Resveratrol significantly improves the fertilisation capacity of bovine sex-sorted semen by inhibiting apoptosis and lipid peroxidation

Chong-Yang Li,Chang-Liang Yan,Jin-Ming Huang,Yun-Wei Pang,Wei-Hua Du,Hua-Bin Zhu,Xue-Ming Zhao,Pei-Pei Zhang,Hai-Sheng Hao,Hao-Yu Wang,Yan Liu,Ya-Han Zhao

doi:10.1038/s41598-018-25687-z

Abstract

The aim of this study was to test the effects of five different concentrations (0, 10−3, 10−4, 10−5, and 10−6 M) of resveratrol (Res) supplementation in bull sperm washing and fertilisation medium on levels of reactive oxygen species (ROS), phosphatidylserine (PS) externalisation, mitochondrial membrane potential (Δψm), ATP and malondialdehyde (MDA), acrosomal integrity, blastocyst rate, and blastocyst quality after in vitro fertilisation (IVF). The results for sex-sorted sperm from three bulls showed: (1) ROS and MDA levels in 10−3 M and 10−4 M Res groups were significantly lower than those of controls (P < 0.05); (2) the percentage of viable sperm, percentage of sperm with high Δψm, and the ATP content in 10−3 M and 10−4 M Res groups were significantly higher than those of controls (P < 0.05); (3) the percentage of viable sperm with acrosomal integrity, and the blastocyst percentage and quality of the 10−4 M Res group were significantly higher than those of controls (P < 0.05). In conclusion, 10−4 M Res supplementation in washing and fertilisation medium of sex-sorted bull sperm significantly decreased ROS, PS externalisation, and MDA, and protected mitochondrial function and acrosomal integrity, thereby increasing blastocyst percentage and quality following IVF.

Highlights

In the dairy cattle industry, the ability to control the sex of calves presents a significant advantage for maximising profit from cattle breeding and dairy farming
reactive oxygen species (ROS) is the main inducer of lipid peroxidation (LPO) in sperm because their plasma membranes are rich in polyunsaturated fatty acids, the principal target for oxidation[7], and this decreases sperm motility and viability[8]
The percentage of early necrotic sperm in the [10−3] M Res group (1.16 ± 0.13%, 8.80 ± 0.81%, and 6.12 ± 0.51%) and the [10−4] M Res group (4.97 ± 0.38%, 14.35 ± 1.42%, and 14.20 ± 1.13%) was significantly lower than that of the control group (19.83 ± 1.27%, 28.81 ± 2.74%, and 22.28 ± 2.14%; P < 0.05), but no significant differences were found between the percentage of early apoptotic sperm in the control groups (44.25 ± 4.16% to 51.49 ± 4.35%) and [10−4] M Res groups (43.31 ± 4.25% to 50.88 ± 4.52%; P > 0.05)

Summary

Introduction

In the dairy cattle industry, the ability to control the sex of calves presents a significant advantage for maximising profit from cattle breeding and dairy farming. More than 40% of embryo transfers across the world use embryos produced in vitro by in vitro fertilisation (IVF), and sex-sorted sperm plays an important role in the commercial production of sexed embryos by IVF3,4 Both flow cytometry sorting[5] and cryopreservation[6] greatly increase reactive oxygen species (ROS) levels in sperm. Resveratrol (Res) is a natural grape-derived phytoalexin[13] possessing stronger antioxidant activity than vitamins E and C, as well as lower toxicity[14] Res acts both in the initiation and propagation of the oxidative process[14], and can access peroxidised rigid membranes and increase membrane fluidity[12,15,16]. The results contribute to the development of an efficient method for improving the fertilisation capacity of sex-sorted bull sperm, and exploration of the mechanisms involved

Objectives

Methods

Results

Conclusion