Name: an introduction to parasitic wasps of drosophila and the antiparasite immune response
Uploaded: 2012-05-07T12:00:00
Description: Watch this Scientific Journal Video about An Introduction to Parasitic Wasps of Drosophila and the Antiparasite Immune Response at JoVE.com

우리는 Trichopria drosophilae, 유전자 변형 뜬 종자를위한 교수 토니의 IP, 그리고 안티 Spätzle 항체에 대해 교수 칼 하시 모토위한 교수 토드 Schlenke에 감사를드립니다. 이 프레 젠 테이션에 대한 그들의 공헌에 대한 연구소의 현재와 과거 회원을 감사드립니다. NIH (S06 GM08168, 대두 41399-009, 그리고 G12-RR03060), USDA (NRI / USDA CSREES 2006-03817하고 2009-35302-05277) 및 PSC-CUNY에서이 작품은 다음과 같은 보조금에 의해 지원되었다.

실험에 대한 전체 프로토콜은 네 단계 (그림 9)으로 나누어져 있습니다. 파리 유충에 (1) Culturing의 말벌 (2) 감염을 설정 및 해부를 위해 동물을 준비 (3) 분리 및 호스트 / 기생충 구조를 고치고, (4) 면역 조직 분석. 1. Drosophila의 애벌레에 Culturing의 말벌 말벌 문화의 유지 보수 신중한 계획이 필요합니다. 성장하는 파리에 대한 상대, 그것?...

<ol>
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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=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>

Drosophila의 기생 말벌에 대한 관심은 전체 genomes를 해독하기위한 분자 기술은 효율적이고 비용 효과가로 서징있다. 그러나 그들의 매우 잘 공부 호스트에 상대적으로, 말벌 생물학의 많은 매혹적인 측면이 모호한 상태로 유지됩니다. 이러한 범위, 면역 억제, superparasitism, 그리고 행동을 호스팅과 관련된 문제가 포함됩니다. 이 프레 젠 테이션의 초점은 파리의 면역 조직에서 감염의 효과를 ?...

<table border="1"><tbody><tr><td> 자료 </td><td> 유형 </td><td> 회사 </td><td> 카탈로그 번호 </td></tr><tr><td> 곤충의 문화 유지를위한 자료 </td><td></td><td></td><td></td></tr><tr><td> 누룩 </td><td> 활성 건조 </td><td> 피셔 과학 </td><td> S802453 </td></tr><tr><td> 음식 들아 </td><td> 옥수수 식사, 설탕 </td><td></td><td> 표준 제조법 </td></tr><tr><td> 꿀 </td><td> 클로버 </td><td> 네덜란드 골드 </td><td></td></tr><tr><td> 튜브 </td><td> 폴리 프로필렌 셸 튜브 (좁은) </td><td> 피셔 과학 </td><td> AS514 </td></tr><tr><td> 유리병에 폐쇄 </td><td> 코튼 플러그 </td><td> 피셔 과학 </td><td> AS212 </td></tr><tr><td> 유리병에 폐쇄 </td><td> 버즈 플러그 </td><tD&gt; Genesee 과학 </td><td> AS273 </td></tr><tr><td> 냉장 인큐베이터 </td><td> 정밀 815 </td><td> 써모 과학 </td><td> 3721 </td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td> 샘플 준비를위한 자료 </td><td></td><td></td><td></td></tr><tr><td> CO 2 탱크 </td><td> 뼈 건조 학년 </td><td> TW 스미스 </td><td> UN1013 </td></tr><tr><td> 주걱 </td><td> 마이크로 주걱 (14 ㎝) </td><td> 피셔 과학 </td><td> 21-401-15 </td></tr><tr><td> Pyrex 명소 테스트 플레이트 </td><td> 9 - 잘 해부 판 85mm X 100mm </td><td> 토마스 과학 </td><td> 7812G17 </td></tr><tr><td> 파스퇴르 Pipettes </td><td> 소다 석회 </td><td> J &amp; H Berge </td><td> 71-5200-05 </td></tr><tr><td> 집게 </td><td> 스타일 # 5 </td><td> 시그마 </td><td> T-4662 </td></tr><tr><td> 에탄올 </td><td> 190 증거의 USP </td><td> 피셔 과학 </td><td> 04-355-221 </td></tr><tr><td> 포름 알데히드 </td><td> 37% W / w </td><td> 피셔 과학 </td><td> F79-1 </td></tr><tr><td> 이차 항체 Cy3 AffiniPure 당나귀 안티 토끼 IgG (H + L) </td><td> 1시 50분 여진 546 nm의; 배출 565 nm의 </td><td> 잭슨 Immuno 리서치 연구소 주식 회사 </td><td> 711-165-152 </td></tr><tr><td> Antifade (N-프로필 gallate) </td><td> 1X PBS에 50 % 글리세롤 4 μg / ML </td><td> MP의 Biomedicals </td><td> 10274790 </td></tr><tr><td> 글리세린 </td><td></td><td> 피셔 과학 </td><td> G33-1 </td></tr><tr><td> Hoechst 33258 </td><td> 0.2 μg / ML 여기 352 nm의; 배출 461 nm의 </td><td> 분자 프로브 </td><td> H-1398 </td></tr><tr><td> Rhodamine phalloidin </td><td> 200 개가 / ML (6.6μm의) 여진 540 nm의; 배출 565 nm의 </td><td> 분자 프로브 </td><td> R415 </td></tr><tr><td> 알렉사 형석 488 phalloidin </td><td> 300 단위 / ML 여기 495 nm의; 배출 518 nm의 </td><td> 분자 프로브 </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> 병을 씻으십시오 </td><td> Fisherbrand </td><td> 피셔 과학 </td><td> 03-409-22A </td></tr><tr><td> Kimwipes </td><td> 킴벌리 클라크 </td><td> 피셔 과학 </td><td> 06-666A </td></tr><tr><td> 종이 타월 </td><td> 하나 플라이 C-공간 </td><td> 깃 </td><td> 901-7CFTB2400 </td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td> 현미경 사용법 </td><td> 및nbsp; </td><td></td><td></td></tr><tr><td> Leica의 stereomicroscope </td><td> MZFLIII </td><td> 제국 이미징 시스템즈 </td><td> 10446208 </td></tr><tr><td> 자이스 혈구 Stereomicroscope </td><td> Stemi 1000 또는 2000-C </td><td> 칼 자이스 혈구 </td><td> 000000-1006-126 </td></tr><tr><td> 광원 - LED </td><td> 거위 목처럼 생긴 것의 조명기 </td><td> 피셔 과학 </td><td> 12563501 </td></tr><tr><td> 무대 </td><td> 플레이트로 전송 라이트 박스 </td><td> 칼 자이스 혈구 </td><td> 455,137,000 </td></tr><tr><td> 자이스 혈구 레이저 공촛점 현미경 스캐닝 </td><td> LSM 510 </td><td> 칼 자이스 혈구 </td><td></td></tr><tr><td> 자이스 혈구 복합 현미경 </td><td> 직립 Axioplan이 </td><td> 칼 자이스 혈구 </td><td></td></tr></tbody></table><table border="1"><tbody><tr><td> 말벌의 변종 </td><td> 스트레인 날기의 </td></tr><tr><td> Leptopilina victoriae 16 </td><td> yw </td></tr><tr><td> Leptopilina boulardi 17 1 </td><td> UAS-GFP-지느러미 17 </td></tr><tr><td> Leptopilina heterotoma이 </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 뱀의-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).

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

Parasitoid (기생) 말벌을 포함한 여러 곤충들의 천적의 주요 클래스를 구성<em&gt; Drosophila melanogaster</em&gt;. 우리는 이러한 기생충을 전파하는 기법을 소개합니다<em&gt; Drosophila</em&gt; SPP. 과 면역 조직에 미치는 영향을 분석하는 방법을 보여줍니다<em&gt; Drosophila</em&gt; 애벌레.

의 기생충 말벌에 대한 소개<em&gt; Drosophila</em&gt; 그리고 Antiparasite 면역 반응

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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Quantification of the Respiratory Burst Response as an Indicator of Innate Immune Health in Zebrafish

제브라 피쉬의 선천성 면역 건강의 지표로 호흡 버스트 응답의 정량화

The phagocyte respiratory burst is part of the innate immune response to  pathogen infection and involves the production of reactive oxygen species (ROS). ...

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

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

를 사용하여 폐렴 구균 식민지에서 생균제의 효과를 조사<em&gt; 체외</em&gt; 준수의 분석

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

마우스 대 식세포의 타고난 면역 반응을 조사하기 위해 RNA 간섭을 사용하여

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

마우스의 호스트 면역 반응을 평가하기 위해 구인두 기관 내 PAMP 관리 및 폐포의 활용

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

웨스트 나일 바이러스 돌연변이 균주의 감염에에서 MyD88 매개 세포 면역 반응의 생체 외 분석

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

의 Zebrafish의 애벌레 모델의 선천성 면역 반응의 비 침습적 영상<em&gt; 연쇄상 구균 iniae</em&gt; 감염

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

접종<em&gt; 아노 펠 레스 gambiae</emMelanization 면역 반응을 유도하고 측정하는 비즈&gt; 모기

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