Digestion and midgut absorption in 3 insects:an enzymological, transcriptomic , and proteomic approach.

• Grant number: 19/21065-0
• Support Opportunities: Research Grants - Visiting Researcher Grant - Brazil
• Duration: February 17, 2020 - February 16, 2021
• Field of knowledge: Biological Sciences - Biochemistry
• Principal Investigator: Walter Ribeiro Terra
• Grantee: Walter Ribeiro Terra
• Visiting researcher: Carlos Peres Silva
• Visiting researcher institution: Universidade Federal de Santa Catarina (UFSC). Centro de Ciências Biológicas, Brazil
• Host Institution: Instituto de Química (IQ). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 17/08103-4 - Insect digestion: a molecular, cellular, physiological and evolutionary approach, AP.TEM

Abstract

A project for a visiting researcher for one year at the University of São Paulo, Invited research: Dr. Carlos Peres SilvaProject: Digestion and midgut absorption in 3 insects. An enzymological, transriptomic and proteomic approach.This project is part of the Projeto Tematico FAPESP (Process: 2017/08103-4), coordinated by Dr. Walter R. Terra, which intends to describe the molecular processes underlying digestion and absorption in insects pertaining to different orders.The specific proposal of this investigation is to identify the coding genes for the main digestive enzymes reported for Zabrotes subfasciatus (Silva et al. Insect Biochem. Mol. Biol. 11999) 29: 355-366) and Dermestes maculatus (Caldeira et al., Archs. Insect Biochem. Physiol. (2007) 64: 1-19), as well as of the chief ion and nutrient transporters involved in midgut absorption. The procedures to be used regarding digestive enzymes will be the same we employed for Periplaneta americana (Tamaki et al., J. Insect Physiol. (2014) 70: 22-35) and those referring to transporters as we accomplished with Tenebrio molitor (Moreira et al., J. Insect Physiol. (2017) 99: 58-66) and Musca domestica (Pimentel tet al., J. Insect Physiol. (2018) 109: 11-20; Barroso et al., Comp. Biochem. Physiol. Part A (2019) 337, 110535). The functional testes designed to test the proposed models will be inspired in those used in Musca domestica.Another branch of the project is the identification by proteomics of the transporters present in midgut microvillar and perimicrovillar membranes of Rhodnius prolixus (Terra et al., Insect Biochem. (1988) !8: 423-434; Ferreira et al., Insect Biochem.(1988) 18: 521-530). The perimicrovillar membranes are membranes that ensheath the microvillar membranes as glove fingers in hemipterans like R. prolixus. The experimental challenge here is to isolate the membranes in masses enough for proteomics analyses.The laboratoty has transcriptomes in triplicate of several midgut regions of the 3 insects and a bioinformatic technician to support part of the project. The preparation of the midgut microvillar and perimicrovillar membranes will be done in collaboration with Dr. Fernando A. Genta da FIOCRUZ-RF and Dr. Ignacio G. Barroso of our laboratory. The proteomic analyses will be done in the facilities of the University of São Paulo.The schedule to be followed will be the following:1. Search for ion and nutrient transporters and digestive enzymes in Z. subfasciatus and D. maculatus.2. Expression analyses ot transporters and enzymes along the midgut of the insects.3. Proposals of midgut molecular mechanisms in insects and their corresponding functional roles.4. Proteomics of the midgut microvillar and perimicrovillar membranes of R. prolixus and proposals of their participation in midgut physiology.The collaboration of Dr. Carlos P. Silva will be important for the success of the project, because he has experience with Z. subfasciatus biology (Silva et al., Insect Biochem. Mol. Biol. (1999) 29: 355-366), which is crucial in this project and that will help him to deal with another beetle, D. maculatus. Dr. Silva also has experience in isolating midgut microvillar and perimicrovillar membranes of hemipterans (Silva et al., Insect Biochem. Mol. Biolo. (1996) 26 : 1011-1118. (AU)