title: Improvement of European eel sperm cryopreservation method by preventing spermatozoa movement activation caused by cryoprotectants.
abstract: Sperm production has been obtained from European and Japanese eels, but its quality and quantity tend to be changeable. So, its cryopreservation has been tried in both species. Dimethyl sulfoxide (Me(2)SO) is the best cryoprotectant for European eel sperm, but increases the medium osmolality, inducing the activation of spermatozoa motility. To avoid this, different combinations of pH (6.5 and 8.5) and NaHCO(3) concentrations (20, 40 and 80mM) were tested with two Me(2)SO concentrations (5% and 10%). Foetal bovine serum (FBS, 25%v/v) was added as a membrane protector to all the freezing media used in the different experiments. The highest Me(2)SO and NaHCO(3) concentrations at pH 6.5 caused the best post-thawing motility (26+/-4%). A second experiment was carried out testing media with Me(2)SO 10% with additional NaHCO(3) concentrations (100 and 120 mM). The highest post-thawing motility (38+/-3%) was found in the media containing NaHCO(3) 100mM, but no significant difference was observed compared with the best in the previous experiment (NaHCO(3) 80 mM). In a parallel experiment, aiming to improve the protection against the cryopreservation process, bovine serum albumin (BSA, 5%w/v) was added instead of FBS. Lower motilities were registered with BSA as membrane protector. Spermatozoa activation caused by addition of Me(2)SO can be prevented using high NaHCO(3) concentrations, improving the cryopreservation process. This effect seems be based on some of the products dissociated from NaHCO(3) in aqueous solution, affecting the intracellular pH, essential in the sperm motility.
authors.affiliation: Instituto de Ciencia y Tecnología Animal, Universidad Politécnica de Valencia, Valencia, Spain.
authors.forename: D S
authors.initials: DS
authors.lastname: Peñaranda
authors.forename: L
authors.initials: L
authors.lastname: Pérez
authors.forename: V
authors.initials: V
authors.lastname: Gallego
authors.forename: M
authors.initials: M
authors.lastname: Jover
authors.forename: J F
authors.initials: JF
authors.lastname: Asturiano
mesh_terms: D000811:Anguilla / Q000502:physiology*; D000818:Animals; D017756:Aquaculture / Q000379:methods; D002470:Cell Survival / Q000187:drug effects; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary*; D003451:Cryoprotective Agents / Q000009:adverse effects*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D008297:Male; D012662:Semen Preservation / Q000379:methods / Q000662:veterinary*; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D017693:Sodium Bicarbonate / Q000494:pharmacology; D013081:Sperm Motility / Q000187:drug effects*; D013094:Spermatozoa / Q000187:drug effects* / Q000502:physiology
chemical_list: D003451:Cryoprotective Agents; D012710:Serum Albumin, Bovine; D017693:Sodium Bicarbonate; D004121:Dimethyl Sulfoxide