Effects Of Treating With OW Temperature On Spermatozoa Ubiquitin Levels And In Vitro Fertilization Of Pigs

The main purpose of this study was to explore the effects of treating with low te mperature on spermarozoa ubiquitin levels in vitro fertilization of pigs.CASA, SDS-PAGE and Western blot were performed, results are as follows:1. Sperm motility, acrosome membrane integrity and the integrity of the epididymal spermatozoa in the test group were all higher than that of the ejaculated spermatozoa.2. Curve velocity (VCL), average linear velocity (VSL) and average path velocity (VAP) of the epididymal spermatozoa were all higher than that of the ejaculated spermatozoa.3. SDS-PAGE results showed that spermatozoa protein species orquantity werechanged after cold shock and freezing-thawing treatment. While no differences were detected between ejaculated and epidydimal spermatozoa.4. Analyzed porcine sperm protein of different treatmentsby Western blot, results showed that ubiquitin existedat 30kDa、and 62kDa. Gray value analysis showed significant differences were found at 46kDa among various treatments. Ubiquitin at 62kDawas only found at fresh ejaculated and epidydimal spermatozoa. Ubiquitin was greatly reduced in cold shock treatment group than other treatments.5. Analyzed porcine sperm protein of freezing-thawing spermatozoa by Western blot. Results showed lowest gray value was observed in the -80℃ treated spermatozoa compared with other three treatments. -196℃ treated epidydimalspermatozoashowed highest gray value than other groups.6. Spermatozoa with four different treatments were used tofertilize mature procine oocytes, cleavage was observed after 2 days. Results showed cleavage rates were higher in the embryos fertilized with epidydimal spermatozoacompared with that with ejaculated spermatozoa; higher cleavage rateswere found in the cold shock epidydimal spermatozoa group when compared with that of cold shock epidydimal spermatozoa group.7. Spermatozoa with four different treatments were used tofertilize mature procine oocytes,cleavage was observed after 2 days. Results showed cleavage rates were higher in the embryos fertilized with epidydimal spermatozoatreated at -80℃ and-196℃ compared with that with treated ejaculated spermatozoa.