title: Canine sperm vitrification with sucrose: effect on sperm function.
abstract: The ability of sucrose to protect spermatozoa against mitochondrial damage, artificial acrosome reaction and DNA fragmentation during ultra-rapid cryopreservation in canine sperm was investigated. Swim-up selected spermatozoa of second-fraction semen were vitrified with different concentrations of sucrose (0.1, 0.25 and 0.4 m) in proportion 1 : 1 v/v with HTF-BSA 1%. From each group, 30-μl suspensions of cells were dropped directly into liquid nitrogen and stored for at least 24 h. Cells were thawed by submerging the spheres in HTF with 1% BSA at 37 °C. The number of progressively motile spermatozoa was significantly higher in the sucrose 0.25 m + HTF-BSA 1% (42.5 ± 2.3%, P < 0.01) than in HTF only (1.66 ± 0.3%). The same combination of sucrose 0.25 m + HTF-BSA 1% (42.7 ± 1.5%) had a stronger cryoprotective effect on the integrity of mitochondrial membrane potential (P < 0.05) and decreased the DNA fragmentation (2.8 ± 0.5%) as compared with HTF only (1.93 ± 0.6% and 5.6 ± 0.6% respectively). With respect to acrosome-reacted spermatozoa, no significant difference was found between the groups investigated (P > 0.05). It is concluded that sucrose, a nonpermeable cryoprotectant, can effectively preserve important physiological parameters such as mitochondrial membrane potential and DNA integrity during ultra-rapid cryopreservation.
authors.affiliation: BIOREN-CEBIOR, Universidad de La Frontera, Temuco, Chile.  rsanchez@ufro.cl
authors.forename: R
authors.initials: R
authors.lastname: Sánchez
authors.forename: J
authors.initials: J
authors.lastname: Risopatrón
authors.forename: M
authors.initials: M
authors.lastname: Schulz
authors.forename: J
authors.initials: J
authors.lastname: Villegas
authors.forename: V
authors.initials: V
authors.lastname: Isachenko
authors.forename: R
authors.initials: R
authors.lastname: Kreinberg
authors.forename: E
authors.initials: E
authors.lastname: Isachenko
mesh_terms: D000818:Animals; D015925:Cryopreservation; D003451:Cryoprotective Agents; D053938:DNA Fragmentation / Q000187:drug effects; D004285:Dogs; D008297:Male; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D012662:Semen Preservation / Q000379:methods; D013094:Spermatozoa / Q000187:drug effects / Q000502:physiology*; D013395:Sucrose / Q000494:pharmacology*; D058989:Vitrification
chemical_list: D003451:Cryoprotective Agents; D013395:Sucrose