Lipid and amino acid metabolism during bovine oocyte in vitro maturation.

MARTINEZ SANTIAGO; CETICA PABLO; GAGNETEN PAULA; GUTNISKY CYNTHIA; BREININGER ELIZABETH

CABA

Jornada; Jornadas Internacionales del INITRA; 2021

INITRA- FAcultad de Ciencias Veterinarias, UBA

Most metabolic studies carried out in cumulus oocytes-complexes (COCs) refer to carbohydrates as they are the main substrates in the culture media. There is limited information about the use of endogen lipids (EL) or amino acid (AA) as oxidative substrates. The aim of this work was to evaluate the endogen lipid consumption and amino acid utilization by COCs during bovine oocyte maturation in a media lacking other oxidative substrates. Maturation was performed in a defined media (SOFm -without pyruvate and lactate supplemented with FSH, LH, EGF, insulin, gentamicin and PVA) to which were added different supplementations. For the EL study COCs were randomly divided in 5 groups: a) without supplementation, b) glucose, c) L-carnitine (β-oxidation fatty acid stimulator), d) L-carnitine + glucose y e) etomoxir (β-oxidation fatty acid inhibitor). For the AA as oxidative substrates study COCs were randomly divided in 5 groups: a) without supplementation b) glucose, c) AA, d) AA + glucose y e) AA + salicilate (glutamate dehydrogenase inhibitor). The proportion of nuclear maturation was evaluated by the presence of metaphase II after Hoechst 33342 staining. The COCs EL content was determined by Nile Red staining (n=35-45 per group). For the AA study the ammonia concentration of the media after maturation was measured using a spectrophotometric assay (n=8-10 per group). The proportion of nuclear maturation was compared using a Chi-squared test for non-parametric data (n=65-99 per group). The EL content and ammonia production were compared by analysis of variance (ANOVA). P