Name: an introduction to parasitic wasps of drosophila and the antiparasite immune response
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אנו מודים לפרופ &#39;טוד Schlenke עבור Trichopria drosophilae, פרופ&#39; טוני Ip עבור זני זבובים מהונדס, ופרופ &#39;קרל השימוטו עבור אנטי Spätzle נוגדנים. אנו מודים לחברי ההווה ושל העבר הקרוב של המעבדה על תרומתם במצגת זו. עבודה זו נתמכה על ידי מענקים הבאים: מ-NIH (S06 GM08168, עלייתו 41399-009, G12 ו-RR03060), משרד החקלאות האמריקאי (NRI / משרד החקלאות CSREES 2006-03817 ו 2009-35302-05277) ו PSC-CUNY.

פרוטוקול הניסוי כולו מחולק לארבעה שלבים (איור 9). (1) culturing צרעות על זחלי זבוב, (2) הגדרת זיהומים והכנת חיות לנתיחה, (3) בידוד ותיקון המארח / טפיל מבנים: (4) ניתוח רקמות חיסוניות. 1. Culturing צרעות על הזחלים תסיסנית <p class="jove_content" ...

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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>

עניין צרעות טפיליות של תסיסנית גועשת כמו שיטות מולקולריות כדי לפענח גנומים שלמים להיות יעיל וחסכוני. עם זאת, ביחס במיוחד שלהם היטב למד המארחים, היבטים מרתקים רבים של הביולוגיה צרעה להישאר אלמוני. אלה כוללים נושאים הקשורים לארח מגוון, דיכוי המערכת החיסונית, superpara...

<table border="1" style=";text-align:right;direction:rtl"><tbody><tr><td> חומרים </td><td> סוג </td><td> חברה </td><td> מספר קטלוגי </td></tr><tr><td> חומרי תחזוקה התרבות חרקים </td><td></td><td></td><td></td></tr><tr><td> שמרים </td><td> פעיל יבש </td><td> פישר סיינטיפיק </td><td> S802453 </td></tr><tr><td> טוס מזון </td><td> קמח תירס, סוכר </td><td></td><td> מתכון רגיל </td></tr><tr><td> דבש </td><td> תלתן </td><td> ההולנדי זהב </td><td></td></tr><tr><td> בקבוקונים </td><td> פוליפרופילן פגז בקבוקונים (צר) </td><td> פישר סיינטיפיק </td><td> AS514 </td></tr><tr><td> הבקבוקון סגרים </td><td> קוטון תקע </td><td> פישר סיינטיפיק </td><td> AS212 </td></tr><tr><td> הבקבוקון סגרים </td><td> באז התוספת </td><tד&gt; Genesee מדעי </td><td> AS273 </td></tr><tr><td> בקירור החממה </td><td> דיוק 815 </td><td> Thermo Scientific </td><td> 3721 </td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td> חומרים עבור הכנת המדגם </td><td></td><td></td><td></td></tr><tr><td> CO 2 מיכל </td><td> כיתה יבש העצם </td><td> ט&quot;ו סמית </td><td> UN1013 </td></tr><tr><td> מרית </td><td> מרית מיקרו (14 ס&quot;מ) </td><td> פישר סיינטיפיק </td><td> 21-401-15 </td></tr><tr><td> הבדיקה פיירקס ספוט צלחות </td><td> 9-85 גם לנתח את הצלחת מ&quot;מ X 100 מ&quot;מ </td><td> תומאס מדעי </td><td> 7812G17 </td></tr><tr><td> פסטר טפטפות </td><td> סודה לימון </td><td> J &amp; H Berge </td><td> 71-5200-05 </td></tr><tr><td> מצבטים </td><td> סגנון # 5 </td><td> סיגמא </td><td> T-4662 </td></tr><tr><td> אתנול </td><td> 190 הוכחה USP </td><td> פישר סיינטיפיק </td><td> 04-355-221 </td></tr><tr><td> פורמלדהיד </td><td> 37% w / w </td><td> פישר סיינטיפיק </td><td> F79-1 </td></tr><tr><td> משני הנוגדנים Cy3 AffiniPure חמור נגד ארנב IgG (H + L) </td><td> 1:50 עירור 546 ננומטר, 565 ננומטר פליטה </td><td> ג&#39;קסון חיסונית מעבדות מחקר, Inc </td><td> 711-165-152 </td></tr><tr><td> Antifade (N-גאלאט) </td><td> 4 מיקרוגרם / מ&quot;ל ​​גליצרול 50% 1X PBS </td><td> מגה פיקסל Biomedicals </td><td> 10274790 </td></tr><tr><td> גליצרול </td><td></td><td> פישר סיינטיפיק </td><td> G33-1 </td></tr><tr><td> Hoechst 33258 </td><td> 0.2 מיקרוגרם / מ&quot;ל ​​עירור 352 ננומטר, 461 ננומטר פליטה </td><td> בדיקות מולקולריות </td><td> H-1398 </td></tr><tr><td> Rhodamine phalloidin </td><td> 200 יחידות / מ&quot;ל ​​(6.6מיקרומטר) עירור 540 ננומטר, 565 ננומטר פליטה </td><td> בדיקות מולקולריות </td><td> R415 </td></tr><tr><td> Alexa פלואוריד 488 phalloidin </td><td> 300 יחידות / מ&quot;ל ​​עירור 495 ננומטר, 518 ננומטר פליטה </td><td> בדיקות מולקולריות </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> לשטוף את הבקבוק </td><td> Fisherbrand </td><td> פישר סיינטיפיק </td><td> 03-409-22A </td></tr><tr><td> Kimwipes </td><td> קימברלי קלארק </td><td> פישר סיינטיפיק </td><td> 06-666A </td></tr><tr><td> נייר מגבת </td><td> 1 רובדי C פי </td><td> נוצה </td><td> 901-7CFTB2400 </td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td> מיקרוסקופיה </td><td> &amp;nbsp; </td><td></td><td></td></tr><tr><td> לייקה סטראו </td><td> MZFLIII </td><td> האימפריה Imaging Systems, Inc </td><td> 10446208 </td></tr><tr><td> Zeiss סטראו </td><td> STEMI 1000 או 2000-C </td><td> Carl Zeiss </td><td> 000000-1006-126 </td></tr><tr><td> מקור אור - LED </td><td> שולחן מתכווננת הפנס </td><td> פישר סיינטיפיק </td><td> 12563501 </td></tr><tr><td> שלב </td><td> תיבת אור מסור עם צלחת </td><td> Carl Zeiss </td><td> 455137000 </td></tr><tr><td> Zeiss סריקת לייזר מיקרוסקופ confocal </td><td> LSM 510 </td><td> Carl Zeiss </td><td></td></tr><tr><td> Zeiss תרכובת מיקרוסקופ </td><td> Axioplan 2 זקוף </td><td> Carl Zeiss </td><td></td></tr></tbody></table><table border="1" style=";text-align:right;direction:rtl"><tbody><tr><td> צרעה זנים </td><td> טוס מסנניםשל </td></tr><tr><td> Leptopilina victoriae 16 </td><td> י.ו. </td></tr><tr><td> Leptopilina boulardi 17 1 </td><td> כטב&quot;מ-GFP-הגבי 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> י.ו. הנחש-Gal4 כטב&quot;מ GFP-Dorsal/Basc 4 </td></tr><tr><td></td><td> י.ו.; 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

טפיל (טפילים) צרעות מהווים שיעור גדול של אויבים טבעיים של חרקים רבים, כולל<em&gt; תסיסנית melanogaster</em&gt;. נציג את הטכניקות הללו כדי להפיץ טפילים<em&gt; תסיסנית</em&gt; Spp. ולהדגים כיצד לנתח את השפעתם על המערכת החיסונית של רקמות<em&gt; תסיסנית</em&gt; זחלים.

מבוא של צרעות טפיליות<em&gt; תסיסנית</em&gt; ואת התגובה החיסונית Antiparasite

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

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Investigating the Effects of Probiotics on Pneumococcal Colonization Using an In Vitro Adherence Assay

חוקר את ההשפעות של פרוביוטיקה על דלקת ריאות התיישבות שימוש<em&gt; במבחנה</em&gt; Assay דבקות

Adherence of Streptococcus pneumoniae (the pneumococcus) to the epithelial lining of the nasopharynx can result in colonization and is considered a ...

Using RNA-interference to Investigate the Innate Immune Response in Mouse Macrophages

באמצעות RNA-התערבות לחקר התגובה החיסונית המולדת בהמקרופאגים עכבר

Macrophages are key phagocytic innate immune cells.  When macrophages encounter a pathogen, they produce antimicrobial proteins and compounds to kill the ...

The Utilization of Oropharyngeal Intratracheal PAMP Administration and Bronchoalveolar Lavage to Evaluate the Host Immune Response in Mice

הניצול של לוע התחתון Intratracheal PAMP מנהל ושטיפה ברונכואלוואולרית כדי להעריך את תגובת מארח חיסון של עכברים

The host immune response to pathogens is a complex biological process. The majority of in vivo studies classically employed to characterize host-pathogen ...

In Vitro Analysis of Myd88-mediated Cellular Immune Response to West Nile Virus Mutant Strain Infection

In Vitro Analysis of Myd88-mediated Cellular Immune Response to West Nile Virus Mutant Strain Infection

בניתוח חוץ גופייה של תיווך Myd88 הסלולרי תגובה החיסונית לוירוס הנילוס המערבי זיהום הזנים Mutant

An attenuated West Nile virus (WNV), a nonstructural (NS) 4B-P38G mutant, induced higher innate cytokine and T cell responses than the wild-type WNV in ...

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

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

הדמיה לא פולשנית של התגובה החיסונית המולדת בדגם דג הזברה הזחל של<em&gt; Streptococcus iniae</em&gt; זיהום

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

ייחסתי<em&gt; Gambiae אנופלס</em&gt; יתושים עם חרוזים כדי לעודד ו למדוד את התגובה החיסונית melanization

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