Siamo grati al professor Todd Schlenke per Trichopria drosophilae, Prof. Tony Ip per i ceppi transgenici mosca, e il Prof. Carl Hashimoto per gli anticorpi anti-Spätzle. Ringraziamo i membri presenti e passati del laboratorio per il loro contributo a questa presentazione. Questo lavoro è stato supportato da i seguenti aiuti: dal NIH (S06 GM08168, RISE 41399-009 e G12-RR03060), USDA (NRI / USDA CSREES 2006-03817 e 2009-35302-05277) e PSC-CUNY.

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L&#39;interesse per le vespe parassite di Drosophila è in aumento come tecniche molecolari per decodificare genomi interi diventare efficiente e conveniente. Tuttavia, rispetto ai loro eccezionalmente ben studiati padroni di casa, molti aspetti affascinanti della biologia vespa restano oscuri. Questi includono le questioni relative ad ospitare gamma, soppressione immunitaria, superparasitism, e il comportamento. L&#39;obiettivo di questa presentazione è stato quello di dimostrare gli effetti di contagio sui t...

<table border="1"><tbody><tr><td> Materiale </td><td> Tipo </td><td> Azienda </td><td> Numero di catalogo </td></tr><tr><td> Materiali per la manutenzione della cultura insetti </td><td></td><td></td><td></td></tr><tr><td> Lievito </td><td> Secco attivo </td><td> Fisher Scientific </td><td> S802453 </td></tr><tr><td> Vola alimentare </td><td> Farina di mais, zucchero </td><td></td><td> Ricetta standard </td></tr><tr><td> Miele </td><td> Trifoglio </td><td> Olandese Oro </td><td></td></tr><tr><td> Fiale </td><td> Polipropilene shell vial (stretto) </td><td> Fisher Scientific </td><td> AS514 </td></tr><tr><td> Vial chiusura </td><td> Cotton spina </td><td> Fisher Scientific </td><td> AS212 </td></tr><tr><td> Vial chiusura </td><td> Buzz spina </td><td&gt; Genesee Scientific </td><td> AS273 </td></tr><tr><td> Incubatore refrigerato </td><td> Precision 815 </td><td> Thermo Scientific </td><td> 3721 </td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td> Materiali per la preparazione del campione </td><td></td><td></td><td></td></tr><tr><td> CO 2 serbatoio </td><td> Bone grade a secco </td><td> TW Smith </td><td> UN1013 </td></tr><tr><td> Spatola </td><td> Spatola Micro (14 cm) </td><td> Fisher Scientific </td><td> 21-401-15 </td></tr><tr><td> Pyrex piastre di prova in loco </td><td> 9-ben dissezione piastra 85 mm x 100 mm </td><td> Thomas Scientific </td><td> 7812G17 </td></tr><tr><td> Pipette Pasteur </td><td> Soda lime </td><td> J &amp; H Berge </td><td> 71-5200-05 </td></tr><tr><td> Forcipe </td><td> Style # 5 </td><td> Sigma </td><td> T-4662 </td></tr><tr><td> Etanolo </td><td> 190 prova USP </td><td> Fisher Scientific </td><td> 04-355-221 </td></tr><tr><td> Formaldeide </td><td> 37% w / w </td><td> Fisher Scientific </td><td> F79-1 </td></tr><tr><td> Anticorpo secondario Cy3 AffiniPure asino anti-IgG di coniglio (H + L) </td><td> 01:50 eccitazione 546 nm, emissione 565 nm </td><td> Jackson Immuno Research Laboratories, Inc. </td><td> 711-165-152 </td></tr><tr><td> Antifade (N-propil gallato) </td><td> 4 microgrammi / ml in 50% glicerolo in 1X PBS </td><td> MP Biomedicals </td><td> 10274790 </td></tr><tr><td> Glicerina </td><td></td><td> Fisher Scientific </td><td> G33-1 </td></tr><tr><td> Hoechst 33258 </td><td> 0,2 ug / ml di eccitazione 352 nm, emissione 461 nm </td><td> Molecular Probes </td><td> H-1398 </td></tr><tr><td> Rhodamine phalloidin </td><td> 200 unità / ml (6,6pM) di eccitazione 540 nm, emissione 565 nm </td><td> Molecular Probes </td><td> R415 </td></tr><tr><td> Alexa Fluor 488 phalloidin </td><td> 300 unità / ml di eccitazione 495 nm, emissione 518 nm </td><td> Molecular Probes </td><td> A12379 </td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td> Disposables </td><td></td><td></td><td></td></tr><tr><td> Bottiglia di lavaggio </td><td> FISHERBRAND </td><td> Fisher Scientific </td><td> 03-409-22A </td></tr><tr><td> Kimwipes </td><td> Kimberly Clark </td><td> Fisher Scientific </td><td> 06-666A </td></tr><tr><td> Tovagliolo di carta </td><td> 1 strato C-Fold </td><td> Penna </td><td> 901-7CFTB2400 </td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td> Microscopia </td><td> &amp;nbsp; </td><td></td><td></td></tr><tr><td> Stereomicroscopio Leica </td><td> MZFLIII </td><td> Empire Imaging Systems, Inc. </td><td> 10446208 </td></tr><tr><td> Stereomicroscopio Zeiss </td><td> Stemi 1000 o 2000-C </td><td> Carl Zeiss </td><td> 000000-1006-126 </td></tr><tr><td> Sorgente luminosa - LED </td><td> Gooseneck illuminatore </td><td> Fisher Scientific </td><td> 12563501 </td></tr><tr><td> Fase </td><td> Trasmesso light box con piastra </td><td> Carl Zeiss </td><td> 455137000 </td></tr><tr><td> Zeiss microscopio confocale a scansione laser </td><td> LSM 510 </td><td> Carl Zeiss </td><td></td></tr><tr><td> Zeiss microscopio composto </td><td> Axioplan 2 montante </td><td> Carl Zeiss </td><td></td></tr></tbody></table><table border="1"><tbody><tr><td> Wasp Ceppi </td><td> Vola Strains </td></tr><tr><td> Leptopilina victoriae 16 </td><td> yw </td></tr><tr><td> Leptopilina Boulardi 17 1 </td><td> UAS-GFP-dorsale 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 Serpent-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).

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

Parassitoide (parassita) vespe costituiscono una classe importante di nemici naturali di insetti tra cui<em&gt; Drosophila melanogaster</em&gt;. Introdurremo le tecniche per diffondere questi parassiti in<em&gt; Drosophila</em&gt; Spp. e dimostrare come analizzare i loro effetti sui tessuti del sistema immunitario di<em&gt; Drosophila</em&gt; Larve.

An Introduction to vespe parassite di<em&gt; Drosophila</em&gt; E la risposta immunitaria antiparassitari

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

18.4K Views.  The City College of New York, CUNY. Parassitoide (parassita) vespe costituiscono una classe importante di nemici naturali di insetti tra cui Drosophila melanogaster. Introdurremo le tecniche per diffondere questi parassiti in Drosophila Spp. e dimostrare come analizzare i loro effetti sui tessuti del sistema immunitario di Drosophila Larve.