The Biochemical Characteristics of Boar Seminal Plasma during High Ejaculation Frequency (original) (raw)
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Molecular Human Reproduction, 1996
The internal pH (pHi) of human spermatozoa was measured by the fluorescent indicator 2,7-bicarboxyethyl-5,6carboxyfluorescein acetoxymethyl ester (BCECF-AM) and the distribution of the radioactive [ 14 C]-methylamine under different external ionic conditions. The effect of the addition of progesterone and human follicular fluid (HFF) on the spermatozoa pHi was also analysed. The pHi values obtained were almost identical with the two probes used. In sodium (NaM) and potassium (KM) media, a linear relationship between the internal and external pH was observed. In NaM, the pHi values were -0.4 pH unit less than the external pH. In KM, the pHi measured was higher than in NaM and only slightly inferior to the external pH (0.1-0.2 pH unit). Addition of 10 fiM progesterone, oestradiol 170 or 20% HFF to spermatozoa incubated at pH 7.2 in NaM did not induce any rapid variation of the BCECF fluorescence or change in the accumulation of methylamine. A slight change in pH (-0.5 units) occurred with progesterone after 15 min. As a control, addition of 10 mM of NH 4 CI induced a rapid alkalinization (0.4 pH unit) of the cell interior while 10 mM lactate produced only a slight acidification (-0.2 pH unit). Under the same conditions (NaM, pH 7.2), the pHi of the spermatozoa prepared by Percoll gradient was found more acidic by 0.2 pH unit than washed unfractionated spermatozoa. Progesterone, oestradiol 170 and HFF had no effect on the pHi of these spermatozoa. The results obtained in this study show that it is possible to measure accurately the internal pH of human spermatozoa. Internal pH was found to be dependent upon the pH of the external medium and a quasi-linear relationship exists between the internal and external pH, suggesting no specific pH regulatory mechanisms. Our data suggest instead that the protons, under our experimental conditions, are passively distributed. Progesterone, oestradiol 170 and HFF, known to promote both capacitation and the acrosome reaction, do not act through a rapid pHi change.
Reproduction in Domestic Animals, 1995
Contents This study was performed to monitor biochemical changes in boar spermatozoa following two systems of semen-collection frequency. A total of eight monthly series of semen collections were used. Microscopic analyses of semen quality were accompanied by the following measurements of the activity of selected enzymes in order to assess integrity of spermatozoan structures: total acrosin activity for the acrosome, AspAT activity in spermatozoa for the middle piece, lipid peroxidation (LPO) and the H-33258 (H0258) for the plasmolemma. Further radioisotope measurements of 3H-actinomycine (3H-AMD) incorporation into the chromatin were used as an index of chromatin stability. The results of this study showed that long-term high ejaculation frequency leads to a gradual deterioration of the biological value of the spermatozoa and induces changes in the essential indices of semen quality (p < 0.01). Die Anderungen der Enzymaktivitat wurden von Integritatsanderungen der Zellmembran begleitet, was sich in einer (p s 0.01). intensiven HO-258-Bindung zeigte. Im Zellkern wurde eine sukzessive Hyperstabilisierung (die Senkung der 3H-AMD-Bindung) des Chromatins festgestelt. Die besprochenen Veranderungen waren individuell unterschiedlich. Es wird gefolgert, daR durch die hohen Samenentnahmefrequenzen die Kaskade der spezifischen biochemischen Spermienveranderungen, die der Apoptose der somatischen Zellen ahnlich sind, stimuliert werden.
Journal of Veterinary Medicine, 2013
Mutual relationships between selected chemical elements (Na, K, Fe, Cu, Mg, and Zn), basic motility characteristics (motility and progressive motility), and markers of the oxidative balance (superoxide dismutase, catalase, glutathione, albumin, and malondialdehyde) were investigated in bovine seminal plasma and spermatozoa. Computer assisted sperm analysis was used to assess the motility parameters; mineral concentrations were determined by the voltammetric method and flame absorption spectrophotometry; antioxidants and malondialdehyde were evaluated by UV/VIS spectrophotometry. Concentrations of chemical elements in both seminal fractions were in the following descending order: Na > K > Zn > Mg > Fe > Cu. Higher amounts of all minerals and nonenzymatic antioxidants were detected in the seminal plasma (P<0.01; P<0.001), while higher MDA concentration and activity of enzymatic antioxidants were recorded in the cell lysates (P<0.01; P<0.001). Na, Fe, Cu, Mg,...
Andrologia, 2009
Acid phosphatase was measured in the semen of normozoospermic, oligozoospermic, azoospermic and asthenoteratozoospermic men: there was no significant difference between oligozoospermic and control groups whereas we observed an increase in enzyme levels in the azoospermic group and a reduction in the asthenoteratozoospermic subjects. Furthermore, an isoelectric focusing method for the separation of isoenzymes was developed. Only prostatic isoenzymes 2 and 4 were found in semen. Isoenzyme 2 sialylation profile was determined in the four groups. The sperm count did not influence this profile. These findings are discussed. Auswertung der sauren Phosphatase-lsoenzyme im Spermaplasma bei Normozoospermie, Oligozoospermie, Azoospermie u nd Asthenozoosperm ie Zusammenfassung: Bei Mannern mit Normozoospermie, Oligozoospermie, Azoospermie und Asthenozoospermie wurde die saure Phosphatase im Sperma bestimmt: Es ergab sich kein signifianter Unterschied zwischen Oligozoospermie und den Kontrollgruppen, wahrend ein Anstieg der Enzymwerte in der Azoospermie-Gruppe und eine Reduzierung derselben bei Asthenozoospermie beobachtet werden konnte. Dariiber hinaus wurde eine isoelektrische Fokussierungs-Methode fur die Trennung der Isoenzyme entwickelt. Lediglich die Prostata-Isoenzyme 2 und 4 wurden im Samen gefunden. In den vier Gruppen wurde das Isoenzym-2-Sialylations-Profii bestimmt. Die Spermatozoendichte beeinfluate dieses Profil nicht.
Impact of Seminal Chemical Elements on the Oxidative
2013
Mutual relationships between selected chemical elements (Na, K, Fe, Cu, Mg, and Zn), basic motility characteristics (motility and progressive motility), and markers of the oxidative balance (superoxide dismutase, catalase, glutathione, albumin, and malondialdehyde) were investigated in bovine seminal plasma and spermatozoa. Computer assisted sperm analysis was used to assess the motility parameters; mineral concentrations were determined by the voltammetric method and flame absorption spectrophotometry; antioxidants and malondialdehyde were evaluated by UV/VIS spectrophotometry. Concentrations of chemical elements in both seminal fractions were in the following descending order: Na > K > Zn > Mg > Fe > Cu. Higher amounts of all minerals and nonenzymatic antioxidants were detected in the seminal plasma (< 0.01; < 0.001), while higher MDA concentration and activity of enzymatic antioxidants were recorded in the cell lysates (< 0.01; < 0.001). Na, Fe, Cu, Mg, and Zn were positively correlated with the motility and antioxidant parameters (< 0.05; < 0.01; < 0.001). Inversely, K exhibited the positive associations with malondialdehyde (< 0.05). This study demonstrates that most chemical elements are integral components of bovine semen and are needed for the protection against oxidative stress development.
Modification of human sperm metabolism by the induced release of intracellular zinc
Life Sciences, 1975
Incubation of washed human spermatozoa in the presence of 6 mM histidine induces a release of about 758 of the zinc bound to the cells. Zinc release induced by the presence of histidine was accompanied by : 1) a significant increase in the utilization of exogenous 14 C-labelled glucose, which is reflected in an increase in the production of 1 02 , 2) s small but significant decrease (-148) in the utilization of fructose when this sugar is added as exogenous substrate, and 3) a highly significant decrease in the endogenous sperm phospholipids (>308), an effect which is not inhibited by the addition of exogenous substrates . This behavior of human spermatozoa resembles that previously described for invertebrate spermatozoa and seems to be related to the regulation of energy metabolism and probably to sperm capacitation .
Human sperm hyperactivation and capacitation as parts of an oxidative process☆
Free Radical Biology and Medicine, 1993
Capacitation of spermatozoa is essential for fertilization and is visually characterized by hyperactivated motility. Previous reports have shown that foetal cord serum (FCS) and superoxide anion, 02, can trigger human sperm hyperactivation (HA) and capacitation and that superoxide dismutase (SOD) could prevent these processes. We investigated further the role of 02-and FCS components in human sperm HA and capacitation. Percoll-washed spermatozoa were incubated, at 37°C, in Ham's F-l0 medium with 7.5% of FCS, dialyzed FCS (> 12 kD), ultrafiltrate from FCS (FCSu; < 3 kD), or xanthine + xanthine oxidase + catalase (X + XO + cat). Spermatozoa incubated with FCSu were also supplemented with catalase to prevent the loss of motility often observed after 2-3 h of incubation. FCS and dialyzed FCS induced significant levels of HA (10 +_ 1% and 7.7 _+ 0.7%, respectively) that were, however, lower than those observed with FCSu (19 +_ 1%) or X + XO + cat (16 _+ 2%). Similar results were obtained when the lysophosphatidylcholine-induced acrosome reaction (LPC-AR, a measure of sperm capacitation) was evaluated. The presence of SOD in the incubation medium blocked the induction of HA and capacitation by FCS, FCSu, X + XO + cat, as well as the spontaneous HA and capacitation. The enzymatic activity of SOD was needed for the prevention of these processes. Desferrioxamine, up to 100 t~M, had no effect on HA and LPC-AR induced by FCSu and X + XO + cat. Addition of SOD to already hyperactivated spermatozoa reversed the HA. These data suggest that spermatozoa need a sustained O~-generation to maintain HA and proceed to capacitation. We hypothesize that FCSu or the O~-generated by X + XO + cat activate enzymes, possibly a reduced nicotinamide adenine dinucleotide phosphate [NAD(P)H] oxidase at the level of sperm membrane.
Relation between seminal quality and oxidative balance in sperm cells
Acta Urológica Portuguesa, 2016
Objectives: Infertility is a clinical disorder affecting approximately 15% of reproductive-aged couples worldwide. Recently, the influence of oxidative stress (OS) in decreased semen quality has been discussed. OS corresponds to an imbalance between oxidants and antioxidants defenses, present in the organism. High levels of reactive oxygen species (ROS) damage biomolecules present in sperm cells and may lead to the loss of membrane integrity, DNA fragmentation or even to death by apoptosis. This study aimed to evaluate the correlation between human semen clinic parameters and parameters that assessed the presence of OS. Material and methods: A total of 32 semen samples, obtained from a randomized group of donors, were included in this study. Basic semen parameters were analyzed according to the WHO's guidelines. The total antioxidant capacity of sperm cells was measured as well as the expression of certain antioxidant proteins, namely superoxide dismutase (SOD) and glutathione peroxidase 4 (GPx4), by colorimetric techniques and immunoblotting, respectively. The effect of ROS in spermatozoa protein oxidation was analyzed by determining the presence of 3nitrotyrosine and carbonyl groups, by slot blot. Lipid peroxidation was evaluated, by performing the thiobarbituric acid reactive substances (TBARS) assay with colorimetric tests. Results: The results indicated that SOD was negatively correlated with viscosity (p = 0.035), volume (p = 0.004) and carbonyl groups presence (p = 0.005). This protein also showed a positive correlation with the presence of tail defects in sperm cells (p = 0.044). In turn, GPx4 showed