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COMPARISON BETWEEN THE CONVENTIONAL IN VITRO MATURATION AND THE SIMULATION OF OOCYTE PHYSIOLOGICAL MATURATION (SPOM) IN BOVINE CUMULUS-OOCYTE COMPLEX

Grant number: 12/10737-8
Support type:Scholarships in Brazil - Post-Doctorate
Effective date (Start): September 01, 2012
Effective date (End): August 31, 2016
Field of knowledge:Agronomical Sciences - Veterinary Medicine - Animal Reproduction
Principal researcher:Marcelo Fábio Gouveia Nogueira
Grantee:Mariana Fernandes Machado
Home Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil
Associated scholarship(s):13/12960-9 - Comparison between conventional and the simulated physiological oocyte maturation (SPOM) systems of maturation in bovine cumulus-oocyte complexes, BE.EP.PD

Abstract

The inefficiency of in vitro maturation (IVM) compared to in vivo maturation is due to spontaneous resumption of meiosis I in the oocyte when removed from the ovarian follicle. The cAMP is a key molecule that regulates oocyte maturation. A decrease in the oocyte cAMP leads to premature resumption of meiosis I and asynchrony between the nuclear and cytoplasmic maturation. Several substances that stimulate the synthesis or inhibiting degradation of cAMP in the oocyte have been used to simulate a physiological maturation and thus improving the in vitro embryos production (IVP). This knowledge led to the development of SPOM system (simulated physiological oocyte maturation), which consists of a prolonged period of maturation and simultaneously delayed nuclear maturation by the addition of substances that maintains the oocyte cAMP in high concentrations. The success of SPOM system in bovine and mice embryo production in vitro motivates further investigations regarding cellular and molecular mechanisms of this system. It is well known that the oocyte I, arrested at the germinal vesicle stage (GV), has high transcriptional activity. Consequently, blocking the maturation at this stage allows the storage of RNAs and proteins that will support the development of the embryo until its genome activation. Furthermore, the conventional maturation systems can modify the molecular mechanisms that regulate the oocyte development. Therefore, the present project aim to identify genes differentially expressed between the conventional IVM system and the SPOM. Cumulus-oocyte complexes, majority zebu obtained from an abattoir, will be matured (conventional and SPOM) and then analyzed regarding global gene expression by microarray technique. After microarray analysis, eight candidate genes will be selected by differential expression between the two systems and validated by real time PCR. The functionality of the SPOM system will be assessed by nuclear oocyte maturation and gap junction quantification in the cumulus cells. At the end of the IVP, blastocysts rates and embryo quality will also be assessed. We believe that the identification of genes differentially expressed, and the relation with their functions may help us to understand the mechanisms regulating oocyte maturation, and further to improve the conditions of IVM and bovine embryos production.

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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)
RAZZA, EDUARDO M.; SUDANO, MATEUS J.; FONTES, PATRICIA K.; FRANCHI, FERNANDA F.; BELAZ, KATIA ROBERTA A.; SANTOS, PRISCILA H.; CASTILHO, ANTHONY C. S.; ROCHA, DANIELE F. O.; EBERLIN, MARCOS N.; MACHADO, MARIANA F.; NOGUEIRA, MARCELO F. G. Treatment with cyclic adenosine monophosphate modulators prior to in vitro maturation alters the lipid composition and transcript profile of bovine cumulus-oocyte complexes and blastocysts. REPRODUCTION FERTILITY AND DEVELOPMENT, v. 30, n. 10, p. 1314-1328, SEP 2018. Web of Science Citations: 2.
ERENO, R. L.; LOUREIRO, B.; CASTILHO, A. C. S.; MACHADO, M. F.; PEGORER, M. F.; SATRAPA, R. A.; NOGUEIRA, M. F. G.; BURATINI, J.; BARROS, C. M. Expression of mRNA Encoding the LH Receptor (LHR) and LHR Binding Protein in Granulosa Cells from Nelore (Bos indicus) Heifers Around Follicle Deviation. REPRODUCTION IN DOMESTIC ANIMALS, v. 50, n. 6, p. 952-957, DEC 2015. Web of Science Citations: 2.
CASTILHO, A. C. S.; NOGUEIRA, M. F. G.; FONTES, P. K.; MACHADO, M. F.; SATRAPA, R. A.; RAZZA, E. M.; BARROS, C. M. Ovarian superstimulation using FSH combined with equine chorionic gonadotropin (eCG) upregulates mRNA-encoding proteins involved with LH receptor intracellular signaling in granulosa cells from Nelore cows. Theriogenology, v. 82, n. 9, p. 1199-1205, DEC 2014. Web of Science Citations: 3.

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