Wir danken Prof. Todd Schlenke für Trichopria drosophilae, Prof. Tony Ip für transgene Fliegen-Stämmen, und Prof. Carl Hashimoto für anti-Spätzle-Antikörper. Wir bedanken uns bei gegenwärtigen und früheren Mitgliedern des Labors für ihre Beiträge zu dieser Präsentation. Vom NIH (GM08168 S06, RISE 41399-009 und G12-RR03060), USDA (NRI / USDA CSREES 2006-03817 und 2009-35302-05277) und PSC-CUNY: Diese Arbeit wurde von den folgenden Zuschüssen unterstützt.

<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=A+drosomycin-GFP+reporter+transgene+reveals+a+local+immune+response+in+Drosophila+that+is+not+dependent+on+the+Toll+pathway.">A drosomycin-GFP reporter transgene reveals a local immune response in Drosophila that is not dependent on the Toll pathway.</a> EMBO J. 17, 1217-1227 (1998).</li><li>Bruckner, K., Kockel, L., Duchek, P., Luque, C. M., Rorth, P., Perrimon, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+PDGF/VEGF+receptor+controls+blood+cell+survival+in+Drosophila.">The PDGF/VEGF receptor controls blood cell survival in Drosophila.</a> Dev. Cell. 7, 73-84 (2004).</li><li>Tokusumi, T., Shoue, D. A., Tokusumi, Y., Stoller, J. R., Schulz, R. 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Keine Interessenskonflikte erklärt.

Das Interesse an Schlupfwespen von Drosophila wird als molekulare Techniken, um ganze Genome entschlüsseln werden effiziente und kostengünstige sprunghaft zu. Jedoch im Verhältnis zu ihrem außergewöhnlich gut untersuchten Gastgeber, bleiben viele faszinierende Aspekte der Biologie Wespe im Dunkeln. Dazu gehören Fragen im Zusammenhang mit Reichweite, Immunsuppression, Superparasitismus und Verhalten zu hosten. Der Schwerpunkt dieser Präsentation war es, die Auswirkungen der Infektion auf der Fliege Immuns...

<table border="1"><tbody><tr><td> Materialien </td><td> Typ </td><td> Firma </td><td> Katalog-Nummer </td></tr><tr><td> Materialien für Insekten Kultur Wartung </td><td></td><td></td><td></td></tr><tr><td> Hefe </td><td> Aktive Trockenhefe </td><td> Fisher Scientific </td><td> S802453 </td></tr><tr><td> Fly Lebensmittel </td><td> Maismehl, Zucker </td><td></td><td> Standard-Rezept </td></tr><tr><td> Honig </td><td> Klee </td><td> Niederländischen Gold- </td><td></td></tr><tr><td> Vials </td><td> Sitzschale aus Polypropylen Ampullen (schmal) </td><td> Fisher Scientific </td><td> AS514 </td></tr><tr><td> Vial Schließungen </td><td> Wattestopfen </td><td> Fisher Scientific </td><td> AS212 </td></tr><tr><td> Vial Schließungen </td><td> Buzz-Stecker </td><td&gt; Genesee Scientific </td><td> AS273 </td></tr><tr><td> Kühlbrutschrank </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> Materialien für die Probenvorbereitung </td><td></td><td></td><td></td></tr><tr><td> CO 2-Tank </td><td> Knochentrocken Klasse </td><td> TW Smith </td><td> UN1013 </td></tr><tr><td> Spachtel </td><td> Micro Spatel (14 cm) </td><td> Fisher Scientific </td><td> 21-401-15 </td></tr><tr><td> Pyrex Spot Testplatten </td><td> 9-gut sezieren Platte 85 mm x 100 mm </td><td> Thomas Scientific </td><td> 7812G17 </td></tr><tr><td> Pasteurpipetten </td><td> Atemkalk </td><td> J &amp; H Berge </td><td> 71-5200-05 </td></tr><tr><td> Zange </td><td> Style # 5 </td><td> Sigma </td><td> T-4662 </td></tr><tr><td> Ethanol </td><td> 190 Beweis USP </td><td> Fisher Scientific </td><td> 04-355-221 </td></tr><tr><td> Formaldehyd </td><td> 37% w / w </td><td> Fisher Scientific </td><td> F79-1 </td></tr><tr><td> Sekundärer Antikörper Cy3 AffiniPure Esel-anti-Kaninchen-IgG (H + L) </td><td> 01.50 Excitation 546 nm; Emission 565 nm </td><td> Jackson Immuno Research Laboratories, Inc. </td><td> 711-165-152 </td></tr><tr><td> Antifade (N-Propylgallat) </td><td> 4 ug / ml in 50% Glycerin in 1X PBS </td><td> MP Biomedicals </td><td> 10274790 </td></tr><tr><td> Glycerin </td><td></td><td> Fisher Scientific </td><td> G33-1 </td></tr><tr><td> Hoechst 33258 </td><td> 0,2 ug / ml Excitation 352 nm; Emission 461 nm </td><td> Molecular Probes </td><td> H-1398 </td></tr><tr><td> Rhodamin-Phalloidin </td><td> 200 Einheiten / ml (6,6uM) Excitation 540 nm; Emission 565 nm </td><td> Molecular Probes </td><td> R415 </td></tr><tr><td> Alexa Fluor 488 Phalloidin </td><td> 300 Einheiten / ml Excitation 495 nm; Emission 518 nm </td><td> Molecular Probes </td><td> A12379 </td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td> Verbrauchsmaterial </td><td></td><td></td><td></td></tr><tr><td> Waschflasche </td><td> Fisherbrand </td><td> Fisher Scientific </td><td> 03 bis 409-22A </td></tr><tr><td> Kimwipes </td><td> Kimberly Clark </td><td> Fisher Scientific </td><td> 06-666A </td></tr><tr><td> Papierhandtuchspender </td><td> 1-lagig C-Falz </td><td> Federkiel </td><td> 901-7CFTB2400 </td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td> Mikroskopie </td><td> &amp;nbsp; </td><td></td><td></td></tr><tr><td> Leica Stereomikroskop </td><td> MZFLIII </td><td> Reich Imaging Systems, Inc. </td><td> 10446208 </td></tr><tr><td> Zeiss Stereomikroskop </td><td> Stemi 1000 oder 2000-C </td><td> Carl Zeiss </td><td> 000000-1006-126 </td></tr><tr><td> Light Source - LED </td><td> Schwanenhals-Hilfslicht </td><td> Fisher Scientific </td><td> 12563501 </td></tr><tr><td> Bühne </td><td> Durchlicht-Box mit Platte </td><td> Carl Zeiss </td><td> 455137000 </td></tr><tr><td> Zeiss Laser-Scanning-konfokalen Mikroskop </td><td> LSM 510 </td><td> Carl Zeiss </td><td></td></tr><tr><td> Zeiss zusammengesetzte Mikroskop </td><td> Axioplan 2 aufrecht </td><td> Carl Zeiss </td><td></td></tr></tbody></table><table border="1"><tbody><tr><td> Wespen-Stämme </td><td> Fly abseihens </td></tr><tr><td> Leptopilina victoriae 16 </td><td> yw </td></tr><tr><td> Leptopilina boulardi 17 1 </td><td> UAS-GFP-Dorsal 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 UAS-Gal4 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

Parasitoiden (parasitäre) Wespen stellen eine große Klasse von natürlichen Feinde vieler Insekten einschließlich<em&gt; Drosophila melanogaster</em&gt;. Wir stellen Ihnen die Techniken, diese Parasiten in propagieren<em&gt; Drosophila</em&gt; Spp. und demonstrieren, wie man ihre Wirkung auf das Immunsystem Gewebe analysieren<em&gt; Drosophila</em&gt; Larven.

Eine Einführung in die parasitischen Wespen<em&gt; Drosophila</em&gt; Und die antiparasitäre 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. ...

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

18.4K Aufrufe.  The City College of New York, CUNY. Parasitoiden (parasitäre) Wespen stellen eine große Klasse von natürlichen Feinde vieler Insekten einschließlich Drosophila melanogaster. Wir stellen Ihnen die Techniken, diese Parasiten in propagieren Drosophila Spp. und demonstrieren, wie man ihre Wirkung auf das Immunsystem Gewebe analysieren Drosophila Larven.