Resveratrol treatment during goat oocytes maturation enhances developmental competence of parthenogenetic and hand-made cloned blastocysts by modulating intracellular glutathione level and embryonic gene expression

Abstract

Purpose The aim of the present study was to determine whether supplementation of resveratrol, a stilbenoid antioxidant with therapeutic significance, influences goat (Capra hircus) oocyte maturation and subsequent embryonic development and expression of apoptosis and early embryonic development-related genes. Methods Five different concentrations of resveratrol (0.1, 0.25, 0.5, 2.0 and 5.0 μM) were used in in vitro maturation (IVM) medium. Cell tracker blue and 2′,7′-dichlorodihydrofluorescein diacetate (H 2 DCFDA) fluorescent stains were used to assay intracellular glutathione and reactive oxygen species levels in mature oocytes. Parthenogenetic activation and hand-made cloning were performed to check the developmental potential following resveratrol treatment. We used quantitative real-time PCR to analyze embryonic gene expression. Result Compared to control, no significant improvement was observed in nuclear maturation in resveratrol-treated groups and at 5.0 μM concentration maturation rate decreased significantly (P <0.05). But resveratrol treatment at the concentrations of 0.25, 0.5 μM significantly reduced intracellular ROS, and increased GSH concentrations. Oocytes treated with 0.25, 0.5 μM resveratrol when subsequently used for PA and HMC, higher extent of blastocyst yields were observed. Expression analysis of proapoptotic (Bax) gene in mature oocytes, cumulus cells, and HMC-derived blastocysts revealed lesser transcript abundances in various resveratrol-treated groups., however no change in the same was observed for antiapoptotic gene (Bcl2). Differential expression of genes associated with developmental competence and nuclear reprogramming was also observed in HMC-derived blastocysts. Conclusion Our results show that resveratrol treatment at optimum concentrations (0.25 and 0.5 μM) during IVM produced beneficial microenvironment within oocytes by increasing the intracellular GSH, decreasing ROS level and this in turn, stimulated embryonic development and regulated gene expression.

Oocyte maturation with royal jelly increases embryo development and reduces apoptosis in goats

“…The increased level of GSH was coupled with reducing the intracellular levels of ROS, oxidative stress-induced apoptosis, and altering the expression of oocytes' molecular markers during IVM (Kwak et al, 2012;Kim et al, 2015;Jin et al, 2016). In support to this assumption, addition of resveratrol during IVM of goat COCs has significantly reduced both intracellular ROS and expression of proapoptotic (Bax) gene while increasing GSH levels (Mukherjee et al, 2014).…”

Quantitative expression of antiapoptotic and proapoptotic genes in sheep ovarian follicles grown in vivo or cultured in vitro

“…Therefore, it is necessary to identify the factors/conditions that improve Bcl2 but reduce Bax expression in the cultured follicles in future studies. Mukherjee et al [35] reported that resveratrol reduces the expression of Bax genes in goat oocytes. Similarly, estradiol suppressed apoptosis in the rabbit CL by altering the expression of Bcl2 family of genes [36].…”

Protective effect of resveratrol against oxidative damage to ovarian reserve in female Sprague–Dawley rats

“…These effects of resveratrol seem more likely to be associated with increasing telomere length and telomerase activity and reduction of germ cell dysfunction. Mukherjee et al (2014) showed that the supplementation of resveratrol at optimum concentrations during IVM improves the microenvironment within oocytes by increasing the intracellular GSH and decreasing the ROS level, thus stimulating embryonic development, and regulating gene expression (Mukherjee et al, 2014). Several studies have evaluated the effect of resveratrol as a novel treatment modality for polycystic ovary syndrome.…”