Variation between and within boar ejaculates in terms of their ability to withstand freeze-thawing is a limitation for sperm cryopreservation. Consequently, searching for freezability markers not only in sperm but also in seminal plasma (SP) is imperative. The present study aimed to evaluate the relationship between cholesterol content, relative levels of NPC2 and AQN-1 at two different holding times (0 h: HT0 and 24 h: HT24) at 17 °C, and boar sperm freezability. Forty-five ejaculates were cryopreserved and subsequently classified as of good (GFE) or poor (PFE) freezability according to their post-thaw sperm viability and total motility. Prior to cryopreservation, relative abundances of two SP proteins (NPC2 and AQN-1) and cholesterol content in sperm and SP were determined through immunoblotting and colorimetric methods, respectively. These determinations were made after ejaculation (HT0) and after 24 h of storage at 17 °C (HT24). Two bands for NPC2 protein (16 kDa and 19 kDa) were identified. Relative amounts of the 16 kDa-band were significantly (P < 0.05) higher in poor (PFE) than in good (GFE) freezability ejaculates both at HT0 and HT24, whereas those of the 19 kDa-band were significantly (P < 0.05) higher in PFE than in GFE at HT24 only. In the case of AQN-1, no significant differences between GFE and PFE were observed. In addition, no variations in the cholesterol content of sperm and SP were observed either between HT0 and HT24 or between GFE and PFE. We can conclude that the content of two NPC2 isoforms in SP, but not of that of spermadhesin AQN-1, may be involved in the sperm resilience to withstand freeze-thawing procedures and may predict ejaculate freezability. While a possible mechanism through which NPC2 during HT could affect boar sperm cryotolerance is suggested to be related to its ability to bind the plasma membrane cholesterol, further research is warranted.
Keywords: AQN-1; Boar sperm; Cryopreservation; Freezabillity marker; NPC2.
Copyright © 2019 Elsevier Inc. All rights reserved.