Abstract
The effect of resveratrol on the in vitro maturation (IVM) of ovine (Ovis aries) oocytes and the development of handmade cloned embryos was evaluated. The nuclear maturation and reactive oxygen species (ROS) levels in the oocytes, as well as the early development and morphological cloned embryo quality, were evaluated under different resveratrol concentrations (0, 0.5, 2 and 5 μM). After IVM, no significant difference was observed in the maturation rate of oocytes treated with 0.5 μM (81.3 %) and 2 μM (72 %) resveratrol compared to that of the control group (0 μM) (74.2 %), but the rate significantly decreased at 5 μM (56 %) (p < 0.05). When the oocyte ROS levels were determined, no significant differences among the groups were observed (p > 0.05). For cloned embryo development, the embryos obtained from the oocytes treated with 0.5 μM resveratrol showed higher (p < 0.05) compacted morula rates (10.7 %) compared to the embryos obtained from the oocytes treated with 0, 2 and 5 μM (6.2, 0 and 0 %, respectively). Regarding embryo morphological quality, the embryos from the oocytes treated with 0.5 μM resveratrol showed a lower rate of poor quality morulae (4.7 %) in comparison to those treated with 0, 2 and 5 μM (23.8, 23.3 and 33.3 %, respectively) (p < 0.05). In conclusion, resveratrol showed no significant improvement on the IVM or ROS levels in domestic ovine oocytes. However, treatment with 0.5 μM resveratrol during IVM improved embryo quality and promoted morulae compaction of Ovis aries handmade cloned embryos.Figure 3. Different developmental stages of the HMC sheep embryos cultured in the WOW system. Cleaved embryos (a-d), 8‒16 blastomere embryos (e-h), morulae (i-l) and compact morulae (m-p) (200X).
Highlights
In vitro embryo production by somatic cell nuclear transfer (SCNT) is a reproductive biotechnology with great potential for the propagation of genetically valuable individuals, farm animal production and the preservation of endangered wild species.[1]
Our results revealed that resveratrol treatment during in vitro maturation (IVM) resulted in concentration-dependent effects
The 5 μM resveratrol treatment decreased the IVM rate significantly compared to other concentrations
Summary
In vitro embryo production by somatic cell nuclear transfer (SCNT) is a reproductive biotechnology with great potential for the propagation of genetically valuable individuals, farm animal production and the preservation of endangered wild species.[1]. A variant of the technique is handmade cloning (HMC), which does not use micromanipulators, making it an accessible and less complex technique.[9] Using this technique, porcine,[10] bovine[11] and ovine[12] clones have been obtained For both cloning techniques, efficiency is still low; depending on the species, only 1–5 % of the transferred embryos develop into viable offspring, and cloned animals have shown developmental problems in other cases.[13] These problems could be due to the complexity of the technique, which involves several factors that can affect embryo development.[14] In this regard, Camargo et al.[15] stated that oocyte in vitro maturation is the main factor for obtaining optimal in vitro embryo development
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