Progress 10/01/15 to 09/30/20
Outputs
Target Audience:The audience of our studies is constituted of biochemists, biologists, and entomologists that are interested in the metabolism of fat in vertebrates and in arthropods. The audience of our studies is distributed around the world in research institutions. In this reporting period (2016-2020) our studies were cited in approximately 1,600 articles (Information obtained from Google Scholar). Graduate and undergraduate students in our department also constitute an important audience for our studies. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Six undergraduate students worked for significant periods of time (more than a semester) on the project. Three graduate students worked on this project. One of them graduated with a Ph.D. in Biochemistry. The second graduate student completed and defended his Masters's thesis in 2018. He continues working on the project with the intention of earning a Ph.D. degree. The third student started working on the project in 2018 with the goal of earning a degree of Masters in Biochemistry. She continues working on the project. How have the results been disseminated to communities of interest?Some of the results obtained in this period were disseminated to the scientific community in the form of peer-reviewed publications. Other studies already completed will be reported in 2021 and beyond. Moreover, the students that participated in the project presented their results to different OSU audiences, some from our department and some to broader audiences, such as those attending theannual BMB-Graduate Student Association Symposium and the Niblack Research Scholars Symposium. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
Our contribution to the annotation of the genome of the Tobacco hornworm (Manduca sexta)was an important accomplishment. Our specific role was the identification and correction of the genomic sequences of the genes involved in lipid metabolism in this insect. Our work led to the identification of genes of lipid-binding proteins, acyltransferases, lipases, lipid droplet proteins, and other relevant proteins involved in the synthesis or in the degradation of fat in insects. The genome of Manduca sexta is a key tool needed to advance the understanding of insect physiology. Regarding the regulation of TG accumulation/degradation, we had previously shown in in vitro and in vivo studies that PLIN1A was involved in the activation of the hydrolysis of TG in fat body. In the current period, we made two additional advances related to the regulation of TG storage First, we learned and reported that PLIN2 is expressed and needed when there is an active synthesis of TG disregard of whether the fat body is accumulating or using and losing TG. Previous studies in Drosophila led to the notion that PLIN2 would help in the accumulation of TG reserves by preventing access of the lipases to the surface of lipid droplets and, thus, also preventing the hydrolysis and mobilization of TG. However, we have shown that the highest expression level of PLIN2 in fat body is observed when the tissue is in the highest lipolytic state. Our results suggested that PLIN2 does not prevent the hydrolysis of TG. PLIN2 would be needed during the assembly of small, nascent, lipid droplets. Moreover, a new isoform (isoform-B) of the insect perilipin 1, PLIN1, was identified in Manduca sexata and reported. PLIN1B appears to be specific to Lepidoptera. The nutritional status affects the expression of PLIN1B, However, expression of PLIN1B is limited to the last larval instar (5th) and reaches a maximum at the time of maximum accumulation of TG. Unlike PLIN1 A, whose expression correlates with a lipolytic state, PLIN1 B could be involved in maximizing TG accumulation.
Publications
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Progress 10/01/18 to 09/30/19
Outputs
Target Audience:The audience of our studies is constituted by biochemists, biologists, and entomologists that are interested in the metabolism of fat invertebrates and in arthropods. The audience of our studies is distributed around the world in research institutions. Our studies were cited in approximately 300articles reported during this 2018-2019period (Information obtained from Google Scholar).Graduate and undergraduate students in our department also constitute an important audience for our studies. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Two graduate students worked and are still working in the lab. This August, Karen Belloso Rojas started working in the project with the goal of earning a degree ofMasters in Biochemistry. The second student, Claudio Galvez Sagastume, whocompleted his M.S. in 2018continues working in the project with the intention of earning a Ph.D. degree. How have the results been disseminated to communities of interest?We reported some of the studies on the PLIN1 protein in the international journal Insect Biochemistry and Molecular Biology (This is considered the most important journal for insect biochemistry). Moreover, the students presented the results of their studies to different OSU audiences, some from our department and some diverse audiences, such as those attendingthe annualBMBGSA Symposium and the Niblack Research Scholars Symposium. What do you plan to do during the next reporting period to accomplish the goals?We will continue studying the roles of some genes involved in the metabolism of fat in insects. We will focus on the roles of the lipid droplet specific proteins (PLIN1 and PLIN2) in the accumulation of lipids in fat body andovaries. We will submit for publicationin specialized journals thestudies on the roles of apoLp and LTP in lipid metabolism.
Impacts
What was accomplished under these goals?
The purpose of thestudies carried out in this periodwas to evaluate the importance of apoLp and LTP in lipid transport and oogenesis. To this end, we reduced the expression (synthesis) of apoLp and LTP genes using RNA interference. The results clearly showed that low expression of apoLp or LTP prevents proper lipid transport to the ovaries and, thus, prevents the formation of eggs and/or the production of fertile eggs. This information may be of importancefrom a general biological perspective because it shows the significance of the two genes studied in the reproduction of this mosquito (Aedes aegypti). However, it may also have some importance for thedesignof newproducts usefulto decrease mosquito populations.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2019
Citation:
Manduca sexta Perilipin 1B: A new PLIN1 isoform linked to fat storage prior to pupation.Insect Biochemistry and Molecular Biology (2019) 110, 69-79.
Xiao Chen, Sarah J Firdaus, Zhiyan Fu, Zengying Wu, Jose L Soulages, Estela L Arrese
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Progress 10/01/17 to 09/30/18
Outputs
Target Audience:The audience of our studies is constituted by biochemists, biologists, and entomologists that areinterested in the metabolism of fat invertebrates and in arthropods. The audience of our studies is distributed around the world in research institutions. Ourstudies were cited in more than 250 articles reported during this 2017-1018 period (Information obtained from GoogleScholar).Graduate and undergraduate students in our department also constitute an important audience for our studies. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Three undergraduate students were trained during this period. One of them, Woong Hee Cho, just joined the graduate program of the University of Texas Southwestern Medical Center. A second undergraduate, Alice Chibnall, joined the lab afew months ago. Sheis intensely working on the project, learninghow to dissect mosquitoes to harvest ovaries, hemolymph and fat body and alsoto separate and analyze proteins by electrophoresis and immunoblot, study gene expression by quantitative PCR and a variety of other common lab techniques. Two graduate students worked and were trained in the lab. One for just a few months, during a rotation, but he may still join the lab to complete a Masters in Biochemistry. The second student, Claudio Galvez Sagastume, finished his second year as agraduate student Claudiocompleted his M.S. and now continues in our lab and graduate program with the intention of earning a Ph.D. degree. How have the results been disseminated to communities of interest?We have collaborated in a study that was published and one of our students reported several results in his thesis. What do you plan to do during the next reporting period to accomplish the goals?As planned, we will continue to study the roles of some genes involved in the metabolism of fat in insects. We will submit some of the completed studies for publication in specialized journals. We will focus on the roles of the lipid droplet specific proteins (PLIN1 and PLIN2) in the accumulationof lipids in fat body and ovaries
Impacts
What was accomplished under these goals?
We have completed the studies on the roles of lipophorin, the mainfat transporter found in the hemolymph of mosquitoes. We have observed that the transport of fat from the place of storage (fat body) to the ovaries is severely limited if the transporter is not properly synthesized in the fat body. The apolipophorin gene was successfully silenced using dsRNA as RNA interference agent. This silencing leads to lipid and protein accumulation in fat body and to a decrease in the lipid content of hemolymph and ovaries. For a successful vitellogenesis, the process of ovary maturation and the formation of viable eggs, Aedes aegypti female mosquitoes must first take a blood meal. Demonstrating a major role of the main lipoprotein of mosquito in ovary maturation and reproduction, apolipophorin silencing affected the size and shape of the ovaries, which remained undeveloped after blood feeding. This result showed that apolipophorin gene expression is essential to the reproduction of Aedes aegypti mosquitoes. Silencing of apolipophorin gene also promoted changes affecting the expression of other lipid transport-related genes at the mRNA and/or protein levels.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2018
Citation:
DmCatD, a cathepsin D-like peptidase of the hematophagous insect Dipetalogaster maxima (Hemiptera: Reduviidae): Purification, bioinformatic analyses and the significance of its interaction with lipophorin in the internalization by developing oocytes. (2018) Jimena Leyria, Leonardo L Fruttero, Rodrigo Ligabue-Braun, Marina S Defferrari, Estela L Arrese, Jos� L Soulages, Beatriz P Settembrini, Celia R Carlini, Lili�n E Canavoso.
Journal of Insect Physiology, 105, 28-39.
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Progress 10/01/16 to 09/30/17
Outputs
Target Audience:The audience of our studies is constituted by biochemists, biologistsand entomologists that areinterested in the metabolism of fat in vertebrates and in arthropods. The audience of our studies is distributed around the world in research institutions. Ourstudies were cited in more than 250 articles reported during this 2016-1017 period (Information obtained from GoogleScholar). Graduate and undergraduate students of our department also constitute an important audience of our studies. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Four undergraduate students were trained during this period. The students learned how to maintain a colony of mosquitoes, how to dissect mosquitoes to harvest ovaries, hemolymph and fat body. They also learned how to separate and analyze proteins by electrophoresis and immunoblot, study gene expression by quantitative PCR and a variety of other common lab techniques. Two graduate students worked and were trained in the lab. One graduate student completed his Ph.D. (Fredy Saudale). The othergraduate student, Claudio Galvez Sagastume, is pursuing a M.S.He has completed the first year of training and course work. How have the results been disseminated to communities of interest?Some of the studies were disseminated to the scientific community in the form of a peer-reviewed publication. Other studies already completed were presented at an international meeting. What do you plan to do during the next reporting period to accomplish the goals?We will continue to study the roles of genes involved in the metabolism of fat in insects. We will put emphasis on the study of the roles ofcertain genes in the fertility of mosquitoes.
Impacts
What was accomplished under these goals?
We have made progress in the understanding of the role of PLIN2 (aim 3) in the regulation of TG storage. We have learned that PLIN2 expression is needed when there is an active synthesis of TG disregard of whether the tissue is accumulating or using and losing TG. Previous studies in Drosophila led to the notion that PLIN2 would help in the accumulation of TG reserves by preventing access of the lipases to the surface of lipid droplets and, thus, also peventing the hydrolysis and mobilization of TG. However, we have shown that the highest expression levelof PLIN2 in fat body is observed when the tissue is in the highest lipolytic state. Our results (recently published) suggest that PLIN2 does not prevent the hydrolysis of TG. It seems like PLIN2 is needed in the assembly of small, nascent, lipid droplets.
Publications
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Progress 10/01/15 to 09/30/16
Outputs
Target Audience:The audience of our studies is constituted by entomologists and scientists in general interested in the metabolism of fat in vertebrates and in arthropods. The audience of our studies is distributed around the world in research institutions. Our studies were cited in more than 200 articles reported during this 2015-1016 period (Information obtained from Google Scholar). Graduate and undergraduate students of our department also constitute an important audience of our studies. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Four undergraduate students were trained during this period. Three of them continue working in the lab. One graduate studentcompleted his Ph.D. (Xiao Chen). We are currently training two graduate students. How have the results been disseminated to communities of interest?Some of the results obtained were disseminated to the scientific community in the form of a peer-reviewed publication. Otherstudies already completed were presented in two international meetings. What do you plan to do during the next reporting period to accomplish the goals?We continue to study the roles of the lipid droplet proteins, PLIN1 and PLIN2, and several other proteinsin lipid metabolism.
Impacts
What was accomplished under these goals?
As indicated in the published article, we have made a majir contribution to the annotation of the genome of the Tobacco hornworm (Manduca sexta). Our specific role was the identification and correction of many of thegenes involved in lipid metabolism. Our work led to the identification of genes of lipid binding proteins, acyltransferases, lipases, lipid droplet proteins and other relevant proteins involved in the synthesis or in the degradation of fat in insects.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2016
Citation:
Multifaceted Biological Insights from a Draft Genome Sequence of the Tobacco Hornworm moth, Manduca sexta
Michael R. Kanost*, Estela L. Arrese, Xiaolong Cao, Yun-Ru Chen, Sanjay Chellapilla, Marian Goldsmith, Ewald Grosse-Wilde, David G. Heckel, Niolae Herndon, Haobo Jiang, Alexie Papanicolaou, Jiaxin Qu, Jose L. Soulages, Heiko Vogel, James Walters, Robert M. Waterhouse,et al.
(2016) Insect Biochem. 76, 118-147
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