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Effect of the modulation of metabolic pathways on the acetylation pattern of bovine oocytes histones during in vitro maturation

Grant number: 18/23142-9
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): February 01, 2019
Effective date (End): March 31, 2021
Field of knowledge:Agronomical Sciences - Veterinary Medicine - Animal Reproduction
Principal Investigator:Marcella Pecora Milazzotto
Grantee:João Vitor Alcantara da Silva
Home Institution: Centro de Ciências Naturais e Humanas (CCNH). Universidade Federal do ABC (UFABC). Ministério da Educação (Brasil). Santo André , SP, Brazil

Abstract

In vitro maturation (MIV) is one of the primordial steps for the success of in vitro production (PIV) of bovine Embrys, since there is the triggering of severe processes that collaborate for the development and the quality of the future embryo. Epigenetic reprogramming is one of the processes carried out in MIV, in which the bovine oocyte goes through an intense regulation of Epigenetic mechanisms, mainly the acetylation of the histones, with the aim of acquiring Oocitária competence. In addition, the via glycolytic, performed in the cumulus cells, and the tricarboxylic acid cycle (TCA) are the main pathways that provide acetyl and energy groups for the bovine oocyte. However, little is known about the import of energy metabolism, by the availability of acetyl-coenzyme A (acetyl-Coa), in the mechanisms of histones acetylation and in obtaining Oocitária competence. With This, the objective of this work is to analyze and investigate the effects of the pharmacological modulation of the main metabolic pathways in the acetylation pattern of the histones of oocytes bovine animals in MIV. The methodology shall consist in the acquisition of the oocytes and realization of the MIV in the presence or absence of the inhibitor of the via glycolytic, the iodoacetate, and the stimulator of the TCA cycle, the dichloroacetic acid. The oocytes will be analyzed by immunofluorescence for the acetylation of lysine 9 of histone 3 (H3) and the Cumulus cells by Fluorimetria Kit to observe the global acetylation standard of H3. It is expected that the inhibition of the glycolytic will result in the decrease of acetyl groups, resulting in the decrease of the acetylation in the histones and, consequently, the gene transcription. On the other hand, stimulation of the TCA cycle will result in greater availability of acetyl-CoA, leading to increased acetylation of histones and increase in gene transcription.