Exogenous cholesterol prevents cryocapacitation like changes, membrane fluidity and enhances in-vitro fertility in bubaline spermatozoa.

A study was conducted to determine the optimum dosage of the exogenous cholesterol-loaded-cyclodextrins (CLC) to get maximum cryoprotection for bubaline spermatozoa. In the present study, 120×106 spermatozoa were incubated in 2, 3 and 4 mg of CLC as grouped as Gr II, III and IV, respectively and sperm progressive motility, intracellular Ca2+ , capacitation status by protein tyrosine phosphorylation (PTP) assay and zona binding percent (ZBP) and cleavage rate (CR) of the cryopreserved buffalo spermatozoa by in-vitro fertility assay were assessed in comparison with an untreated control group (Gr I). Results revealed that there was a significant (P<0.05) linear decrease of percentage of sperm population with higher intracellular Ca2+ and percentage of sperm population with medium or high capacitated by PTP in CLC treated from 2 to 3 mg and then increased to 4 mg/120×106 spermatozoa whereas sperm progressive motility, percentage of sperm population with low capacitated , ZBP and CR were increased significantly (P<0.05) in sperm population treated from 2 to 3 mg CLC and then decreased to 4 mg/120×106 spermatozoa. The study has clearly indicated that CLC at 3 mg/120×106 spermatozoa has maximum beneficial effects in protection of sperm progressive motility, membrane fluidity (low intracellular Ca2+ ); prevention of cryocapacitation (low capacitation pattern in immunolocalization) and enhancement of in-vitro ZBP and CR. Post thaw motility of the CLC treated sperm has shown positively significant (p<0.05) correlation with sperm population with low intracellular Ca2+ , low capacitated sperm population , ZBP and CR, whereas it was negatively (p<0.05) correlated with sperm population with high intracellular Ca2+ , medium or high capacitated sperm . The present study has revealed for the first time that incubation of spermatozoa with CLC of higher dose (>3 mg/120x106 spermatozoa) had adverse effects on sperm cryopreservation, although incubation of sperm with 3 mg/120 million prior to processing had minimised the freezing-thawing associated damages in bubaline species.

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