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Identifier 000443928
Title In silico and in vivo characterization of the ecdysone importer Oatp74D in Lepidoptera
Alternative Title In silico and in vivo ανάλυση το μεταφορέα ενδυσόνης Oatp74D στα λεπτιδόπτερα
Author Φωτιάδου, Μελίνα Χ.
Thesis advisor Βόντας, Ιωάννης
Reviewer Παυλίδης, Παύλος
Siden-Kiamos, Inga
Abstract Insect pests damage agricultural production to a great extent, imposing significant costs to global economy. At the same time, the intensive use of insecticides often brings about populations resistant to the existing insecticides. At present, two major lepidopteran pests are Helicoverpa armigera and Spodoptera frugiperda. These species are considered to be so destructive, because they are polyphagous, invasive, widely distributed and have already developed resistance to existing insecticides via target-site mutations. Thus, the discovery of new targets for drug development, in order to control their populations, is of utmost importance. Recent research in the model insect Drosophila melanogaster reports the role of a plasma membrane transporter referred as Ecdysone Importer (EcI), also known as organic anion transporting polypeptide 74D (Oatp74D), in the development of D. melanogaster. Ecdysone is a steroid hormone that regulates several physiological processes. When ecdysone enters the cell it binds and activates an intracellular nuclear receptor (EcR) which is then able to induce the transcription of several genes essential for the development of the organism. It was shown that EcI is necessary for ecdysone signaling. This transporter belongs to a specific subclade of the Solute Carrier O (SLCO) family of transporters, which is well represented in insects while being absent in mammals. These two characteristics of EcI, the essentiality for the development of the organism and the insect specificity, render it a potential target for the development of insecticides. However, further analysis of this transporter is needed in pest species to prove whether it is appropriate as a drug target. In this study, we constructed a phylogenetic tree of the SLCO transporters of several ecdysozoa, organisms that utilize ecdysosteroids. The gene family tree provided information on which species have potential orthologs of Oatp74D and showed that this gene probably appeared before the evolution of arthropods, but after the evolution of ecdysozoa. After finding the potential orthologs of Oatp74D in the pests of our interest, we proceeded to functional characterization of the lepidopteran Oatp74D. By making use of CRISPR Cas9 system we managed to generate mosaic knockouts of Oatp74D in S. frugiperda embryos and found that this gene is necessary for the development of the organism at embryonic and larval stages.
Language English
Subject Frugiperda
Issue date 2021-11-26
Collection   School/Department--School of Sciences and Engineering--Department of Biology--Post-graduate theses
  Type of Work--Post-graduate theses
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