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Intra and intercellular routes during foliculogenesis and in vivo and in vitro oocyte maturation

Grant number: 14/21034-3
Support type:Regular Research Grants
Duration: March 01, 2015 - February 28, 2017
Field of knowledge:Agronomical Sciences - Veterinary Medicine
Principal Investigator:Felipe Perecin
Grantee:Felipe Perecin
Home Institution: Faculdade de Zootecnia e Engenharia de Alimentos (FZEA). Universidade de São Paulo (USP). Pirassununga , SP, Brazil
Assoc. researchers:Fabiana Fernandes Bressan ; Juliano Coelho da Silveira ; Luciano Andrade Silva

Abstract

In vitro production (IVP) of bovine embryos is a widely used biotechnology in Brazil. However, the in vitro embryo production system, although technically viable, leads to cellular and molecular alteration in oocytes and embryos, producing undesirable outcomes and embryo developmental rates ranging from 30% to 50%. Therefore, generate methodologies the lead to higher conversion of oocytes to embryos, and comprehend the mechanism that determine the oocyte competence is strategic for the Brazilian industry of embryo in vitro production. The follicular microenvironment and the oocyte metabolism are fundamental to determine the competence acquisition of the oocyte. Experiments evaluating intra and intercellular routes during the oocyte maturation and embryo development will be conducted aiming to investigate the biological process associated with the acquisition of competence and to generate increments on embryo in vitro production rates. The first research strategy will focus on lipid oxidative metabolism and mitochondrial activity. Previous studies conducted by our group showed a deficient migration and total quantity of active mitochondria and lipid droplets in in vitro maturation (IVM) compared to in vivo matured oocytes. Since that oocytes' mitochondria are responsible for the energetic supply until the blastocyst stage, and that fatty acids oxidation is the main cellular mechanism of energetic supply for oocyte and embryo development, the activity of both mitochondria and lipid droplets are fundamental for IVP. Previous reports show that modulator of beta-oxidation, such as L-carnitine and bezafibrate, can stimulate the oxidative process and lead to an increase of in vitro embryo production rates. Thus, this line of investigation aim to compare the mitochondrial activity and the lipid metabolism between in vivo and in vitro embryo production, as well as assess the effect of oxidative modulators supplementation (L-carnitine and bezafibrate) during IVM on embryo IVP rates. Both the lipid droplets and mitochondrial behavior will be evaluated, as well as the cellular oxidative stress during in vivo and in vitro maturation and development. The second and third research strategies will focus on extracellular routs of communication within the follicular microenvironment. The hypothesis that will be investigated is that proteins enclosed on cell-secreted vesicles (CSV) in the follicular fluid (FF) and miRNAs from cumulus cells modulate oocyte quality. Proteins in CSV (microvesicles and exosomes) from the FF associated with different developmental competences will be identified and quantified; and miRNA profile of cumulus cell correlated with oocyte competence will be determined. To validate miRNAs associated with oocyte development competence, functional tests will be performed mimicking (miRNA mimics) or inhibiting (miRNA inhibitors) target miRNAs previously related to the presence of absence of oocyte competence, and the effects on embryo IVP will be assessed. Results of these experiments will help to elucidate the effects of in vitro systems on beta-oxidation and lipid-mitochondrial metabolism, as well as help to clarify the mechanisms of intercellular communication within the ovarian follicle, characterize miRNA profile of cumulus cells, and identify potential miRNAs and protein biomarkers for oocyte quality. Moreover, the results will enable the development of new tools to improve embryo IVP through supplementation of modulators of mitochondrial oxidation that prevent cellular oxidative stress or through the mimicking of miRNA profiles associated with oocyte competence. (AU)

Scientific publications (10)
(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)
ANDRADE, GABRIELLA MAMEDE; BOMFIM, MONALISA MEDRADO; DEL COLLADO, MAITE; MEIRELLES, FLAVIO VIEIRA; PERECIN, FELIPE; DA SILVEIRA, JULIANO COELHO. Oxygen tension modulates extracellular vesicles and its miRNA contents in bovine embryo culture medium. Molecular Reproduction and Development, v. 86, n. 8, p. 1067-1080, AUG 2019. Web of Science Citations: 0.
SANGALLI, JULIANO RODRIGUES; SAMPAIO, RAFAEL VILAR; DEL COLLADO, MAITE; DA SILVEIRA, JULIANO COELHO; CAMARA DE BEM, TIAGO HENRIQUE; PERECIN, FELIPE; SMITH, LAWRENCE CHARLES; MEIRELLES, FLAVIO VIEIRA. Metabolic gene expression and epigenetic effects of the ketone body beta-hydroxybutyrate. SCIENTIFIC REPORTS, v. 8, SEP 13 2018. Web of Science Citations: 2.
DEL COLLADO, MAITE; ANDRADE, GABRIELLA MAMEDE; MEIRELLES, FLAVIO VIEIRA; DA SILVEIRA, JULIANO COELHO; PERECIN, FELIPE. Contributions from the ovarian follicular environment to oocyte function. ANIMAL REPRODUCTION, v. 15, n. 3, p. 261-270, JUL-SEP 2018. Web of Science Citations: 1.
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. Web of Science Citations: 8.
BOMFIM, MONALISA M.; ANDRADE, GABRIELLA M.; DEL COLLADO, MAITE; SANGALLI, JULIANO R.; FONTES, PATRICIA K.; NOGUEIRA, MARCELO F. G.; MEIRELLES, FLAVIO V.; DA SILVEIRA, JULIANO C.; PERECIN, FELIPE. Antioxidant responses and deregulation of epigenetic writers and erasers link oxidative stress and DNA methylation in bovine blastocysts. Molecular Reproduction and Development, v. 84, n. 12, p. 1296-1305, DEC 2017. Web of Science Citations: 5.
DEL COLLADO, MAITE; DA SILVEIRA, JULIANO C.; OLIVEIRA, MARCELO L. F.; ALVES, BARBARA M. S. M.; SIMAS, ROSINEIDE C.; GODOY, ADRIANA T.; COELHO, MIRELA B.; MARQUES, LYGIA A.; CARRIERO, MATEUS M.; NOGUEIRA, MARCELO F. G.; EBERLIN, MARCOS N.; SILVA, LUCIANO A.; MEIRELLES, FLAVIO V.; PERECIN, FELIPE. In vitro maturation impacts cumulus-oocyte complex metabolism and stress in cattle. Reproduction, v. 154, n. 6, p. 881-893, DEC 2017. Web of Science Citations: 3.
ANDRADE, GABRIELLA MAMEDE; DA SILVEIRA, JULIANO COELHO; PERRINI, CLAUDIA; DEL COLLADO, MAITE; GEBREMEDHN, SAMUEL; TESFAYE, DAWIT; MEIRELLES, FLAVIO VIEIRA; PERECIN, FELIPE. The role of the PI3K-Akt signaling pathway in the developmental competence of bovine oocytes. PLoS One, v. 12, n. 9 SEP 18 2017. Web of Science Citations: 10.
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.; CESAR, ALINE S. M.; COUTINHO, LUIZ L.; PEREIRA, RINALDO W.; PERECIN, FELIPE; MEIRELLES, FLAVIO V. 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. Web of Science Citations: 11.
DEL COLLADO, MAITE; DA SILVEIRA, JULIANO COELHO; SANGALLI, JULIANO RODRIGUES; ANDRADE, GABRIELLA MAMEDE; DA SILVA SOUSA, LETICIA RABELLO; SILVA, LUCIANO ANDRADE; MEIRELLES, FLAVIO VIEIRA; PERECIN, FELIPE. Fatty Acid Binding Protein 3 And Transzonal Projections Are Involved In Lipid Accumulation During In Vitro Maturation Of Bovine Oocytes. SCIENTIFIC REPORTS, v. 7, JUN 1 2017. Web of Science Citations: 13.
BRESSAN, FABIANA FERNANDES; FANTINATO-NETO, PAULO; ANDRADE, GABRIELLA MAMEDE; SANGALLI, JULIANO RODRIGUES; SAMPAIO, RAFAEL VILAR; DA SILVEIRA, JULIANO COELHO; PERECIN, FELIPE; MEIRELLES, FLAVIO VIEIRA. Challenges and perspectives to enhance cattle production via in vitro techniques: focus on epigenetics and cell-secreted vesicles. Ciência Rural, v. 45, n. 10, p. 1879-1886, OCT 2015. Web of Science Citations: 1.

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