Progress 10/01/19 to 09/30/20
Outputs
Target Audience:Audiences were scientists and students in the Department of Entomology at TAMU through presentations in the Graduate Student Forum August 2020. Although Pietrantonio co-organized a symposium for the Entomological Congress of Entomology in Helsinki, Finland for July 2020, the congress was suspended until 2021 due to covid, so the research could not be presented as scheduled for her Oral presentation and the Abstract was not published. Pietrantonio continued collaborations through a trip in March to the CFTRL-USDA-ARS in Edinburg, Texas and conferenced with colleagues from the Kerrville, USDA-ARS. Target audiences were the Mosquito Vector Control community of CDC through the reports provided through the Western Gulf Center of Excellence in Vector Borne Diseases. Invitations to present were diminished due to covid measures. Changes/Problems:Covid affected all research activities since mid-March 2020, as follows: Cancelation of the International Congress of Entomology. Change in date for the CDC Annual meeting that was supposed to be scheduled in the Spring but was held virtually in October, outside of the period of this report. Lack of other opportunities to present our research in person. Cancellation of travel to the Edinburg USDA-ARS research facility and inhability to obtain live ticks for bioassays. A post-doc from Korea that had joined the lab in October 2019, Wook Cha, departed prematurely to Korea in June 2020 because of the COVID measures of isolation and stay at home regulations. She thought that under these circumstances in the USA, Korea had a lower transmission rate and left. This was a very big problem because Texas AgriLife Research prohibited ANY hiring with State funds so only we could hire personnel under extramural grants. Two of my grants were from the Texas Special Item from the legislature so I could not hire personnel. What opportunities for training and professional development has the project provided?One Hispanic undergraduate male (Carlos Ortega) continued to take research units as Senior, and was trained in molecular biology techniques: DNA extraction of H. zea, primer design, gene structural analyses with SnapGene online, primer optimization and agarose electrophoresis. He also wrote several reports which improved his technical writing skills. Upon graduation summer 2020 he obtained a job in a local Cancer Clinic. The three PhD students are fully funded including tuition and fees. This project provided excellent financial and technical opportunities. One PhD Graduate student C. Xiong learned High-Throughput screening obtaining excellent training. Hands on experience with robots for HTS is apar only to that available in facilities of few universities, mainly medical schools or big pharma. Theory training on validation of HTS was complemented through own experiential learning. New capital equipment purchased ensures a modern learning environment (new speedvac, milliq system, cell culture incubator, centrifuge for plates, new qPCR system, etc.). Caixing Xiong was nominated by Pietrantonio and obtained a MUVE section Travel Grant award the Entomological Society of America Annual Meeting on November 2019, and an Award ($100) for her research presentation from the Chinese association at the same meeting. She will graduate Dec. 2020. The other two students Jonathan Hernandez and Han-Jung Lee are fully funded through a Merit fellowship (JH) and by the CDC-WGCVBD (H-J. Lee). Both these students traveled several times to the Houston mosquito and vector control division in Harris County were they participated observing the field-control research trials from which they obtained mosquitoes for analyses of kdr mutations. H-J. Lee prepared a manuscript for publication submitted to PLoS Neglected Tropical Diseases. Both students learned molecular biology techniques, melting curve analyses, diagnostic PCR for single nucleotide polymorphisms and we develop a novel PCR of very simple application to detect pyrethroid resistance in Culex quinquefasciatus mosquitoes. Both students benefited with professional interactions with our collaborator and professor of Statistics Dr. M. Longnecker at TAMU. Students are always first authors if they performed the research experiments in the laboratory or in the field and are able to produce a first draft of the manuscript specifically on Materials and Methods and Results. The PD works side by side with students to help with their writing and recruits expert collaborators to participate in the diverse projects. Students interact and network with a multidisciplinary team of researchers and expand their network. The PD nominates students for Awards to develop their vita. Jonathan Hernandez obtained a 2nd place Award in the Graduate Student Forum competition, Dept. of Entomology, Aug. 2020, on this project on molecular characterization of kdr mutations in Ae. aegypti from Harris Co. How have the results been disseminated to communities of interest?The paper on the first prostaglandin receptor had been planned for dissemination at the Int. Congress of Entomology in Helsinki but this was postponed to 2021. Caixing Xiong presented her research at the ESA Nov. 2019 (this will be reported under the NIFA -AFRI federal grant that supported her financially). Dissemination was affected by COVID during 2020 as all trips to conferences were cancelled. Students presented in the 2020 Department of Entomology Graduate Student Forum that was also held virtually in August. Other dissemination is found under Other Products in the form of Reports. What do you plan to do during the next reporting period to accomplish the goals?Goal 1: we began collaboration with Dr. Aramayo (TAMU, Biology Dept.) and analyzed transcriptomes of ovaries from mated and virgin fire ant queens. We will continue analyzing and annotating these sequences. Goal 2. We have new experiments planed to determine the antifeedant effect of the kinin antagonists to be tested in mosquitoes. Goal 3. Work on resistance monitoring and diagnostics in mosquitoes is proceeding successfully and we will soon submita another publication on Aedes aegypti sodium channel of which the 2nd year PhD graduate student J.Hernandez will be the first author. For Cx. quinquefasciatus in Harris Conty (TX) we will develop methodology to correlate the amount of permethrin collected in the treatment zone with the field mortality of mosquitoes. This work is under preliminary methods development.
Impacts
What was accomplished under these goals?
Under Goal 2 with respect to: A calcium-fluorescence assay will be calibrated for high-throughput screens of small molecule libraries in 384-well plates. Chemical hits will be validated by dose-response in recombinant-receptor expressing cells. Subsequently candidate agonists will be tested in empty mammalian cells (only transfected with vector without receptor). Candidate antagonists and receptor-specific agonists will be tested in bioassays in vivo and in vitro. Most of these goals were accomplished. During this period we developed a dual addition fluorescence assay for discovery of agonists and antagonists during the same experiment and with the same cells. This was developed in my laboratory by my PhD student Caixing Xiong, and myself (Pietrantonio). In collaboration with Dwight Baker from the Dept. of Biochemistry and Biophysics we discovered small molecules antagonists of kinin receptors and specifically for this project we report the discovery of three small molecules that can significantly decrease the mosquito hindgut contraction elicited by a kinin agonist. This discovery has been submitted for publication. It is consider of high impact not only because we validated the function in blocking the receptor in vitro in recombinant cells but also demonstrating activity of these small molecules in a hindgut contraction inhibition assay indicating that the small molecules can have a physiological effect at the tissue level. Furthermore, we discover the pharmacophore group that appears crutial for antagonistic activity by using in silico screens of similar molecules to the most potent antagonist discovered as hit and validated in cells. This was accomplished by analyzing the activity of seven analogs with similar structures (SAR studies). A publication has been submitted. Under Goal 3. We published the discovery of the first insect prostaglandin receptor from the model lepidopteran Manduca sexta. This research impacts Insect Science overall, and specifically insect immunity in caterpillar pests. This has important impact on tools for biocontrol such as parasitoids, fungal entomopathogens and bacterial pathogens. It could represent a good target to diminish the defense response of insects to these pest control tools.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2020
Citation:
Kwon, Hyeogsun, Yunlong Yang, Sunil Kumar, Dae-Weon Lee, Prati Bajracharya, Travis L. Calkins, Yonggyun Kim, and Patricia V. Pietrantonio. Characterization of the first insect prostaglandin (PGE2) receptor: MansePGE2R is expressed in oenocytoids and lipoteichoic acid (LTA) increases transcript expression. Insect Biochemistry and Molecular Biology 117 (2020): 103290.
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Progress 11/01/18 to 09/30/19
Outputs
Target Audience:The PD has presented research in International, National, Regional and State and local TAMU venues in addition to teaching two graduate courses in Fall 2018 and Spring 2019, respectively. The PD received an International Travel invitation from the Animal and Plant Quarantine Agency (Korea) in April 2019, and the PD made two formal presentations in Korea, one at the APQA where the audience were quarantine professionals, professors and administrators, insect taxonomists, pest control researchers, insect physiology graduate students and technicians. The second presentation was at the Korean Society of Applied Entomology, Spring semester meeting in 2019 on the physiology of fire ants and the audience was mainly graduate students, postdocs and faculty from Korea. The PD also presented an invited oral presentation 8th International Symposium in Molecular Insect Science. Sitges, Spain. 7-10 July, 2019 where the audience was of international professors and researchers in molecular science of arthropods. The PD and her postdoc presented research at the Annual meeting of the Entomological Society of America in Vancouver, Canada, the audience being an international audience of professors and students, members of federal agencies such as NIFA and EPA, NIH, etc. This presentation was on fire ant queen brain transcriptomics Nov. 2018. The PD presented research at theConference of Research Workers in Animal Diseases (CRWAD) in Chicago 2-5 December, 2018, where the audience was of professionals, animal husbandry and veterinary pharma industry and students all working on animal diseases. The PD presented orally her research and participated in CDC meeting in Weslaco, TX, as part of the PD work on identification of mutations in the sodium channel of mosquito Culex quinquefasciatus. The audience were operational mosquito and vector control professionals, faculty from TAMU, UTRGV, UT El Paso, UTMB Galveston, Mosquito Control Units of Texas and administrators from Texas AgriLife Research, graduate students, post-docs, researchers and CDC external advisory panel and evaluators. Indeed the whole community of vector control in Texas was reunited for this Western Gulf Center of Excellence in Vector Borne Diseases annual meeting on June 20-21, 2019 at the University of Rio Grande Valley at Edinburg, TX.The PD is an advocate for women in Stem and was invited Panelist for the International Women's Day Conference at TAMU. The topic of the presentation wasWomen's leadership, broader impacts, diversity trends, women's rights, perspective from an international scholar in STEM. Invited by Dean Pamela Mathews. March 6, 2019. MSC, TAMU. The audience was mainly female, graduate students, undergraduate student, student organizations, student advocates, professors of various Departments and Colleges. Changes/Problems:As the postdocs have been successful in moving on and finding permantent jobs the challenge is to replace them with highly competent people. With the near full-employment market for qualified PhD personnel, this is a hard task, as our salaries cannot compete with those provided by private industry. There have been delays that originated with the government closure in February which caused our team to lose one animal experiment under the NIFA-AFRI grant, and determined that personnel had to switch emphasis on what they were planning on doing, disturbed planned expenditures, etc. What opportunities for training and professional development has the project provided?The project provided opportunities for 1 male postdoc, 1 female postdoc, 2 female PhD graduate students and 1 Hispanic undergraduate from the College of Sciences (Biology Dept.). The project also allowed a professor from Korea to spend 1 year of sabbatical in the laboratory and participating in the Manduca sexta prostaglandin receptor project. One female graduate student, Caixing Xiong, continued to acquire skills in chemical screens, analyzing data of highthroughput screens, validating assays through dose-response curves. The same student participated in the ESA Annual meeting in Vancouver, CA, and presented her research orally in the student competition. The Hispanic undergraduate is from an underserved population and this project provided him with the opportunity to complete the required research units from his undergraduate program in Biology. The female postdoc obtained a job and has moved on. Similarly Travis Calkins obtained a permanent job with the company Charles River in Ohio that specializes in toxicological animal models for facilitating and supporting pesticide registration. Another female PhD student, Han-Jung Lee had the opportunity to interact with the Harris County Vector Control Division and to participate in the Weslaco meeting of the Center of Excellence in Vector Borne Diseases. The PD is also the PD for Aim 1 of the Center of Excellence project (Director, Scott Weaver, UTMB). How have the results been disseminated to communities of interest?The PD has targeted many diverse communities during this period: The chemical industry and agrochemical researchers through her presentation at the IUPAC Congress in Ghent, Belgium; the animal workers community through the CRWAD conference in Chicago nationally, and internationally though her presentation in Sitges, Spain in July 2019. The mosquito and tick control community through the CDC-funded project as the PD belongs to the Western Gulf Center of Excellence in Vector Borne Diseases. Students at TAMU benefit constantly from the PD presenting her research findings in graduate courses, through laboratory group meetings, through her participation in Dept. Seminars and through her hosting of invited visiting scientists. What do you plan to do during the next reporting period to accomplish the goals?Goal 1. The main task is to replace Dr. Travis Calkins who had been working with fire ants. The PD has posted a job description to fill this position. Regardless, the PD collaborates with Dr Tamboringeguy and together have written a draft proposal to be submitted to NSF-IOS. the fire ants continued to be maintained in Pietrantonio lab. One challenge for the fire ant project this year was that there were not visible mating flights of the intensity seen in previous years. We successfully collected colonies in the field but failed to collect sufficient numbers of newly mated queens, as this cannot be mated in the laboratory, they must be found in parking lots after days of heavy rain and sunny, not windy conditions. The lack of newly mated queens have delayed some planned experiments. Goal 2. These experiments are under way and we do not anticipate failures at this point. A student is working on this project and the post doc helps part time. We will continue work as planned. Goal 3. Two other PhD students and one undergraduate are working on this goal and work is proceeding satisfactorily. We will continue work as planned.
Impacts
What was accomplished under these goals?
Goal 1. We published 3 journal articles. We accomplished the analysis of the fire ant queen brain and worker brain with respect to differentially expressed genes between mated and virgin queens, and between mated queen brain, virgin alate brain and workers. We discovered that a putative antimicrobial peptide, abaecin, is upregulated in queens after mating. We annotated the fire ant G protein-coupled receptors and summarized this in an invited journal article. We produced a list of the GPCRs found in the fire ant genome and reannotated some of them. We also performed phylogenetic analyses of these receptors and of the G proteins. All of these findings advance neurobiology and provided a list of curated GPCRs to complement our studies. Goal 2. We successfully developed a 384-well plate assay for highthroughput screens. We developed methodology for analyzing the potency and efficacy of peptidomimetics that were satisfactorily tested in recombinant cell expression assays (in CHO-K1 cells). A regression line and correlation analysis was obtained for efficacy vs potency for analogs that were active in both mosquito and tick receptors. We are conducting pilot studies of tick larvae by immersion and we are testing several commercially available solvents utilized in agrochemicals as we evaluate them for tick toxicity. We published our approaches and ideas in the Current Opinion on Insect Science invited paper and we published several papers on this topic on ticks, many publications will be reported under the NIFA-AFRI grant that is current, as to avoid overlap. Pietrantonio received several international invitations for oral presentations and invited publications in peer reviewed journals on research focused on this goal. Goal 3. Complementing our studies with H. zea, bollworm, we continued research on lepidopteran immunity focusing on the prostaglandin receptor from the model lepidopteran Manduca sexta. We submitted a manuscript that was under revision. This is the first prostaglanding receptor characterized in any insect or arthropod. We continued our studies on the H. zea larvae on mechanisms of resistance to Bt toxins present in transgenic crops (cotton and corn). We submitted a NIFA-AFRI grant during the summer of 2019 in collaboration with other TAMU researchers on the sugar cane aphid.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2018
Citation:
Calkins, T., M.-E. Chen, A. Arora, C. Hawkings, C. Tamborindeguy, P. V. Pietrantonio. 2018. Brain gene expression analyses in virgin and mated queens of fire ants reveal mating-independent and socially-regulated changes. Ecology and Evolution 8: 4312-4327.
- Type:
Journal Articles
Status:
Published
Year Published:
2018
Citation:
Pietrantonio, P.V., C. Xiong, R. J. Nachman and Y. Shen. 2018. G protein-coupled receptors in arthropod vectors: Omics and pharmacological approaches to elucidate ligand-receptor interactions and novel organismal functions. Current Opinion in Insect Science 29:12-20.
- Type:
Journal Articles
Status:
Published
Year Published:
2019
Citation:
Calkins, T., C. Tamborindeguy, and P. V. Pietrantonio. 2019. GPCR annotation, G proteins and transcriptomics of Solenopsis invicta queen and worker brain: an improved view of signaling in an invasive superorganism. Gen. Comp. Endocrinol. 278:89-103. Invited Special Issue.
- Type:
Journal Articles
Status:
Published
Year Published:
2019
Citation:
Hawkings C, Calkins TL, Pietrantonio P.V., Tamborindeguy C. 2019. Caste-based differential transcriptional expression of hexamerins in response to a juvenile hormone analog in the red imported fire ant (Solenopsis invicta). PLoS One. 2019 May 20;14(5):e0216800. DOI: 10.1371/journal.pone.0216800
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Pietrantonio, P.V. 2019. A superorganism on the move: will social endocrinology help stop the global spread of the fire ant? Invited to the Special Symposium organized by the Animal and Plant Quarantine Agency (Korea) Prevention of Global Spreading of a Red Imported Fire Ant . 2019 Spring International Conference of the Korean Society of Applied Entomology (KSAE). Cheongju, Korea. 24-26 April 2019. Page 13.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Pietrantonio, P.V., C Xiong, R.J. Nachman. 2019. Evaluation of Aib and PEG-polymer insect kinin analogs on mosquito and tick GPCRs identifies potent new pest management tools with potentially enhanced biostability and bioavailability. 14th IUPAC International Congress of Crop Protection Chemistry. Ghent, Belgium. May 19-23, 2019.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
Calkins, T.L., Tamborindeguy, C., and P.V. Pietrantonio. 2018. Mating and socially regulated changes in fire ant queen brain transcriptomes. Entomological Society of America Annual meeting, Vancouver, BC, Canada. November 13, 2018.
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