Relationship Between Toxicity of Cryoprotectants, Osmotic and Oxidative Stresses In Awassi Ram Sperm

Author:

Varışh Ömer1,Bozkaya Faruk2,Aydilek Nurettin3,Taşkın Abdullah4

Affiliation:

1. Department of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine, K??r??kkale University, K??r??kkale, Turkey

2. Department of Genetics, Faculty of Veterinary Medicine, Harran University, Şanl??urfa, Turkey

3. Department of Physiology, Faculty of Veterinary Medicine, Dicle University, Diyarbak??r, Turkey

4. Faculty of Health Sciences, Harran University, Şanl??urfa, Turkey

Abstract

BACKGROUND: The relationship between the toxicity of cryoprotectants and their osmotic and/or oxidative stresses remains to be further investigated . OBJECTIVE: To investigate the toxic effects of different cryoprotectants and osmotic stress on Awassi ram sperm and to determine the relationship between oxidative and antioxidative status of the sperm. MATERIALS AND METHODS: Pooled sperm samples were exposed to sucrose solutions of different concentrations (75 to 900 mOsm) and isosmotic condition (290-325 mOsm) was re-established by adding HEPES buffered Tyrode's lactate. Sperm samples were mixed with 0.5, 1.0 and 1.5 M of glycerol, methanol, 2-methoxyethanol, dimethylacetamide or 1,2propanediol for 5 min and returned to isosmotic condition. Sperm samples were exposed to cryoprotectants at 4°C for 2 hours and isosmotic conditions were re-established. Motility, viability, acrosome integrity and oxidative or antioxidative parameters were determined. RESULTS: Treatment with hypo- or hyperosmotic sucrose solution reduced motility and viability without affecting acrosome integrity. The addition and removal of glycerol and dimethylacetamide (1.0 or 1.5 M) decreased sperm motility, while cryoprotectants had no effect on viability except for 1.5 M glycerol. Chilling significantly reduced the motility and viability of the sperm, but not the acrosome integrity. Rapid addition or removal of cryoprotectants also did not affect the acrosome integrity. Cryoprotectants changed only the ceruloplasmin level, while there were significant post-chilling differences in lipid hydroperoxide, paraoxonase and ceruloplasmin levels. CONCLUSION: Cryoprotectants without other additives have limited protection and glycerol can be toxic to spermatozoa. The oxidative stress plays a role in cryoprotectant toxicity and chilling stress.

Publisher

CryoLetters Limited Liability Partnership

Subject

General Medicine

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