Seasonal differences in ram seminal plasma revealed by partition in an aqueous two-phase system |
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| Affiliation: | 1. Institute of Organic Chemistry with Centre of Phytochemistry, Bulgarian Academy of Sciences, 1113 Sofia, Bulgaria;2. Faculty of Pharmacy, Medical University of Sofia, 2 Dunav Str., 1000 Sofia, Bulgaria;3. Research Centre for Natural Sciences, Institute of Materials and Environmental Chemistry, Hungarian Academy of Sciences, 1117 Budapest, Magyar tudósok körútja 2, Hungary;1. Center for Research and Development Bioresources, Organization for Research Promotion, Osaka Prefecture University, Sakai, Osaka 599-8570, Japan;2. Department of Applied Life Sciences, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Sakai, Osaka 599-8531, Japan;3. Institute of Food Sciences, Nakagaki Consulting Engineer and Co., Ltd, Nishi-ku, Sakai, Osaka 593-8328, Japan;4. Faculty of Human Development, Department of Food and Nutrition Management Studies, Soai University, Osaka 559-0033, Japan;5. Department of Nutrition, Osaka Prefecture University, Habikino, Osaka 583-8555, Japan;6. Department of Chemistry, Biology and Marine Science, Faculty of Science, University of the Ryukyus, Nishihara, Okinawa 903-0213, Japan;7. Iga Research Institute, Iga Regional Satellite Campus, Mie University, Iga, Mie 518-0131, Japan;1. Chimie Analytique des Molécules Bioactives et Pharmacognosie, Institut Pluridisciplinaire Hubert Curien (UMR 7178 CNRS/UDS), 74 route du Rhin, 67400, Illkirch, France;2. Laboratoire A. Vogel, 5A Rue Lavoisier, 68000, Colmar, France;1. Faculty of Engineering and Science, University of Greenwich, Central Avenue, Chatham Maritime, Chatham, Kent ME4 4TB, UK;2. Faculty of Pharmaceutics Department, H.K. College of Pharmacy, Relief Road, Oshiwara, Jogeshwari West, Mumbai 400102, Maharashtra, India;3. Department of Pharmacy (Chemistry), School of Life Sciences, University of Sussex, Falmer, Brighton BN1 9QJ, UK;4. Drug Applied Research Center, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran;1. School of Science, College of Science, Engineering and Health, RMIT University, Melbourne, VIC 3001, Australia;2. Nanobiotechnology Laboratory, RMIT University, Melbourne, VIC 3001, Australia |
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| Abstract: | Seminal plasma plays an important role in maturation of spermatozoa through hormonal, enzymatic and surface-modifying events. We have previously shown that adsorption of seminal plasma proteins (SPPs) to the sperm cell surface partially restores the functional characteristics of damaged spermatozoa, reproducing those of live cells. In the present report, we investigate the hypothesis that seasonal differences in seminal plasma could affect its ability to recover membrane integrity of cold-shocked sperm. The effect of seminal plasma proteins, obtained in breeding (bsSPPs) and non-breeding (nbsSPPs) season, on cold-shocked ram spermatozoa previously freed from seminal plasma, was analysed by centrifugal counter-current distribution (CCCD) in an aqueous two-phase system as well as membrane integrity determination by fluorescence markers. Cold-shock treatment greatly lowered cell viability in both breeding and non-breeding season spermatozoa. The cold-shocked sperm viability obtained was approximately 20%. The loss of heterogeneity and the decrease in viability revealed by CCCD analysis was reversed by the addition of increasing amounts of bsSPP, which induced restoration of the surface characteristics of viable-like spermatozoa, as well as an increase in the number of recovered viable sperm. However, this restoring effect was much lower when nbsSPPs were added, even in a sixfold higher concentration than used with bsSPPs. Incubation of cold-shocked cells with both kinds of proteins performed in both seasonal periods, showed that the recovering effect was related to the season when the plasma sample was obtained rather than to the semen season. The addition of bsSPPs to cold-shocked sperm accounted for a nearly 50% reversion for both studied breeding seasons. However, the reversion percentages obtained with nbsSPPs were significantly lower (P<0.05) than those found with bsSPPs in both studied seasonal periods. This different reversion capacity of bsSPPs and nbsSPPs was related to a different protein composition, as revealed by comparative sodium dodecyl sulfate–polyacrylamide gel electrophoresis analysis. The bands of 20, 21, 24, 36 and 67 kDa of the bsSP sample profile decreased in winter–spring SP, and were even less intensely stained in summer SP. Densitometric analysis of the stained gel patterns allows automatic comparison among the separated bands, and revealed an important decrease in the content of several bands. The 21.5 kDa band showed the highest decrease, lowering to 14% in June–August plasma with respect to the value obtained in September–December plasma. |
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