Role of cell-secreted vesicles in ovarian follicular fluid during oocyte maturation and their involvement in controlling epigenetic changes

• Grant number: 13/10473-3
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: September 01, 2013
• End date: August 31, 2015
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Flávio Vieira Meirelles
• Grantee: Juliano Coelho da Silveira
• Host Institution: Faculdade de Zootecnia e Engenharia de Alimentos (FZEA). Universidade de São Paulo (USP). Pirassununga , SP, Brazil
• Associated research grant: 12/50533-2 - GIFT: genomic improvement of fertilization traits in Danish and Brazilian cattle, AP.TEM

Abstract

The use of assisted reproduction techniques presents an alternative to improve fertility and genetic selection of domestic species with economic value. However, there is concern that epigenetic changes occurring during in vitro culture or following Intra-cytoplasmic Sperm Injection adversely affect tissue programming during fetal development and could lead to a higher pre-disposition to develop reproductive problems, in adulthood. Epigenetic changes are regulated by methylation and by microRNAs. MicroRNAs are non-coding RNA molecules that regulate gene expression during cell differentiation and tissue tissue development. Recently, cell-secreted vesicles called exosomes and microvesicles carrying bioactive materials including proteins and miRNAs were identified in different body fluids, including ovarian follicular fluid. Cell-secreted vesicles are considered a new class of intercellular communication factors with important roles in different physiological processes. The hypothesis that will be tested is that cell-secreted vesicles containing protein and miRNAs are capable of regulating epigenetic changes during in vitro culture of gametes and embryos. In order to test this hypothesis, ovarian follicle (3-4 mm) will be isolated and oocytes in vitro matured. Oocytes will be assigned to three different experimental groups according to their maturation and fertilization competence. In a retrospective experiment, cell-secreted vesicles will be isolated from follicular fluid according to the oocyte groups. The first aim is to determine protein and total RNA (mRNA, miRNA and snoRNA) contents of microvesicles and exosomes isolated from follicular fluid according to oocyte competence. The second aim is to determine the cellular origin of cell-secreted vesicles isolated from follicular fluid based on protein and RNA markers. The third aim is to evaluate the role of microvesicles and exosomes modulating epigenetic changes during oocyte maturation and embryo development. The results obtained in this research will establish a role for cell-secreted vesicles originating from follicular cells on regulation of epigenetic changes acquired during in vitro culture of oocytes and embryos, generating new therapeutic tools for domestic agriculture species and humans.

Scientific publications

(4)

(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)

DA SILVEIRA, JULIANO C.; DE ANDRADE, GABRIELLA M.; NOGUEIRA, MARCELO F. G.; MEIRELLES, FLAVIO V.; PERECIN, FELIPE. Involvement of miRNAs and Cell-Secreted Vesicles in Mammalian Ovarian Antral Follicle Development. REPRODUCTIVE SCIENCES, v. 22, n. 12, p. 1474-1483, DEC 2015. (13/10473-3, 12/50533-2, 13/08135-2)

ANDRADE, G. M.; MEIRELLES, F. V.; PERECIN, F.; DA SILVEIRA, J. C.. Cellular and extracellular vesicular origins of miRNAs within the bovine ovarian follicle. REPRODUCTION IN DOMESTIC ANIMALS, v. 52, n. 6, p. 1036-1045, DEC 2017. (13/10473-3, 13/08135-2, 12/50533-2, 14/22887-0, 14/21034-3)

DA SILVEIRA, JULIANO C.; DE AVILA, ANA CLARA F. C. M.; GARRETT, HANNAH L.; BRUEMMER, JASON E.; WINGER, QUINTON A.; BOUMA, GERRIT J.. Cell-secreted vesicles containing microRNAs as regulators of gamete maturation. Journal of Endocrinology, v. 236, n. 1, p. R15-R27, JAN 2018. (13/10473-3, 15/16985-1, 12/50533-2, 17/02037-0, 14/22887-0)

DA SILVEIRA, JULIANO C.; ANDRADE, GABRIELLA M.; DEL COLLADO, MAITE; SAMPAIO, RAFAEL V.; SANGALLI, JULIANO R.; SILVA, LUCIANO A.; PINAFFI, FABIO V. L.; JARDIM, IZABELLE B.; CESAR, MARCELO C.; NOGUEIRA, MARCELO F. G.; et al. Supplementation with small-extracellular vesicles from ovarian follicular fluid during in vitro production modulates bovine embryo development. PLoS One, v. 12, n. 6, JUN 15 2017. (13/10473-3, 13/08135-2, 12/50533-2, 14/22887-0, 14/21034-3)