The role of extracellular vesicles in the intercellular communication within the bovine cumulus-oocyte complex during in vitro maturation

• Grant number: 18/14869-2
• Support Opportunities: Scholarships abroad - Research Internship - Master's degree
• Start date: October 31, 2018
• End date: February 26, 2019
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Juliano Coelho da Silveira
• Grantee: Ana Clara Faquineli Cavalcante Mendes de Ávila
• Supervisor: Claude Robert
• Host Institution: Faculdade de Zootecnia e Engenharia de Alimentos (FZEA). Universidade de São Paulo (USP). Pirassununga , SP, Brazil
• Institution abroad: Université Laval, Canada
• Associated to the scholarship: 17/02037-0 - Description of a new mechanism involved in cellular communication between female reproductive system cells and oocyte mediated by extracellular vesicles and their impacts on oocyte maturation, fertilization and bovine embryo cleavage, BP.MS

Abstract

Assisted reproductive techniques (ART) are alternatives to infertility and to increase the production of domestic animals. Although these techniques are widely used, they can cause negative consequences since early pregnancy until adult life. Attempts to mimic in vivo environment are applied to improve problems caused by ARTs. Extracellular vesicles (EVs) secreted by cells are important targets due to their function in intercellular communication. These EVs are present in different body fluids and have bioactive material, like RNAs and proteins. EVs were demonstrated to play a role during in vitro oocyte maturation and embryo development. However, the mechanism of communication mediated by EVs and cumulus-oocyte complex (COCs) still not clear. Then, the hypothesis of this study is that small EVs secreted by follicular fluid can be uptake by cumulus cell and transported to ooplasm by transzonal projections (TZPs) influencing RNA accumulation within the oocyte. Our goal is to investigate the relationship between small EVs from follicular fluid and COCs during in vitro maturation as well as the impact of this transit with RNA accumulation within the oocytes. For this, EVs will be isolated from bovine follicular fluid and stained with a membrane dye and supplemented to maturation media in order to determine the intracellular transit by live imaging. Additionally, COCs supplemented with small EVs will be collected in 0, 2 and 4 hours of in vitro maturation. Total RNA and actin filaments will be stained to quantify RNA within TZPs and ooplasm using confocal microscopy. In conclusion, this study will describe an important mechanism of intercellular communication mediated by EVs during oocyte maturation and its influence in RNA transport. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

CAVALCANTE MENDES DE AVILA, ANA CLARA FAQUINELI; BRIDI, ALESSANDRA; ANDRADE, GABRIELLA MAMEDE; DEL COLLADO, MAITE; SANGALLI, JULIANO RODRIGUES; NOCITI, RICARDO PERECIN; DA SILVA JUNIOR, WILSON ARAUJO; BASTIEN, ALEXANDRE; ROBERT, CLAUDE; MEIRELLES, FLAVIO VIEIRA; et al. Estrous cycle impacts microRNA content in extracellular vesicles that modulate bovine cumulus cell transcripts during in vitro maturation. BIOLOGY OF REPRODUCTION, v. 102, n. 2, p. 362-375, FEB 2020. (14/22887-0, 18/14869-2, 17/02037-0, 15/21829-9)