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Differential gene expression, ultrastructural analysis and evaluation of the lipid profile of in vitro produced bovine blastocysts from oocytes matured conventionally or via SPOM system

Grant number: 12/23409-9
Support type:Scholarships in Brazil - Doctorate
Effective date (Start): March 01, 2013
Effective date (End): February 10, 2017
Field of knowledge:Agronomical Sciences - Veterinary Medicine
Principal Investigator:Marcelo Fábio Gouveia Nogueira
Grantee:Eduardo Montanari Razza
Home Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil
Associated research grant:12/50533-2 - GIFT: genomic improvement of fertilization traits in Danish and Brazilian cattle, AP.TEM
Associated scholarship(s):15/02557-8 - Ultrastructure analysis of bovine oocytes and blastocysts produced in vitro after oocyte maturation with cAMP modulators, BE.EP.DR

Abstract

In vitro maturation (IVM) do not use superestimulatory hormones, which avoid excessive spending and manipulations of the animal and also the harmful iatrogenic effects of ovarian overstimulation. However, the efficiency of IVM does not meet expectations due to its inability to simulate the physiological events that occur in vivo (as the main example, the synchronization between the nuclear and cytoplasmic maturation of the oocyte). For this reason, the increment of IVM media is a target sought increasingly in reproductive studies. The evaluation of the lipid profile of embryos is a new and particularly important approach for gauging the embryo ability for cryopreservation. In addition, cell characterization through assessment of morphology and organelles distribution, provides consistent results on embryo characterization when examined ultrastructurally. Furthermore, the differential expression of genes - known as indicators of specific function and hence embryonic competence - can be evaluated in combination with such techniques in order to identify in vitro the blastocysts that have better quality (morphological and physiological) and competence for later implantation and development. Therefore, this study aims to compare the conventional IVM system with the new system simulated physiology oocyte maturation - SPOM), which uses drugs and control the specific timing of their administration (briefly: forskolin and IBMX on pre-maturation for 2 h, and cilostamide and FSH for the next 28 h of maturation). Oocytes will be matured, fertilized and the produced blastocysts will be evaluated for the lipid profile (mass spectrometry MALDI-TOF-MS, Thomson Laboratory, Campinas), the ultrastructure analysis (electron microscopy, Institute of Biosciences, Botucatu) and differential expression of 45 genes indicators of quality and competence of embryonic development (microfluidic system, Fluidigm). The methodology described will be held in Brazil (Institute of Biosciences, Botucatu), with Bos indicus cattle, and in Denmark (Department of Basic Animal and Veterinary Sciences, University of Copenhagen) with Bos taurus cattle in order to verify the efficiency and repeatability of the SPOM system with different scenarios and genotypes. The communion of the data to be obtained with this project aims to provide a general and reliable marker of embryo quality, covering the implantation, development and cryopreservation of embryos in commercial routines of IVP.

Scientific publications (5)
(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.; PEDERSEN, HANNE S.; STROEBECH, LOTTE; FONTES, PATRICIA K.; KADARMIDEEN, HAJA N.; CALLESEN, HENRIK; PIHL, MARIA; NOGUEIRA, MARCELO F. G.; HYTTEL, POUL. Simulated physiological oocyte maturation has side effects on bovine oocytes and embryos. JOURNAL OF ASSISTED REPRODUCTION AND GENETICS, v. 36, n. 3, p. 413-424, MAR 2019. Web of Science Citations: 1.
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.
BOTIGELLI, RAMON CESAR; RAZZA, EDUARDO MONTANARI; PIOLTINE, ELISA MARIANO; FONTES, PATRICIA KUBO; LANCELLOTTI SCHWARZ, KATIA REGINA; VERDE LEAL, CLAUDIA LIMA; GOUVEIA NOGUEIRA, MARCELO FABIO. Supplementing in vitro embryo production media by NPPC and sildenafil affect the cytoplasmic lipid content and gene expression of bovine cumulus-oocyte complexes and embryos. REPRODUCTIVE BIOLOGY, v. 18, n. 1, p. 66-75, MAR 2018. Web of Science Citations: 1.
BOTIGELLI, RAMON CESAR; RAZZA, EDUARDO MONTANARI; PIOLTINE, ELISA MARIANO; GOUVEIA NOGUEIRA, MARCELO FABIO. New approaches regarding the in vitro maturation of oocytes: manipulating cyclic nucleotides and their partners in crime. JORNAL BRASILEIRO DE REPRODUCAO ASSISTIDA, v. 21, n. 1, p. 35-44, JAN-MAR 2017. Web of Science Citations: 4.
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: 5.
Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
RAZZA, Eduardo Montanari. Expressão gênica diferencial, análise ultraestrutural e avaliação do perfil lipídico de blastocistos bovinos produzidos in vitro a partir de oócitos maturados convencionalmente ou pelo sistema SPOM. 2017. Doctoral Thesis - Universidade Estadual Paulista "Júlio de Mesquita Filho" Instituto de Biociências (Campus de Botucatu)..

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