Abstract
BackgroundSupplementation of bovine oocyte-cumulus complexes during in vitro maturation (IVM) with 1 μM of docosahexaenoic acid (DHA), C22:6 n-3 polyunsaturated fatty acid, was reported to improve in vitro embryo development. The objective of this paper was to decipher the mechanisms of DHA action.ResultsTranscriptomic analysis of 1 μM DHA-treated and control cumulus cells after 4 h IVM showed no significant difference in gene expression. MALDI-TOF mass spectrometry analysis of lipid profiles in DHA-treated and control oocytes and cumulus cells after IVM showed variations of only 3 out of 700 molecular species in oocytes and 7 out of 698 species in cumulus cells (p < 0.01).We showed expression of free fatty acid receptor FFAR4 in both oocytes and cumulus cells, this receptor is known to be activated by binding to DHA. FFAR4 protein was localized close to the cellular membrane by immunofluorescence. Functional studies demonstrated that supplementation with FFAR4 agonist TUG-891 (1 μM or 5 μM) during IVM led to an increased blastocyst rate (39.5% ± 4.1%, 41.3% ± 4.1%), similar to DHA 1 μM treatment (39.2% ± 4.1%) as compared to control (25.2% ± 3.6%).FFAR4 activation via TUG-891 led to beneficial effect on oocyte developmental competence and might explain in part similar effects of DHA.ConclusionsIn conclusion, we suggested that low dose of DHA (1 μM) during IVM might activate regulatory mechanisms without evident effect on gene expression and lipid content in oocyte-cumulus complexes, likely through signaling pathways which need to be elucidated in further studies.
Highlights
Supplementation of bovine oocyte-cumulus complexes during in vitro maturation (IVM) with 1 μM of docosahexaenoic acid (DHA), C22:6 n-3 polyunsaturated fatty acid, was reported to improve in vitro embryo development
Effect of 1 μM DHA on gene expression in cumulus cells during IVM In order to decipher the possible mechanism of the beneficial effect of 1 μM DHA on oocyte quality, we compared the transcriptomic profiles of control and DHA-treated cumulus cells after 4 h IVM by microarray
Among the 35 up-regulated genes in DHA treated cumulus cells (CC), 20 genes corresponded to small nucleolar Ribonucleic acid (RNA) and five genes corresponded to spliceosomal RNA
Summary
Supplementation of bovine oocyte-cumulus complexes during in vitro maturation (IVM) with 1 μM of docosahexaenoic acid (DHA), C22:6 n-3 polyunsaturated fatty acid, was reported to improve in vitro embryo development. DHA, the longest member of this family, can be produced from ALA and EPA; the efficiency of. Among their physiological roles, n-3 PUFA affect reproduction in cattle [7, 8]. An n-3 PUFA diet directly affected the cleavage rate and tended to increase the blastocyst rate after in vitro maturation (IVM) and in vitro fertilization (IVF) in cows [13, 14].
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