New method to evaluate intercellular communication mediated by extracellular vesicles within the bovine ovarian follicle

• Grant number: 17/24326-3
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: July 01, 2018
• End date: June 30, 2019
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Juliano Coelho da Silveira
• Grantee: Victória Martins Braghetto Barillari
• Host Institution: Faculdade de Zootecnia e Engenharia de Alimentos (FZEA). Universidade de São Paulo (USP). Pirassununga , SP, Brazil
• Associated research grant: 14/22887-0 - Cell-secreted vesicles containing miRNAs modulate epigenetic changes during in vitro culture of bovine gametes and embryos, AP.JP

Abstract

Numerous studies have now been performed to optmize assisted reproduction techniques. The efficiency of the in vitro embryo production is dependent on intercellular communication, and this communication can be mediated by extracellular vesicles (microvesicles and exosomes), which can be secreted and incorporated by different cell types. In this way, new tools to determine the role of extracellular vesicles in intercellular comumunication within the ovarian environment are necessary for the progress of this area of knowledge. This study aims to implement a new type of labeling of extracellular vesicles secreted by bovine ovarian follicle granulosa cells. For this, granulosa cells will be transfected with plasmidscalled palmGFP (coding GP43-GFP protein) and the culture medium pf these cells will be used to isolate cell-secreted extracellular vesicles (containing GFP). Additionally, we will use these extracellular vesicles as treatment for cumulus oocyte complexes during in vitro maturation. Based on that we believe that this new method will allow to determine the endocytosis of extracellullar vesicles in cumulus cells avoiding problems associated to the traditional stains. Therefore, this project aims to improve the tracking of extracellular vesicles and thus generate tools to imrpove the study of intercellular communication mediated by extracellular vesicles within the ovarian follicle.