Nous sommes reconnaissants au professeur Todd Schlenke pour Trichopria drosophilae, le professeur Tony Ip pour les souches de mouches transgéniques, et le professeur Carl Hashimoto anti-Spätzle anticorps. Nous tenons à remercier les membres actuels et passés du laboratoire pour leurs contributions à cette présentation. Ce travail a été soutenu par les subventions suivantes: des NIH (S06 GM08168, RISE 41399-009, et G12-RR03060), USDA (NRI / USDA CSREES 2006-03817 et 2009-35302-05277) et de la CFP-CUNY.

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P., Carton, Y., 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=Cellular+immune+response+to+parasite+infection+in+the+Drosophila+lymph+gland+is+developmentally+regulated.">Cellular immune response to parasite infection in the Drosophila lymph gland is developmentally regulated.</a> Dev. Biol. , 243-265 (2002).</li><li>Sorrentino, R. P., 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=Genetic+analysis+of+contributions+of+dorsal+group+and+JAK-Stat92E+pathway+genes+to+larval+hemocyte+concentration+and+the+egg+encapsulation+response+in+Drosophila.">Genetic analysis of contributions of dorsal group and JAK-Stat92E pathway genes to larval hemocyte concentration and the egg encapsulation response in Drosophila.</a> Genetics. 166, 1343-1356 (2004).</li><li>Jung, S. H., Evans, C. J., Uemura, C., 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+Drosophila+lymph+gland+as+a+developmental+model+of+hematopoiesis.">The Drosophila lymph gland as a developmental model of hematopoiesis.</a> Development. 132, 2521-2533 (2005).</li><li>Krzemien, J., Crozatier, M., Vincent, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ontogeny+of+the+Drosophila+larval+hematopoietic+organ,+hemocyte+homeostasis+and+the+dedicated+cellular+immune+response+to+parasitism.">Ontogeny of the Drosophila larval hematopoietic organ, hemocyte homeostasis and the dedicated cellular immune response to parasitism.</a> Int. J. Dev. Biol. 54, 1117-1125 (2010).</li><li>Martinez-Agosto, J. A., Mikkola, H. 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. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lessons+from+"lower"+organisms:+what+worms,+flies,+and+zebrafish+can+teach+us+about+human+energy+metabolism.">Lessons from "lower" organisms: what worms, flies, and zebrafish can teach us about human energy metabolism.</a> PLoS Genet. 3, e199 (2007).</li><li>Ferrandon, D., Jung, A. C., Criqui, M., Lemaitre, B., Uttenweiler-Joseph, S., Michaut, L., Reichhart, J., Hoffmann, J. 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. 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. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+a+lamellocyte+transcriptional+enhancer+located+within+the+misshapen+gene+of+Drosophila+melanogaster.">Characterization of a lamellocyte transcriptional enhancer located within the misshapen gene of Drosophila melanogaster.</a> PLoS One. 4, e6429 (2009).</li><li>Morales, J., Chiu, H., Oo, T., Plaza, R., Hoskins, S., 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=Biogenesis,+structure,+and+immune-suppressive+effects+of+virus-like+particles+of+a+Drosophila+parasitoid,+Leptopilina+victoriae.">Biogenesis, structure, and immune-suppressive effects of virus-like particles of a Drosophila parasitoid, Leptopilina victoriae.</a> J. Insect Physiol. 51, 181-195 (2005).</li><li>Bettencourt, R., Asha, H., Dearolf, C., Ip, Y. 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>

Pas de conflits d&#39;intérêt déclarés.

L&#39;intérêt pour les guêpes parasites de la drosophile est en pleine progression que les techniques moléculaires pour décoder les génomes entiers devenir efficace et rentable. Toutefois, par rapport à leurs exceptionnellement bien étudiés hôtes, de nombreux aspects fascinants de la biologie de guêpe restent obscures. Il s&#39;agit notamment de questions liées à la gamme d&#39;hôtes, la suppression immunitaire, superparasitisme, et le comportement. L&#39;objectif de cette présentation était de ...

<table border="1"><tbody><tr><td> Matériels </td><td> Type </td><td> Entreprise </td><td> Le numéro de catalogue </td></tr><tr><td> Matériel pour entretien de la culture des insectes </td><td></td><td></td><td></td></tr><tr><td> Levure </td><td> Sèche active </td><td> Fisher Scientific </td><td> S802453 </td></tr><tr><td> Fly alimentaire </td><td> Farine de maïs, le sucre </td><td></td><td> Recette standard </td></tr><tr><td> Miel </td><td> Trèfle </td><td> Néerlandaise d&#39;or </td><td></td></tr><tr><td> Flacons </td><td> Polypropylène flacons shell (étroit) </td><td> Fisher Scientific </td><td> AS514 </td></tr><tr><td> Fermetures Vial </td><td> Tampon de coton </td><td> Fisher Scientific </td><td> AS212 </td></tr><tr><td> Fermetures Vial </td><td> Buzz fiche </td><td&gt; Genesee scientifique </td><td> AS273 </td></tr><tr><td> Incubateur réfrigéré </td><td> Précision 815 </td><td> Thermo Scientific </td><td> 3721 </td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td> Matériaux pour la préparation des échantillons </td><td></td><td></td><td></td></tr><tr><td> Réservoir de CO 2 </td><td> Os sec de qualité </td><td> TW Smith </td><td> UN1013 </td></tr><tr><td> Spatule </td><td> Spatule Micro (14 cm) </td><td> Fisher Scientific </td><td> 21-401-15 </td></tr><tr><td> Plaques d&#39;essai en Pyrex place </td><td> 9-bien la plaque de dissection 85 mm X 100 mm </td><td> Thomas scientifique </td><td> 7812G17 </td></tr><tr><td> Pasteur Pipettes </td><td> La chaux sodée </td><td> J &amp; H Berge </td><td> 71-5200-05 </td></tr><tr><td> Forceps </td><td> Style # 5 </td><td> Sigma </td><td> T-4662 </td></tr><tr><td> L&#39;éthanol </td><td> 190 USP preuve </td><td> Fisher Scientific </td><td> 04-355-221 </td></tr><tr><td> Formaldéhyde </td><td> 37% p / p </td><td> Fisher Scientific </td><td> F79-1 </td></tr><tr><td> Anticorps secondaire Cy3 AffiniPure d&#39;âne anti-IgG de lapin (H + L) </td><td> 01:50 excitation 546 nm; émission 565 nm </td><td> Jackson Immuno Research Laboratories, Inc </td><td> 711-165-152 </td></tr><tr><td> Antifade (N-gallate de propyle) </td><td> 4 ug / ml dans du glycérol 50% dans du PBS 1X </td><td> MP Biomedicals </td><td> 10274790 </td></tr><tr><td> Glycérol </td><td></td><td> Fisher Scientific </td><td> G33-1 </td></tr><tr><td> Hoechst 33258 </td><td> 0,2 pg / ml d&#39;excitation 352 nm; émission 461 nm </td><td> Molecular Probes </td><td> H-1398 </td></tr><tr><td> Rhodamine phalloïdine </td><td> 200 unités / ml (6,6uM) d&#39;excitation 540 nm; émission 565 nm </td><td> Molecular Probes </td><td> R415 </td></tr><tr><td> Alexa Fluor 488 phalloïdine </td><td> 300 unités / ml d&#39;excitation 495 nm; émission 518 nm </td><td> Molecular Probes </td><td> A12379 </td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td> Jetables </td><td></td><td></td><td></td></tr><tr><td> Pissette </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> Essuie-tout </td><td> 1 pli pliage en C </td><td> Penne </td><td> 901-7CFTB2400 </td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td> Microscopie </td><td> &amp;nbsp; </td><td></td><td></td></tr><tr><td> Stéréomicroscope Leica </td><td> MZFLIII </td><td> Empire Imaging Systems, Inc </td><td> 10446208 </td></tr><tr><td> Stéréomicroscope Zeiss </td><td> Stemi 1000 ou 2000 à C </td><td> Carl Zeiss </td><td> 000000-1006-126 </td></tr><tr><td> Source lumineuse - LED </td><td> Illuminateur col de cygne </td><td> Fisher Scientific </td><td> 12563501 </td></tr><tr><td> Étape </td><td> Transmis boîte à lumière avec plaque </td><td> Carl Zeiss </td><td> 455137000 </td></tr><tr><td> Zeiss microscope confocal à balayage laser </td><td> LSM 510 </td><td> Carl Zeiss </td><td></td></tr><tr><td> Microscope composé Zeiss </td><td> Axioplan 2 debout </td><td> Carl Zeiss </td><td></td></tr></tbody></table><table border="1"><tbody><tr><td> Les souches Wasp </td><td> Fly Filtrers </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> SS-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

Parasitoïde (parasite) guêpes constituent une classe importante des ennemis naturels des insectes, y compris de nombreux<em&gt; Drosophila melanogaster</em&gt;. Nous allons introduire les techniques de propager ces parasites dans les<em&gt; Drosophila</em&gt; Spp. et de démontrer la façon d&#39;analyser leurs effets sur les tissus immunitaires de<em&gt; Drosophila</em&gt; Larves.

Une introduction à guêpes parasites<em&gt; Drosophila</em&gt; Et la réponse immunitaire antiparasitaire

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 vues.  The City College of New York, CUNY. Parasitoïde (parasite) guêpes constituent une classe importante des ennemis naturels des insectes, y compris de nombreux Drosophila melanogaster. Nous allons introduire les techniques de propager ces parasites dans les Drosophila Spp. et de démontrer la façon d'analyser leurs effets sur les tissus immunitaires de Drosophila Larves.