Abstract
BackgroundThe regulation of methyl-beta-cyclodextrin (MBCD) on cryodamage on X- and Y-sperm during cryopreservation of semen was investigated. The semen was collected from ten healthy bulls of proven fertility by an artificial vagina. The bovine sperm treated with MBCD fresh solution (0, 1, 5, 10, and 20 mM). The sperms were evaluated for viability and acrosome damage using flow cytometry. Moreover, X- and Y-sperm in frozen-thawed bovine semen were sorted by flow cytometry after Hoechst 33342-dyed, and the viability and acrosome damage of sperms were analyzed.ResultsSperm viability in frozen-thawed semen was decreased by MBCD (p < 0.05), also the acrosome damage of sperm was significantly increased (p < 0.05). Moreover, we sorted X- and Y-sperm from frozen-thawed bovine semen for observing the viability and acrosome damage on the separated X- and Y-sperm after MBCD treatment. Viability of X-sperm was significantly lower than that of Y-sperm (p < 0.05). Also, acrosome damage of X-sperm was significantly higher than Y-sperm (p < 0.05).ConclusionsMethyl-beta-cyclodextrin enhances damage of sperm in frozen-thawed bovine semen, and X-sperm is more sensitive than Y-sperm in cell damage. These results demonstrate that MBCD can inhibit viability of spermatozoa in frozen-thawed bovine semen (for X-sperm, especially).
Highlights
The regulation of methyl-beta-cyclodextrin (MBCD) on cryodamage on X- and Y-bearing sperm (Y-sperm) during cryo‐ preservation of semen was investigated
Sperm was damaged by MBCD in frozen‐thawed bovine semen Sperm viability in frozen-thawed bovine semen was decreased by MBCD (Fig. 1, p < 0.05)
Viability of the treated-MBCD samples were significantly decreased in frozen-thawed sperm (52.2 ± 1.8 %, 37.5 ± 2.1 %, 19.4 ± 1.1 %, 16 ± 0.5 %, and 7.7 ± 0.9, respectively)
Summary
The regulation of methyl-beta-cyclodextrin (MBCD) on cryodamage on X- and Y-sperm during cryo‐ preservation of semen was investigated. The sperms were evaluated for viability and acrosome damage using flow cytometry. X- and Y-sperm in frozen-thawed bovine semen were sorted by flow cytometry after Hoechst 33342-dyed, and the viability and acrosome damage of sperms were analyzed. In the process of spermatogenesis, X- and Y-sperm can be produced, and the sex of the embryo is determined by. The main function of sperm is to produce embryos through fertilizing the oocyte, and capacitation indicates the fertility of sperm. The difference in acrosome reaction between X- and Y-sperm during capacitation is important. Methyl-beta-cyclodextrin (MBCD), which is a cyclic heptasaccharide consisting of beta gluco-pyranose units effectively extracts sperm sterols, resulting in the disruption of detergent resistant sperm membranes [16, 17]
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