title: Osmotic tolerance of avian spermatozoa: influence of time, temperature, cryoprotectant and membrane ion pump function on sperm viability.
abstract: Potential factors influencing sperm survival under hypertonic conditions were evaluated in the Sandhill crane (Grus canadensis) and turkey (Meleagridis gallopavo). Sperm osmotolerance (300-3000 mOsm/kg) was evaluated after: (1) equilibration times of 2, 10, 45 and 60 min at 4 degrees C versus 21 degrees C; (2) pre-equilibrating with dimethylacetamide (DMA) or dimethylsulfoxide (Me2SO) at either 4 degrees C or 21 degrees C; and (3) inhibition of the Na+/K+ and the Na+/H+ antiporter membrane ionic pumps. Sperm viability was assessed using the eosin-nigrosin live/dead stain. Species-specific differences occurred in response to hypertonic conditions with crane sperm remaining viable under extreme hypertonicity (3000 mOsm/kg), whereas turkey sperm viability was compromised with only slightly hypertonic (500 mOsm/kg) conditions. The timing of spermolysis under hypertonic conditions was also species-specific, with a shorter interval for turkey (2 min) than crane (10 min) sperm. Turkey sperm osmotolerance was slightly improved by lowering the incubation temperature from 21 to 4 degrees C. Pre-equilibrating sperm with DMA reduced the incidence of hypertonic spermolysis only in the crane, at both room and refrigeration temperature. Inhibiting the Na+/K+ and the Na+/H+ antiporter membrane ion pumps did not impair resistance of crane and turkey spermatozoa to hypertonic stress; pump inhibition actually increased turkey sperm survival compared to control sperm. Results demonstrate marked species specificity in osmotolerance between crane and turkey sperm, as well as in the way temperature and time of exposure affect sperm survival under hypertonic conditions. Differences are independent of the role of osmotic pumps in these species.
authors.affiliation: USDA, ARS, ANRI, Biotechnology and Germplasm Laboratory, Beltsville, MD 20705, USA.
authors.forename: Juan M
authors.initials: JM
authors.lastname: Blanco
authors.forename: Julie A
authors.initials: JA
authors.lastname: Long
authors.forename: George
authors.initials: G
authors.lastname: Gee
authors.forename: Ann M
authors.initials: AM
authors.lastname: Donoghue
authors.forename: David E
authors.initials: DE
authors.lastname: Wildt
mesh_terms: D000081:Acetamides / Q000494:pharmacology; D000584:Amiloride / Q000494:pharmacology; D000818:Animals; D001717:Birds; D002470:Cell Survival; D015925:Cryopreservation / Q000662:veterinary*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D006982:Hypertonic Solutions / Q000494:pharmacology*; D016623:Ion Pumps / Q000502:physiology; D008297:Male; D009994:Osmolar Concentration; D010042:Ouabain / Q000494:pharmacology; D017923:Sodium-Hydrogen Exchangers / Q000187:drug effects / Q000502:physiology; D000254:Sodium-Potassium-Exchanging ATPase / Q000037:antagonists & inhibitors; D013045:Species Specificity; D013094:Spermatozoa / Q000187:drug effects / Q000502:physiology*; D013696:Temperature; D013997:Time Factors; D014422:Turkeys
chemical_list: D000081:Acetamides; D003451:Cryoprotective Agents; D006982:Hypertonic Solutions; D016623:Ion Pumps; D017923:Sodium-Hydrogen Exchangers; D010042:Ouabain; D000584:Amiloride; D000254:Sodium-Potassium-Exchanging ATPase; C013959:dimethylacetamide; D004121:Dimethyl Sulfoxide