Estamos muy agradecidos con el profesor Todd Schlenke Trichopria de mosca de la fruta, el profesor Tony Ip cepas de moscas transgénicas, y el Prof. Carl Hashimoto para el anti-Spätzle anticuerpos. Damos las gracias a los miembros actuales y anteriores del laboratorio por sus contribuciones a esta presentación. Este trabajo fue apoyado por las siguientes subvenciones: a partir de los NIH (S06 GM08168, RISE 41399-009, y G12 RR03060-), el USDA (NRI / USDA CSREES 2006-03817 y 2009-35302-05277) y PSC-CUNY.

<ol>
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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. 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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=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. 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No hay conflictos de interés declarado.

El interés por las avispas parásitas de Drosophila está surgiendo como las técnicas moleculares para descifrar genomas completos se vuelven eficientes y rentables. Sin embargo, en relación con sus excepcionalmente bien estudiados los ejércitos, muchos aspectos fascinantes de la biología avispas siguen siendo oscuros. Estos incluyen temas relacionados con la gama de huéspedes, la inmunosupresión, superparasitismo, y el comportamiento. El objetivo de esta presentación fue demostrar los efectos de la inf...

<table border="1"><tbody><tr><td> Materiales </td><td> Tipo </td><td> Empresa </td><td> Número de catálogo </td></tr><tr><td> Materiales para el mantenimiento de los cultivos de insectos </td><td></td><td></td><td></td></tr><tr><td> Levadura </td><td> Seca activa </td><td> Fisher Scientific </td><td> S802453 </td></tr><tr><td> Fly alimentos </td><td> La harina de maíz, el azúcar </td><td></td><td> Receta estándar </td></tr><tr><td> Miel </td><td> Trébol </td><td> Oro holandesa </td><td></td></tr><tr><td> Viales </td><td> Polipropileno shell viales (estrecho) </td><td> Fisher Scientific </td><td> AS514 </td></tr><tr><td> Cierres de los viales </td><td> Algodón enchufe </td><td> Fisher Scientific </td><td> AS212 </td></tr><tr><td> Cierres de los viales </td><td> Zumbido enchufe </td><td&gt; Genesee Científico </td><td> AS273 </td></tr><tr><td> Estufa refrigerada </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> Los materiales para la preparación de muestras </td><td></td><td></td><td></td></tr><tr><td> Cilindro de CO 2 </td><td> Grado de sequedad del hueso </td><td> TW Smith </td><td> UN1013 </td></tr><tr><td> Espátula </td><td> Espátula de micro (14 cm) </td><td> Fisher Scientific </td><td> 21-401-15 </td></tr><tr><td> Pyrex placas planas de prueba </td><td> 9-así placa de la disección de 85 mm x 100 mm </td><td> Thomas Scientific </td><td> 7812G17 </td></tr><tr><td> Pipetas Pasteur </td><td> La cal sodada </td><td> J &amp; H Berge </td><td> 71-5200-05 </td></tr><tr><td> Fórceps </td><td> Estilo # 5 </td><td> Sigma </td><td> T-4662 </td></tr><tr><td> Etanol </td><td> 190 prueba de la USP </td><td> Fisher Scientific </td><td> 04-355-221 </td></tr><tr><td> Formaldehído </td><td> 37% w / w </td><td> Fisher Scientific </td><td> F79-1 </td></tr><tr><td> Anticuerpo secundario Cy3 AffiniPure burro anti-conejo IgG (H + L) </td><td> 1:50 excitación de 546 nm; de emisión 565 nm </td><td> Jackson Immuno Research Laboratories, Inc. </td><td> 711-165-152 </td></tr><tr><td> Antifade (N-propil galato) </td><td> 4 mg / ml en glicerol al 50% en 1X PBS </td><td> MP Biomedicals </td><td> 10274790 </td></tr><tr><td> Glicerol </td><td></td><td> Fisher Scientific </td><td> G-33-1 </td></tr><tr><td> Hoechst 33258 </td><td> 0,2 g / ml de excitación 352 nm, emisión 461 nm </td><td> Molecular Probes </td><td> H-1398 </td></tr><tr><td> Rodamina phalloidin </td><td> 200 unidades / ml (6,6Excitación mM) 540 nm; de emisión 565 nm </td><td> Molecular Probes </td><td> R415 </td></tr><tr><td> Alexa Fluor 488 phalloidin </td><td> 300 unidades / ml de excitación de 495 nm, emisión 518 nm </td><td> Molecular Probes </td><td> A12379 </td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td> Desechables </td><td></td><td></td><td></td></tr><tr><td> Botella de lavado </td><td> Fisherbrand </td><td> Fisher Scientific </td><td> 03 a 409-22A </td></tr><tr><td> Kimwipes </td><td> Kimberly Clark </td><td> Fisher Scientific </td><td> 06-666A </td></tr><tr><td> Toallas de papel </td><td> 1 capa C-Fold </td><td> Pluma </td><td> 901-7CFTB2400 </td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td> Microscopía </td><td> Ynbsp; </td><td></td><td></td></tr><tr><td> Leica estereomicroscopio </td><td> MZFLIII </td><td> Imperio Imaging Systems, Inc. </td><td> 10446208 </td></tr><tr><td> Zeiss Estereomicroscopio </td><td> Stemi 1000 o 2000, C- </td><td> Carl Zeiss </td><td> 000000-1006-126 </td></tr><tr><td> Fuente de Luz - LED </td><td> Cuello de cisne iluminador </td><td> Fisher Scientific </td><td> 12563501 </td></tr><tr><td> Etapa </td><td> Caja de luz transmitida con placa </td><td> Carl Zeiss </td><td> 455137000 </td></tr><tr><td> Zeiss microscopio confocal de barrido láser </td><td> LSM 510 </td><td> Carl Zeiss </td><td></td></tr><tr><td> Microscopio compuesto Zeiss </td><td> Axioplan 2 en posición vertical </td><td> Carl Zeiss </td><td></td></tr></tbody></table><table border="1"><tbody><tr><td> Las cepas de avispa </td><td> Fly Cueles </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 drosófilas </td><td> Sólidos lácteos no grasos 15-GFP </td></tr><tr><td> Ganaspis xanthopoda 18 </td><td> YW-Serpiente Gal4 UAS GFP-Dorsal/Basc 4 </td></tr><tr><td></td><td> PA; 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

Parasitoide (parásitos) avispas constituyen una clase importante de enemigos naturales de muchos insectos, incluyendo<em&gt; Drosophila melanogaster</em&gt;. Vamos a introducir las técnicas para propagar estos parásitos en<em&gt; Drosophila</em&gt; Spp. y demostrar la forma de analizar sus efectos sobre los tejidos inmunes de<em&gt; Drosophila</em&gt; Larvas.

Una introducción a las avispas parasitarias de<em&gt; Drosophila</em&gt; Y la respuesta inmune antiparasitario

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 vistas.  The City College of New York, CUNY. Parasitoide (parásitos) avispas constituyen una clase importante de enemigos naturales de muchos insectos, incluyendo Drosophila melanogaster. Vamos a introducir las técnicas para propagar estos parásitos en Drosophila Spp. y demostrar la forma de analizar sus efectos sobre los tejidos inmunes de Drosophila Larvas.