Cryopreservation of cynomolgus macaque (Macaca fascicularis) sperm with glycerol and ethylene glycol, and its effect on sperm-specific ion channels - CatSper and Hv1

- Our results indicated that the cryoprotection of EG for sperm motility, acrosomal integrity, DNA integrity and mitochondrial membrane potential was equal to Gly, which confirmed that EG could be used as an alternative CPA for cynomolgus sperm freezing.
- Our studies revealed for the first time that cryopreservation either by Gly or EG decreased the quantity of sperm membrane ion channels CatSper 1 and Hv1.
- The study will be beneficial to understand the biological process involved in sperm cryopreservation of nonhuman primates and contribute to improve cryopreservation protocols than can maintain sperm function and fertilizing ability.

The cryoprotective agent (CPA) is one of the most important factors that affects the cryosurvival of sperm. The aim of the present study was to compare two different CPAs, glycerol (Gly) and ethylene glycol (EG), on the cryopreservation of cynomolgus macaques sperm and evaluate the effects of cryopreservation on sperm motility, acrosomal integrity, DNA integrity, mitochondrial function and the sperm membrane ion channels CatSper and Hv1. Compared to fresh sperm, cryopreservation with either 0.7 M Gly or EG decreased the sperm motility (79.8 ± 1.5% Vs. 47.3 ± 1.8% and 47.6 ± 1.4%), acrosomal integrity (89.6 ± 1.2% Vs. 80.1 ± 1.8% and 79.6 ± 1.7%), DNA integrity (91.9 ± 0.7% Vs. 82.9 ± 1.0% and 82.3 ± 1.0%) and mitochondrial membrane potential (87.9 ± 1.8% Vs. 70.6 ± 2.7% and 67.9 ± 2.5%) and the quantity of the CatSper and Hv1 channels determined by Western Blot (p < 0.05), and EG showed equal cryoprotection to cynomolgus sperm in all of the sperm parameters. Our results indicated, for the first time, that cryopreservation decreases the quantity of sperm membrane ion channels (CatSper and Hv1), which might be one of the reasons that frozen sperm have a low fertilizing ability. The study will be beneficial to understand the biological process involved in sperm cryopreservation of nonhuman primates and contribute to improving cryopreservation protocols than can maintain sperm function and fertilizing ability.