>ソーダ石灰</td><td> J＆Hベルジェ</td><td> 71-5200-05 </td></tr><tr><td>鉗子</td><td>様式第5位</td><td>シグマ</td><td> T-4662 </td></tr><tr><td>エタノール</td><td> 190プルーフのUSP </td><td>フィッシャー·サイエンティフィック</td><td> 04-355-221 </td></tr><tr><td>ホルムアルデヒド</td><td> 37パーセント（w / w）の</td><td>フィッシャー·サイエンティフィック</td><td> F79-1 </td></tr><tr><td> Cy3標識二次抗体AffiniPureロバ抗ウサギIgG（H + L） </td><td>午前1時50分励起546 nmで、発光565 nmの</td><td>ジャクソンイムノリサーチ·ラボラトリーズ株式会社</td><td> 711-165-152 </td></tr><tr><td>退色（N-没食子酸プロピル） </td><td> 1X PBS、50％グリセロール溶液で4μg/ mlの</td><td> MPのバイオメディカル</td><td> 10274790 </td></tr><tr><td>グリセロール</td><td></td><td>フィッシャー·サイエンティフィック</td><td> G33-1 </td></tr><tr><td>ヘキスト33258 </td><td> 0.2μg/ mlの励起352 nmで、発光461 nmの</td><td>分子プローブ</td><td> H-1398 </td></tr><tr><td>ローダミンファロイジン</td><td> 200単位/ ml（6.6μM）励起波長540nm、発光565nmで</td><td>分子プローブ</td><td> R415 </td></tr><tr><td>のAlexa Fluor®488ファロイジン</td><td> 300単位/ mlの励起495nmの、発光518 nmの</td><td>分子プローブ</td><td> A12379 </td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td> 使い捨て </td><td></td><td></td><td></td></tr><tr><td>洗浄瓶</td><td> Fisherbrand </td><td>フィッシャー·サイエンティフィック</td><td> 03から409-22A </td></tr><tr><td>キムワイプ</td><td>キンバリークラーク</td><td>フィッシャー·サイエンティフィック</td><td> 06-666A </td></tr><tr><td>ペーパータオル</td><td> 1プライC-フォールド</td><td>クイル</td><td> 901-7CFTB2400 </td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td> 顕微鏡検査 </td><td> ＆回り、湧き出る！ </td><td></td><td></td></tr><tr><td>ライカ実体顕微鏡</td><td> MZFLIII </td><td>帝国イメージングシステムズ株式会社</td><td> 10446208 </td></tr><tr><td>ツァイス実体顕微鏡</td><td> STEMIの1000または2000-C </td><td>カールツァイス</td><td> 000000-1006-126 </td></tr><tr><td>光源 - LED </td><td>グースネック照明</td><td>フィッシャー·サイエンティフィック</td><td> 12563501 </td></tr><tr><td>ステージ</td><td>プレートと透過光ボックス</td><td>カールツァイス</td><td> 455137000 </td></tr><tr><td>ツァイスレーザー共焦点顕微鏡をスキャンする</td><td> LSM 510 </td><td>カールツァイス</td><td></td></tr><tr><td>ツァイスの複合顕微鏡</td><td>直立Axioplan 2 </td><td>カールツァイス</td><td></td></tr></tbody></table><table border="1"><tbody><tr><td> スズメバチの系統 </td><td> ひずみを飛ばすの</td></tr><tr><td> Leptopilina victoriae 16 </td><td> YW </td></tr><tr><td> Leptopilina boulardi 17 1 </td><td> UAS-GFP-背 17 </td></tr><tr><td> Leptopilina heterotoma 2 </td><td> SerpentHemoGal4 13 </td></tr><tr><td> Leptopilina heterotoma 14 1 </td><td> MSNF9-moCherry 14 </td></tr><tr><td> Trichopria drosophilae </td><td> MSNF-GFP 15 </td></tr><tr><td> Ganaspis xanthopoda 18 </td><td> YW蛇-GAL4 UAS GFP-Dorsal/Basc 4 </td></tr><tr><td></td><td> YW。Drosomycin-GFP/CyO Y + 12 </td></tr></tbody></table>

an introduction to parasitic wasps of drosophila and the antiparasite immune response

Most known parasitoid wasp species attack the larval or pupal stages of Drosophila. While Trichopria drosophilae infect the pupal stages of the host (Fig. 1A-C), females of the genus Leptopilina (Fig. 1D, 1F, 1G) and Ganaspis (Fig. 1E) attack the larval stages. We use these parasites to study the molecular basis of a biological arms race. Parasitic wasps have tremendous value as biocontrol agents. Most of them carry virulence and other factors that modify host physiology and immunity. Analysis of Drosophila wasps is providing insights into how species-specific interactions shape the genetic structures of natural communities. These studies also serve as a model for understanding the hosts' immune physiology and how coordinated immune reactions are thwarted by this class of parasites.
The larval/pupal cuticle serves as the first line of defense. The wasp ovipositor is a sharp needle-like structure that efficiently delivers eggs into the host hemocoel. Oviposition is followed by a wound healing reaction at the cuticle (Fig. 1C, arrowheads). Some wasps can insert two or more eggs into the same host, although the development of only one egg succeeds. Supernumerary eggs or developing larvae are eliminated by a process that is not yet understood. These wasps are therefore referred to as solitary parasitoids.
Depending on the fly strain and the wasp species, the wasp egg has one of two fates. It is either encapsulated, so that its development is blocked (host emerges; Fig. 2 left); or the wasp egg hatches, develops, molts, and grows into an adult (wasp emerges; Fig. 2 right). L. heterotoma is one of the best-studied species of Drosophila parasitic wasps. It is a "generalist," which means that it can utilize most Drosophila species as hosts1. L. heterotoma and L. victoriae are sister species and they produce virus-like particles that actively interfere with the encapsulation response2. Unlike L. heterotoma, L. boulardi is a specialist parasite and the range of Drosophila species it utilizes is relatively limited1. Strains of L. boulardi also produce virus-like particles3 although they differ significantly in their ability to succeed on D. melanogaster1. Some of these L. boulardi strains are difficult to grow on D. melanogaster1 as the fly host frequently succeeds in encapsulating their eggs. Thus, it is important to have the knowledge of both partners in specific experimental protocols.
In addition to barrier tissues (cuticle, gut and trachea), Drosophila larvae have systemic cellular and humoral immune responses that arise from functions of blood cells and the fat body, respectively. Oviposition by L. boulardi activates both immune arms1,4. Blood cells are found in circulation, in sessile populations under the segmented cuticle, and in the lymph gland. The lymph gland is a small hematopoietic organ on the dorsal side of the larva. Clusters of hematopoietic cells, called lobes, are arranged segmentally in pairs along the dorsal vessel that runs along the anterior-posterior axis of the animal (Fig. 3A). The fat body is a large multifunctional organ (Fig. 3B). It secretes antimicrobial peptides in response to microbial and metazoan infections.
Wasp infection activates immune signaling (Fig. 4)4. At the cellular level, it triggers division and differentiation of blood cells. In self defense, aggregates and capsules develop in the hemocoel of infected animals (Fig. 5)5,6. Activated blood cells migrate toward the wasp egg (or wasp larva) and begin to form a capsule around it (Fig. 5A-F). Some blood cells aggregate to form nodules (Fig. 5G-H). Careful analysis reveals that wasp infection induces the anterior-most lymph gland lobes to disperse at their peripheries (Fig. 6C, D).
We present representative data with Toll signal transduction pathway components Dorsal and Sp&auml;tzle (Figs. 4,5,7), and its target Drosomycin (Fig. 6), to illustrate how specific changes in the lymph gland and hemocoel can be studied after wasp infection. The dissection protocols described here also yield the wasp eggs (or developing stages of wasps) from the host hemolymph (Fig. 8).

Watch this Scientific Journal Video about An Introduction to Parasitic Wasps of Drosophila and the Antiparasite Immune Response at JoVE.com

An Introduction to Parasitic Wasps of Drosophila and the Antiparasite Immune Response

寄生蜂（寄生）ハチを含む多くの昆虫の天敵の主要なクラスを構成する<em&gt;キイロショウジョウバエ</em&gt;。我々は、これらの寄生虫を伝播するためのテクニックを紹介します<em&gt;ショウジョウバエ</em&gt;属。との免疫組織に与える影響を分析する方法を示しています<em&gt;ショウジョウバエ</em&gt;幼虫。

の寄生蜂の紹介<em&gt;ショウジョウバエ</em&gt;と抗寄生虫免疫応答

Most known parasitoid wasp species attack the larval or pupal stages of Drosophila. While Trichopria drosophilae infect the pupal stages of the host (Fig. ...

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Non-invasive Imaging of the Innate Immune Response in a Zebrafish Larval Model of Streptococcus iniae Infection

のゼブラフィッシュ幼生モデルにおける先天性免疫応答の非侵襲的イメージング<em&gt;ストレプトコッカスイニエ</em&gt;感染

The aquatic pathogen, Streptococcus iniae, is responsible for over 100 million dollars in annual losses for the aquaculture industry and is capable of ...

Inoculating Anopheles gambiae Mosquitoes with Beads to Induce and Measure the Melanization Immune Response

Inoculating Anopheles gambiae Mosquitoes with Beads to Induce and Measure the Melanization Immune Response

接種<em&gt;ハマダラカ</emメラニン免疫応答を誘導し、測定するためにビーズと&gt;蚊

The stimulation of immune responses is a common tool in invertebrate studies to examine the efficacy and the mechanisms of immunity. This stimulation is ...