Effect of trehalose- and sucrose-based extenders on equine sperm quality after vitrification: Preliminary results

C C Pérez-Marín; F D Requena; A Arando; S Ortiz-Villalón; F Requena; E I Agüera

doi:10.1016/j.cryobiol.2017.12.002

Effect of trehalose- and sucrose-based extenders on equine sperm quality after vitrification: Preliminary results

Cryobiology. 2018 Feb:80:62-69. doi: 10.1016/j.cryobiol.2017.12.002. Epub 2017 Dec 8.

Authors

C C Pérez-Marín¹, F D Requena², A Arando³, S Ortiz-Villalón⁴, F Requena², E I Agüera²

Affiliations

¹ Department of Animal Medicine and Surgery, University of Cordoba, Cordoba 14014, Spain. Electronic address: pv2pemac@uco.es.
² Department of Cell Biology, Physiology and Immunology, University of Cordoba, 14014 Cordoba, Spain.
³ Department of Genetics, University of Cordoba, Cordoba 14014, Spain.
⁴ Department of Animal Medicine and Surgery, University of Cordoba, Cordoba 14014, Spain.

PMID: 29229561
DOI: 10.1016/j.cryobiol.2017.12.002

Abstract

There has been a lack of research into equine sperm vitrification to date, but studies of other species suggest it may have significant potential. To evaluate the impact of various cryoprotectant agents (CPA) and vitrification on equine sperm quality, a controlled study was carried out. A total of 12 ejaculates were subjected to exposure to CPA and vitrification. Sperm was diluted in a range of CPA: fresh, control (BSA), sucrose (0.15M, 0.3M and 0.5M), trehalose (0.15M, 0.3M and 0.5M) and the combination of sucrose and trehalose (M1: 0.15M sucrose+0.5M trehalose; M2: 0.5M sucrose+0.15M trehalose). Sperm motility, viability, acrosome integrity and DNA fragmentation were assessed at the time of CPA exposure and after vitrification. The exposure of spermatozoa to various concentrations of sucrose and/or trehalose significantly reduced sperm motility, with lower concentrations resulting in higher sperm motility. Sperm viability and DNA fragmentation did not vary after exposure to CPA, but acrosome integrity fell significantly when spermatozoa were exposed to CPA with high osmolality. When spermatozoa were vitrified, motility values were significantly higher than those obtained during the exposure. Low concentrations of sucrose (0.15M and 0.3M) and trehalose (0.15M) showed the best progressive sperm motility. The vitrification-warmed procedure significantly reduced sperm viability and acrosome integrity, but DNA did not vary with any of CPA used. Equine sperm vitrification demonstrates a low capacity for preserving sperm motility, and extenders containing trehalose or sucrose at lower concentrations are associated with a better protective effect on sperm motility. After vitrification, acrosome and plasma membranes were severely impaired, while the DNA structure was maintained. Equine spermatozoa partially recover the motility after vitrification, but there is a need for further studies into the preservation of sperm membranes.

Keywords: Disaccharides; Spermatozoa; Stallion; Vitrified.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Acrosome / drug effects
Animals
Cell Membrane / drug effects
Cryopreservation / methods*
Cryopreservation / veterinary
Cryoprotective Agents / pharmacology*
DNA Fragmentation / drug effects
Horses
Male
Semen Preservation / methods*
Semen Preservation / veterinary
Sperm Motility / drug effects
Sucrose / pharmacology*
Trehalose / pharmacology*
Vitrification / drug effects*

Substances

Cryoprotective Agents
Sucrose
Trehalose