Progress 02/01/09 to 07/31/12
Outputs
OUTPUTS: 1. Taught and mentored postdoc, graduate students, undergraduate students, and technicians in classes and in the laboratory. Three PhD students and 2 Masters students completed their research thesis and graduated during this period. Two of the 5 graduate students are minority students. One of them is pursuing a postdoctoral fellowship at NYU and is considering a tenure-track job offer at an academic institution. The other one is a student at Baylor Medical College. 2. Presented findings on "Acute inflammation and anti-inflammation in Drosophila larval hosts: Lessons from parasitc wasps" in 12th RCMI International Symposium on Health Disparities, December 6-9, 2010 in Nashville, Tennessee. In 2011 and 2012, attended the Annual Drosophila conference in San Diego and Chicago, respectively. 3. Released database on Immunity genes expression in Drosophila. The database paper is published and it has more than 55 registered users world-wide. The link for the database is http://134.74.112.41/PADMA/. The publication is listed under the publication list. 4. In unpublished results, we have shown that viral ankyrin molecules from polyDNA viruses infecting plant pest Heliothes virescens acts to block the Toll pathway in Drosophila embryos and larvae. 5. In collaboration with Dr. Vladimir Gokhman (Moscow State University), a renowned karyologist of parasitic wasps, we have studied karyotypes of four parasitic wasp species that attack Drosophila. FISH experiments are ongoing to map genes to chromosomes of L. heterotoma. We have also determined their genome sizes (see publication below: Gokhman et al., 2011). We are pursuing sequencing of the L. heterotoma genome via the i5K initiative (http://arthropodgenomes.org/wiki/i5K). 6. We have developed Drosophila and its natural parasitic wasps as a model to understand host immune responses and inflammation in insects. We are also interested in understanding how natural parasites evade or suppress the immune responses of the fly hosts. To this end, we are characterizing the proteins of immune-suppressive virus-like particles (developing computational models & transgenic strains) to study their effects on the fly immune system. These studies are revealing the nature of virulence mechanisms underlying high success of parasitoids. This work will provide new modalities for insect pest control. 7. We have published 9 papers including 1 book chapter during this period. We are preparing three more papers for publication this year. The PD was invited to join the Honorary Editorial Advisory Board of the new journal Invertebrate Immunity, being launched by Springer (www.springer.com) and Versita (www.versita.com) PARTICIPANTS: Personnel in the lab during the reporting period: Postdoc: Gwenaelle Gueguen. Technicians: Zoe Papadopol & Roma Rajwani. Graduate students: Marta Kalamarz, Indira Paddibhatla, Chiyedza Small, Johnny Ramroop, Mark Lee, Maria Otazo. Undergraduate students Jeffrey Uribe,Tamara Goncharuk, Bodunde Onemola, Jemila Caplan Kester,Shuhebur Rahman,Shalena Islam, Chitra Chand, Arielle Scardino, Lawrence Huang, Rebecca Moore. Collaborations with Shireen Saleque (CCNY), Shaneen Singh (Brooklyn College), Vladimir Gokhman (Moscow State University), Eric Lai (Memorial Sloan Kettering Cancer Center, Ravi Sachidanandam (Mount Sinai School of Medicine), Rong wang (Mount Sinai School of Medicine). The PI wrote several grants during this period and was funded an award from NSF for the duration of 9/1/11-8/31/14 Molecular and functional analysis of virulence factors from L. heterotoma, a generalist parasite of Drosophila ($499,270). The USDA grant also allowed the generation of other data on inflammatory and detoxification genes that are currently under investigation for submitting new grants. Two small grants are under review. The PI and students got opportunities to travel to conferences and present data. TARGET AUDIENCES: The JoVE paper (Small et al., 2012) will share techniques to culture parasitoid wasps on flies. This has become a popular model system as many labs have requested wasp strains from the PD's lab. The PD was invited to give the following seminars: Rutgers University, Department of Genetics, May 12, 2010 (host Dr. Mary Konsolaki) Inflammation and immune homeostasis in Drosophila Indian Institute of Science Education Research, Pune, Dept of Biology July 27, 2011 (host Dr. G. Ratnaparkhi) Sumoylation enzymes and cancer inflammation: Studies in the Drosophila model organism. Chinese Academy of Sciences, Beijing, Institute of Zoology August 11, 2011 (host Dr. Cheng-Zhu Wang) VLPs from Leptopilina and their immune-suppressive properties in Drosophila spp Center for Cellular and Molecular Biology, Hyderabad, January 10, 2012 Department of (host Dr. Rakesh Mishra) The origin and growth dynamics of inflammatory hematopoietic microtumors of Drosophila: A new model for cancer inflammation The Program Director invited the following speakers to CCNY for a seminar: Iva Greenwald, Columbia University, Fall2010 (LIN-12/Notch signaling in C. elegans). Sean Megason, Harvard University, Spring 2010 (Deciphering the execution of the genomic code), Fred Berkovitch, Harvard University, Spring 2010 (Research opportunities at systems biology at Harvard). Johan Paulsson, Harvard University, Spring 2010 (Randomness in biology) Daniel Kalderon, Columbia University, Fall 2011 (Investigating a model adult stem cell: The Drosophila follicle stem cell) Mary Beth Saffo, National Science Foundation, Spring 2012(A mutualistic symbiosis with parasitic origins: probing a marine menage a trois) Please also see Output and Impact PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
1. We have examined the effects of various doses of nitenpyram on Drosophila viability. 2. We have determined the sublethal dose of nitenpyram on Drosophila. 3. We have found that there is no effect of the sublethal dose of this drug on the development of parasitoid-infected Drosophila. 4. We have acquired natural microbial pathoges of Drosophila (virulent strains of Pseudomonas and Erwinia carotovara caratovara) to examine if the sublethal doses of nitenpyram synergize with these pathogens and compromise host health. 5. We have characterized the acute inflammatory response in fly larvae in response to parasitoid oviposition. A regulatory circuit with feed forward and feedback loops was identified in this study. This study, published in 2010 (Paddibhatla et al.) in PLoS Pathogens, has total article views > 2,800 and 700 downloads. Paddibhatla et al. 2010 paper was also picked up by the media. Some links are below: http://www.sciencedaily.com/releases/2011/01/110103134104.htm http://press-news.org/17583-parallels-between-cancers-infection-suppr ession-reported.html http://www.brightsurf.com/news/headlines/61770/Parallels_Between_Canc ers_Infection_Suppression_Reported.html 6. We have shown that loss of regulation of the circuit (item 5) leads to the development of hematopoeitic inflammatory microtumors. These tumors originate from the hematopoietic stem-like population of the larval lymph gland. Please see publication Kalamarz et al., 2012. 7. In a 2011 paper in Virus research, we showed that VLPs from the virulent L. boulardi-17 (Lb-17), that can neither lyse blood cells nor suppress host defense, undergo biogenesis similar to that of L. heterotoma. Like L. heterotoma VLPs, Lb-17 VLPs are stellate; but they have fewer spikes, each spike being significantly longer than the spikes in L. heterotoma VLPs. 8. We discovered the presence of Wolbachia in Leptopilina victoriae. We also found that it induces cytoplasmic incompatibility. Please see Gueguen et al., submitted. 9. The PD's lab was invited by Journal of Video Experiments to share techniques of growing wasps and studying immune tissues in Drosophila larvae. This paper is in press (Small et al., 2012).
Publications
- Lee, M.J., Mondal, A., Small, C., Paddibhatla, I., Kawaguchi, A., Govind S. (2011) A database for the analysis of immunity genes in Drosophila: PADMA database. Fly (Austin) 5 (2): 155-161. PMID: 21273816
- Gueguen G., Rajwani R., Paddibhatla I., Morales J., Govind S. (2011) VLPs of Leptopilina boulardi share biogenesis and overall stellate morphology with VLPs of the heterotoma clade. Virus Research 160(1-2):159-65.
- Gokhman, V.E., Johnston, J.S., Small, C., Rajwani, R., Hanrahan, S.J., Govind, S. (2011) genomic and karyotypic variation in Drosophila parasitoids (Hymenoptera, Cynipoidea, Figitidae). Comparative Cytogenetics 5(3) 211-221.
- Kalamarz, M., Paddibhatla, I., Nadar, C., Govind, S. (2012) Sumoylation is tumor-suppressive and confers proliferative quiescence to hematopoietic progenitors in Drosophila melanogaster larvae. Biology Open 1:161-172; doi: 10.1242/bio.2012043.
- Ferrarese, R., Morales, J., Fimiarz, D., Webb, B.A., and Govind, S. (2009) A supracellular system of actin-lined canals controls biogenesis and release of virulence factors in parasitoid venom glands. Journal of Experimental Biology 212: 2261-2268. PMID: 19561216. PMCID: PMC2702457
- Tsuyoshi, T., Sorrentino, R.P., Russell, M., Ferrarese, R., Govind, S., Schulz, R.A. (2009) Characterization of a lamellocyte transcriptional enhancer located within the misshapen gene of Drosophila melanogaster. PLoS One 4(7):e6429. PMID: 19641625. PMCID: PMC2713827.
- Paddibhatla I., Lee M.J., Kalamarz M.E., Ferrarese R., Govind S. (2010) Role for sumoylation in systemic inflammation and immune homeostasis in Drosophila larvae. PLoS Pathogens 6(12):e1001234. PMID: 21203476
- Small, C., Paddibhatla, I., Rajwani, R., Govind, S. (2012) An introduction to parasitic wasps of Drosophila and the antiparasite immune response. Journal of Visualized Experiments: Immunology and Infection. Manuscript # 3347. In press.
- Gueguen G., Bodunde O., Govind S. Association and Effects of Wolbachia on Leptopilina victoriae, a virulent parasitic wasp of Drosophila spp. (2012) Submitted to Applied and Environmental Microbiology. Manuscript # AEM01058-12
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Progress 02/01/10 to 01/31/11
Outputs
OUTPUTS: We have examined the effects of various doses of nitenpyram on Drosophila viability. We have determined the sublethal dose of nitenpyram on Drosophila. For the preliminary experiments combining insecticide and pathogens, we choose to use three insecticide doses, respectively killing 15% (DL15), 30% (DL30) and 50% (DL50) of the flies in the condition we used. Second instar larvae (3 to 4 day-old) of D. melanogaster were fed with a mixture of regular fly food and one of the three doses of insecticide and infected with the wasp Leptopilina victoriae for 48 hours. 24h after the wasps were removed, larvae were dissected and the infection, total encapsulation and efficient encapsulation rates were recorded. The insecticide seems to affect both total and efficient encapsulation rates when the dose applied is higher than DL15. There also seems to be a tendency toward a higher variation in the mean number of wasp egg per larvae with increasing doses of insecticide. The same protocol was used but for only DL15 and DL30 and the same parameters as for D. melanogaster were recorded. In this case the interpretation is less straightforward. The total encapsulation rate appears to drop with the lower dose used (DL15) but then comes back to the level of the controls when DL30 was applied. The same pattern is observed for the mean number of egg per larva. This number is similar in controls and DL30 but is increased for DL15. For insecticide and bacterial infection experiments, we used the same insecticide (nitenpyram) at a the DL30 dose and 2 different species of bacteria: Pseudomonas insecticola (Pe) and Erwinia caracatova caracatova 15 (Ecc). The insecticide and or the bacterial cultures were mixed with crushed banana and were given to 5 day-old D. melanogaster larvae for 3 hours, at 29C. The whole mixture was then put on regular fly food and the emergence of flies was recorded everyday for each condition. As expected Pe has a stronger effect on D. melanogaster survival than Ecc. The insecticide also gives the expected result as around 65% of the flies survived. When each bacterium was combined with the insecticide, the fly survival dropped dramatically. In the course of these experiments, we discovered that our strain of L. victoriae is infected with Wolbachia of the B group which is closely related to a Wolbachia strain of L. clavipes and Nasonia vitripennis. In L. victoriae, Wolbachia manipulates its host's reproduction by inducing "male development" type of cytoplasmic incompatibility. Measures of a number of life history traits reveal a slight effect of Wolbachia on L. victoriae size, developmental time, and fecundity. While L. victoriae cannot succeed on D. hydie, D. willistoni, D. virilis, and D. pseudobscura, Wolbachia does not affect its ability to attack and infect D. melanogaster, D. yakuba and D. simulans. The experiments were done by a postdoc, graduate students, and technicians. All members presented data in lab meeting where they received detailed feedback from the PD and specific mentoring in writing and presentation skills. PARTICIPANTS: Dr. Shubha Govind (PD) Oversee research, train students and postdocs at all levels; initiate and finalize writing, communicate and disseminate research results, oversee collaborations, administrative tasks with respect to grants and contracts. Dr. Gueguen, Gwenaelle (post-doctoral fellow) Perform research in collaboration with undergraduate and students. Write manuscript drafts and prepare figures for publication. Participate in reading literature and presenting data in lab meeting. Help with mentoring younger students. Ms. Zoe Papadopol (technician) Prepare culture media. Help maintain fly strains in the lab meticulously. Ms. Indira Paddibhatla (graduate student) Perform research in collaboration with undergraduate student/technician. Write manuscript drafts and prepare figures for publication. Participate in reading literature and presenting data in lab meeting. Ms. Chiyedza Small (recent PhD) Perform research in collaboration with undergraduate/MA student. Write manuscript drafts and figures for publication. Participate in reading literature and presenting data in lab meeting. Ms. Roma Rajwani (technician) Assist with all activities in the lab including growing wasps, ordering, keeping track of invoices, and performing experiments, when necessary. Collaborators and contacts Vladimir E. Gokhman Moscow State University Analysis of karyotypes of Drosophila wasps J. Spencer Johnston Texas A and M University Analysis of genome sizes of Drosophila wasps Matthew L. Buffingron Systematic Entomologist, ARS-USDA and Smithsonian Museum of Washington Shaneen Singh Brooklyn College of CUNY Modeling of proteins Todd Schlenke Emory University Exchange ideas, fly strains, and species TARGET AUDIENCES: City College is a minority and hispanic-serving institution. The PD is teaching undergraduate genetics to over 150 students during the academic year. She therefore has a strong impact on a significant component of the student population that will declare Biology as a major. Laboratory experience is brought into the classroom, as necessary. The Program Director's laboratory also has numerous undergraduate and graduate students from the under-represented minority communities. ALl students are mentored in the research methods, ethics and methods in a systematic and routine manner. In addition, the Program Director conducts teaching at CCNY. The class room has a diverse group of students. Students are introduced to the state-of-the art technologies and methods developed in the lab or via the World-Wide-Web. PROJECT MODIFICATIONS: In collaboration with Dr. Matt Buffington, The PD has proposed whole genome sequencing of Ganaspis and Leptopilina spp. This possiblity became possible because Dr. Gokhman has visited the PDs lab to study karyotypes of wasps. These experiments are leading to the development of methods for FISH analysis, that will be necessary for physical mapping of genes on chromosomes. The laboratory is also investigating the function of peroxidasin pxt in insect immunity. These new directions do not alter the original goals of the project.
Impacts
Events -Three graduate students attended 51st Annual Drosophila conference in Washington DC. They also attended workshops on specific topics held during the workshop. -Postdoctoral fellow attended the 12th RCMI International Symposium on health Disparities December 6, 2010. Dissemination developed and published a database for immunity genes in Drosophila. Data from our unpublished experiments. See reference on PADMA under publications.
Publications
- Gueguen, G., Rajwani, R., Paddibhatla, I., Morales, J., Govind, S. (2011) VLPs of Leptopilina boulardi Share Biogenesis and Overall Stellate Morphology with VLPs of the heterotoma Clade. Submitted to Virus research. Ms. Ref. No.: VIRUS-D-11-00199
- Kalamarz, M., Paddibhatla, I. and Govind S. (2011) Sumoylation is Tumor-Suppressive and Confers Proliferative Quiescence to Hematopoietic Progenitors in Drosophila melanogaster Larvae. Submitted to Development. MS ID#: DEVELOP/2010/062596
- Gokhman, V.E., Johnston, J.S., Small, C., Rajwani, R., Hanrahan, S.A., Govind, S. (2011) Genomic and Karyotypic Variation in Drosophila Parasitoids. Submitted to Comparative Cytogenetics.
- Abstracts 2010
- Paddibhatla, I., Lee, M.J., Ferrarese, R., Kalamarz, M.E., Govind, S. (2010) Ubc9-dependent modulation of NF-kB-dependent inflammation in Drosophila. Ann. Dros. Res. Conf. 411C. Washington DC.
- Gueguen, G., Paddibhatla, I., Rajwani, R., Govind, S. 2010. Inflammation and anti-inflammation in Drosophila larval hosts: Lessons from parasitic wasps. 12th RCMI International Symposium on health Disparities December 6, 2010. Abstract book. 07.01.37.
- Paddibhatla I., Lee M.J., Kalamarz M.E., Ferrarese R., Govind S. (2010) Role for Sumoylation in Systemic Inflammation and Immune Homeostasis in Drosophila Larvae. PLoS Pathogens 6(12):e1001234. PMID: 21203476
- Lee, M.J., Mondal, A., Small, C., Paddibhatla, I., Kawaguchi, A., Govind S. (2011) A Database for the Analysis of Immunity Genes in Drosophila: PADMA Database. Fly (Austin);5(2) 155-161.
- Small, C. Paddibhatla, I, Rajwani, R. and Govind, S (2011) An Introduction to Parasitic Wasps of Drosophila and the Antiparasite Immune Response. Journal of Video Experimentation. Accepted.
- Submitted 2010-2011
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