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Effect of linoleic acid supplementation on in vitro maturation, embryo development and apoptotic related gene expression in ovine.

ABSTRACT: BACKGROUND:Linoleic acid (LA) is a polyunsaturated fatty acid present in high concentrations in follicular fluid, when added to maturation culture media, it affects oocyte competence. OBJECTIVE:In the present study, we investigated effect of linoleic acid supplementation on in vitro maturation, embryo development and apoptotic related gene expression in ovine. MATERIALS AND METHODS:The experiments conducted on 450 ovine Cumulus-oocyte complexes (COCs) with homogenous ooplasm and more than two compact layers of cumulus cells. For in vitro maturation COCs were randomly allocated into four treatment groups for 24 hr period. Treatment groups were as follow: control maturation media, 0 µM LA, 50 µM LA, 100 µM LA and 200 µM LA. The cumulus cell expansion and blastocysts rates were recorded. Total RNA was isolated from embryo pools, reverse transcribed into cDNA, and subjected to apoptotic gene expression by real-time PCR. RESULTS:Highest concentration (200 µM/mL) of LA significantly decreased the rate of fully expanded cumulus cells 24 hr after in vitro maturation (IVM) and the percentage of blastocyste rate compared with the control (p<0.05). These inhibitory effects were associated with an increased in relative mRNA expression of Bax (Bcl-2- associated X) gene compared with controls. CONCLUSION:Data obtained in present study suggest that low concentration of LA used for maturation had no deleterious effect on subsequent embryonic development compared to high concentration of LA. Relative expression of Bcl-2 (B-cell lymphoma 2) and Bax in embryos seems to be associated with LA concentration.

SUBMITTER: Amini E

PROVIDER: S-EPMC4918772 | biostudies-literature | 2016 Apr

REPOSITORIES: biostudies-literature

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Effect of linoleic acid supplementation on in vitro maturation, embryo development and apoptotic related gene expression in ovine.

Amini Ebrahim E Asadpour Reza R Roshangar Leila L Jafari-Joozani Razi R

International journal of reproductive biomedicine 20160401 4

<h4>Background</h4>Linoleic acid (LA) is a polyunsaturated fatty acid present in high concentrations in follicular fluid, when added to maturation culture media, it affects oocyte competence.<h4>Objective</h4>In the present study, we investigated effect of linoleic acid supplementation on in vitro maturation, embryo development and apoptotic related gene expression in ovine.<h4>Materials and methods</h4>The experiments conducted on 450 ovine Cumulus-oocyte complexes (COCs) with homogenous ooplas ...[more]

PMID: 27351027

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Project description:BackgroundHeavy metal cadmium (Cd) is a widespread environmental contaminant with a potential toxicity that might negatively affect female reproduction and fertility. It has been reported that Cd exposure impaired the quality of oocytes and led to a defective maturation and fertilization, through oxidative stress induction. Resveratrol (Res) is a natural polyphenol with strong antioxidant properties that exhibited protective role in preventing oocyte redox homeostasis disruption and quality decline. Here, we explored whether the addition of Res to in vitro maturation (IVM) medium might act as a protection against Cd-induced toxicity on ovine oocyte maturation and fertilization. Firstly, we evaluated the effect of supplementing IVM medium with two different Res concentrations (1 and 2 μmol/L) on nuclear maturation and fertilization of oocytes matured under CdCl2 (2 μmol/L) exposure. Therefore, the concentration of 1 μmol/L Res was selected to analyse the effects of this compound on intracellular ROS levels, mitochondrial (mt) distribution and activity, chromatin configuration, cytoskeleton morphology, cortical granules (CGs) distribution and mRNA expression of genes associated with cellular response to oxidative stress (i.e. SIRT1, SOD 1, GPX1, GSR, CAT) in Cd-exposed in vitro matured oocytes.ResultsWe found that 1 μmol/L Res restored the reduced oocyte meiotic competence induced by Cd exposure as well as, Res sustained oocyte ability to be normally fertilized and decreased polyspermic fertilization at both tested concentrations. Moreover, we demonstrated that 1 μmol/L Res mitigated Cd-induced alterations of oocyte cytoplasmic maturation by reducing reactive oxygen species (ROS) accumulation, preventing mt dysfunction, maintaining the correct meiotic spindle and cortical F-actin assembly and the normal cortical granule distribution as well as up-regulating SIRT1, SOD1 and GPX1 genes.ConclusionsTaken together, our findings highlighted the beneficial influence exerted by Res in preventing Cd-induced disturbance of nuclear and cytoplasmic maturation and subsequent fertilization in ovine oocytes. Res treatment may help to establish defence strategies counteracting Cd-induced toxicity on the female gamete.

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Effect of linoleic acid supplementation on in vitro maturation, embryo development and apoptotic related gene expression in ovine.

Publications

Effect of linoleic acid supplementation on in vitro maturation, embryo development and apoptotic related gene expression in ovine.

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