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Metabolomic profiles of bovine cumulus cells and cumulus-oocyte-complex-conditioned medium during maturation in vitro.

ABSTRACT: Cumulus cells are essential for nutrition of oocytes during maturation. In the absence of cumulus cells during maturation, oocyte developmental competence is severely compromised. In this study, we matured bovine cumulus-oocyte-complexes (COCs) for 8?h, the cumulus cells were removed and denuded oocytes were further matured for 15?h in either the medium conditioned by the initial 8?h culture, or in fresh unconditioned medium. Denuded oocytes that completed maturation in COC-conditioned medium demonstrated better developmental potential than denuded oocytes that completed maturation in standard maturation medium. An inventory was made of the metabolites secreted by COCs into the maturation medium during the first 8?h, from 8 to 23?h, and during an entire 23?h maturation protocol; the metabolomic changes in the cumulus cells during maturation were also investigated. In maturation medium, 173 biochemical components were detected compared to 369 different metabolites in cumulus cells. Significant changes in metabolomic components were evident in maturation medium and in cumulus cells during maturation, with most of the changes related to amino acid, carbohydrate, and lipid metabolism. The importance of two detected biochemicals, creatine and carnitine, for oocyte maturation was further investigated. The presence of carnitine, but not creatine during oocyte in vitro maturation improved the developmental competence of denuded oocytes.

SUBMITTER: Uhde K

PROVIDER: S-EPMC6013446 | biostudies-literature | 2018 Jun

REPOSITORIES: biostudies-literature

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Metabolomic profiles of bovine cumulus cells and cumulus-oocyte-complex-conditioned medium during maturation in vitro.

Uhde Karen K van Tol Helena T A HTA Stout Tom A E TAE Roelen Bernard A J BAJ

Scientific reports 20180621 1

Cumulus cells are essential for nutrition of oocytes during maturation. In the absence of cumulus cells during maturation, oocyte developmental competence is severely compromised. In this study, we matured bovine cumulus-oocyte-complexes (COCs) for 8 h, the cumulus cells were removed and denuded oocytes were further matured for 15 h in either the medium conditioned by the initial 8 h culture, or in fresh unconditioned medium. Denuded oocytes that completed maturation in COC-conditioned medium de ...[more]

PMID: 29930262

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Project description:BackgroundIn the cow, lysophosphatidic acid (LPA) acts as an auto-/paracrine factor, through its receptors LPAR1-4, on oocytes and cumulus cells during in vitro maturation (IVM). The aim of the present work was to determine the effect of LPA during IVM of bovine oocytes on: 1) oocyte maturation; 2) apoptosis of COCs; 3) expression of genes involved in developmental competence and apoptosis in bovine oocytes and subsequent blastocysts; 4) cumulus expansion and expression of genes involved in the ovulatory cascade in cumulus cells; 5) glucose metabolism and expression of genes involved in glucose utilization in cumulus cells; 6) cleavage and blastocyst rates on Day 2 and Day 7 of in vitro culture, respectively.MethodsCumulus-oocyte complexes (COCs) were matured in vitro in the presence or absence of LPA (10(-5) M) for 24 h. Following maturation, we determined: oocyte maturation stage, cumulus expansion, COCs apoptosis and glucose and lactate levels in the maturation medium. Moreover, COCs were either used for gene expression analysis or fertilized in vitro. The embryos were cultured until Day 7 to assess cleavage and blastocyst rates. Oocytes, cumulus cells and blastocysts were used for gene expression analysis.ResultsSupplementation of the maturation medium with LPA enhanced oocyte maturation rates and stimulated the expression of developmental competence-related factors (OCT4, SOX2, IGF2R) in oocytes and subsequent blastocysts. Moreover, LPA reduced the occurrence of apoptosis in COCs and promoted an antiapoptotic balance in the transcription of genes involved in apoptosis (BAX and BCL2) either in oocytes or blastocysts. LPA increased glucose uptake by COCs via augmentation of GLUT1 expression in cumulus cells as well as stimulating lactate production via the enhancement of PFKP expression in cumulus cells. LPA did not affect cumulus expansion as visually assessed, however, it stimulated upstream genes of cumulus expansion cascade, AREG and EREG.ConclusionsSupplementation of the maturation medium with LPA improves oocyte maturation rates, decreases extent of apoptosis in COCs and sustains the expression of developmental competence related factors during oocyte maturation and subsequently affects gene expression profile at the blastocyst stage. We also demonstrate that LPA directs glucose metabolism toward the glycolytic pathway during IVM.

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Metabolomic profiles of bovine cumulus cells and cumulus-oocyte-complex-conditioned medium during maturation in vitro.

Publications

Metabolomic profiles of bovine cumulus cells and cumulus-oocyte-complex-conditioned medium during maturation in vitro.

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