Ion-mediated resistance to osmotic changes of ram spermatozoa: the role of amiloride and ouabain.
The aim of this study was to evaluate whether the Na+/K+ and Na+/H+ exchange can maintain the function of fresh ram spermatozoa. We analyzed the quality parameters of spermatozoa from fresh ram ejaculates incubated in iso- (about 300 mOsm), hypo- (about 100 mOsm) and hyperosmotic (about 900 mOsm) media in the presence of ouabain a specific inhibitor of the Na+/K+ ATP-ase or amiloride, a specific inhibitor of the Na+/H+ antiporter. Ouabain increased the percentage of morphologically altered acrosomes in isoosmotic media (from about 10% to 15% in control to about 30% with 10(-4) M ouabain) and decreased the percentage of total motility (from about 80% in control to about 50% to 55% with 10(-4) M ouabain). Ouabain decreased the mean linearity component of motility and decreased the frequency of head displacement. The addition of ouabain increased the percentage of altered acrosomes in the hypo- and hyperosmotic media, although it did not modify viability in either media. Ouabain also increased the percentage of swollen tails in the hypoosmotic medium at higher concentrations of the inhibitor. Amiloride increased the percentage of altered acrosomes in all media although its effect was less intense than that of ouabain. In isoosmotic media, total motility was decreased from about 80% in control to about 65% with 10(-4) M amiloride. Motile spermatozoa incubated with amiloride showed a clear decrease of mean velocity and mean linearity and increased frequency of head displacement. In the hyperosmotic medium, adding amiloride decreased the percentage of viability and altered tails at concentrations as low as 10(-6) to 10(-5) M. Our results indicate that the active mechanisms which control Na+ transport play a significant role in the maintenance of function in ram spermatozoa subjected to different osmotic environments. These mechanisms may be important in maintaining ram sperm function both "in vivo" and "in vitro".
Peris
S
S
Unit of Reproduction, School of Veterinary Medicine, Autonomous University of Barcelona, Bellaterra, Spain.
Solanes
D
D
Peña
A
A
Enric-Rodríguez-Gil
J
J
Riga
T
T
D000177:Acrosome / Q000187:drug effects / Q000648:ultrastructure; D000584:Amiloride / Q000494:pharmacology*; D000818:Animals; D002412:Cations / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006982:Hypertonic Solutions; D007038:Hypotonic Solutions; D008297:Male; D009994:Osmolar Concentration; D010042:Ouabain / Q000494:pharmacology*; D012756:Sheep; D017923:Sodium-Hydrogen Exchangers / Q000037:antagonists & inhibitors; D000254:Sodium-Potassium-Exchanging ATPase / Q000037:antagonists & inhibitors; D012996:Solutions; D013081:Sperm Motility / Q000187:drug effects; D013082:Sperm Tail / Q000187:drug effects / Q000648:ultrastructure; D013094:Spermatozoa / Q000002:abnormalities / Q000502:physiology* / Q000648:ultrastructure
D002412:Cations; D004791:Enzyme Inhibitors; D006982:Hypertonic Solutions; D007038:Hypotonic Solutions; D017923:Sodium-Hydrogen Exchangers; D012996:Solutions; D010042:Ouabain; D000584:Amiloride; D000254:Sodium-Potassium-Exchanging ATPase