title: Post-thaw ATP supplementation enhances cryoprotective effect of iodixanol in rat spermatozoa.
abstract: BACKGROUND
  Successful cryopreservation of rat spermatozoa from various strains still remains a challenge. The objective of this study was to determine if combinations of OptiPrep™ (iodixanol) and adenosine 5'-triphosphate (ATP) can improve rat sperm function during the cryopreservation procedure.
  
  
  METHODS
  Epididymal rat spermatozoa were frozen under different OptiPrep™ concentrations (0, 1, 2, 3 or 4 %) and were diluted with media supplemented with or without 2 mM ATP after thawing. Post-thaw sperm motility, acrosomal membrane integrity (AMI) and mitochondrial membrane potential (MMP) were then evaluated. In addition, the effect of different OptiPrep™ concentrations on fresh and cooled rat spermatozoa was tested via motility.
  
  
  RESULTS
  There was no effect of OptiPrep™ on motility of fresh and cooled spermatozoa. The supplementation of 1 and 2 % OptiPrep™ increased motility of frozen spermatozoa at 10 min after thawing, while it did not improve motility of spermatozoa at 3 h after thawing in the absence of ATP. During incubation of thawed spermatozoa, the ATP addition protected time-dependent decrease in motility after thawing in OptiPrep™-treated samples. OptiPrep™ had no effect on AMI and MMP in frozen-thawed spermatozoa but combinations of OptiPrep™ and ATP improved MMP in frozen-thawed spermatozoa.
  
  
  CONCLUSIONS
  Iodixanol has cryoprotective effects during rat sperm freezing without any toxic effect. Moreover, the combinations of iodixanol and ATP have a beneficial role in maintaining function of frozen-thawed rat spermatozoa for long period of incubation post-thaw.
authors.affiliation: Department of Veterinary Pathobiology, College of Veterinary Medicine, University of Missouri-Columbia, 1600 East Rollins Street, Room W191, Columbia, MO, 65211, USA.
authors.forename: Suhee
authors.initials: S
authors.lastname: Kim
authors.affiliation: Department of Veterinary Pathobiology, College of Veterinary Medicine, University of Missouri-Columbia, 1600 East Rollins Street, Room W191, Columbia, MO, 65211, USA.
authors.forename: Sarah
authors.initials: S
authors.lastname: Hooper
authors.affiliation: Department of Veterinary Pathobiology, College of Veterinary Medicine, University of Missouri-Columbia, 1600 East Rollins Street, Room W191, Columbia, MO, 65211, USA.
authors.forename: Cansu
authors.initials: C
authors.lastname: Agca
authors.affiliation: Department of Veterinary Pathobiology, College of Veterinary Medicine, University of Missouri-Columbia, 1600 East Rollins Street, Room W191, Columbia, MO, 65211, USA. agcay@missouri.edu.
authors.forename: Yuksel
authors.initials: Y
authors.lastname: Agca
mesh_terms: D000177:Acrosome / Q000187:drug effects; D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000818:Animals; D015925:Cryopreservation / Q000379:methods; D003451:Cryoprotective Agents / Q000494:pharmacology; D008297:Male; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley; D055101:Semen Analysis; D012662:Semen Preservation / Q000379:methods*; D013081:Sperm Motility / Q000187:drug effects; D013094:Spermatozoa / Q000187:drug effects*; D014283:Triiodobenzoic Acids / Q000494:pharmacology*
chemical_list: D003451:Cryoprotective Agents; D014283:Triiodobenzoic Acids; D000255:Adenosine Triphosphate; C044834:iodixanol
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