ABSTRACT
The present study aimed to investigate the effect of different seminal plasma proteins (SPPs) on boar spermatozoa functional characteristics. We investigated the putative protective effect of SSPs on sperm cells motility and velocity, as well as on the integrity of the plasma membrane (PM) during low-temperature storage at 4 °C. SPPs fractions were obtained and purified by gel permeation chromatography (GPC). Nine fractions of SPPs were obtained and further characterized by 12% sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Sperm computer analysis (SCA) after incubation of spermatozoa with separated proteins revealed that fraction 6 consisting of low molecular weight (MW) proteins could preserve spermatozoa motility and velocity better when compared to those with higher MW. Two-dimensional (2D) elecrtophoretic analysis showed that fraction 6 contained proteins with the following MW and isoelectric point (pI): 16 kDa and pI 7.35, 18 kDa and pI 5.20, 19 kDa and pI 7.35, 26 kDa and pI 4.50, 26 kDa and pI 4.30, 29 kDa and pI 5.85.
ABSTRACT
Cigarette smoking is a serious health problem of our society. It is known that cigarette smoke is a cell mutagen and carcinogen, and that it may affect adversely male fertility. The possible detrimental effects on sperm cells are of great interest but the data available to support this statement are somewhat elusive. To approach this problem we examined conventional semen parameters, plasma membrane translocation of phosphatidylserine (PS) (annexin V/6-CFDA cell staining) and sperm DNA integrity (comet assay) in a group of healthy man smoking cigarettes on a regular basis. The results of the study were compared with the results of the same tests in healthy non-smoking donors. Significant difference in standard sperm parameters between the two groups was not found. Intensive expression of PS on the sperm plasma membrane surface (assayed by annexin V positive staining) was detected in the smokers group. There is a significant increase of population of apoptotic spermatozoa in ejaculates of smokers. Albeit DNA damages (high frequencies of double- and single- stranded DNA breaks) in spermatozoa of smokers are increased compared with non-smokers, but this difference is not statistically significant. Sperm DNA integrity of healthy smokers remains in the normal range, but a clear negative trend is observed, especially in respect of disturbance of plasma membrane phospholipid asymmetry.
