scholarly journals Equine seminal plasma reduces sperm binding to polymorphonuclear neutrophils (PMNs) and improves the fertility of fresh semen inseminated into inflamed uteri

Reproduction ◽  
2004 ◽  
Vol 127 (5) ◽  
pp. 593-600 ◽  
Author(s):  
A S Alghamdi ◽  
D N Foster ◽  
M H T Troedsson
Keyword(s):  
Seminal Plasma ◽  
Skim Milk ◽  
Polymorphonuclear Neutrophils ◽  
Proteinase K ◽  
Polymorphonuclear Neutrophil ◽  
Sperm Binding ◽  
Uterine Inflammation ◽  
Semen Extender ◽  
Uterine Secretions

Seminal plasma (SP) is known to have immunosuppressive properties in several species. Equine SP has been reported to reduce or inhibit chemotaxis, phagocytosis and complement activity in vitro. The type and amount of the SP component that suppresses sperm–polymorphonuclear neutrophil (PMN) binding in vitro was determined, and the effect of such suppression on the fertility of mares inseminated in the presence of uterine inflammation, was analyzed. Sperm cells were suspended in either SP, semen extender or a mixture of both, and each was mixed with PMN-rich uterine secretions collected at 12 h after artificial insemination (AI). SP reduced binding between spermatozoa and PMNs significantly (P < 0.05). Fertile spermatozoa were suspended in SP or semen extender and used to inseminate mares 12 h after the induction of uterine inflammation. The pregnancy rate was normal (77%) when spermatozoa were suspended in SP, but was dramatically reduced to only 5% when spermatozoa were suspended in extender. The proteins from SP, blood plasma (BP) and a skim-milk-based semen extender (skim milk extender, SME) were precipitated by ammonium sulfate, resuspended in PBS and dialyzed. The effect of the precipitated proteins on sperm–PMN binding was compared with fresh, untreated SP. Both fresh SP, and isolated SP proteins reduced sperm–PMN binding (P < 0.001). Conversely, proteins isolated from either BP or SME did not reduce sperm–PMN binding. The different concentrations of SP proteins used showed a dose-dependent suppression of sperm–PMN binding. Concentrations of 1 mg/ml SP protein significantly reduced sperm–PMN binding and 6 mg/ml reduced the binding to a level similar to that observed with fresh whole SP (P < 0.001). Finally, SP protein digested with proteinase K resulted in the complete loss of SP suppressive activity confirming that the effective component is a proteinaceous substance.

Effect of donkey seminal plasma on sperm movement and sperm–polymorphonuclear neutrophils attachment in vitro

2013 ◽  
Vol 140 (3-4) ◽  
pp. 164-172 ◽  
Author(s):  
Jordi Miró ◽  
Karina Vilés ◽  
Wilber García ◽  
Jordi Jordana ◽  
Marc Yeste
Keyword(s):  
Seminal Plasma ◽  
Polymorphonuclear Neutrophils ◽  
Sperm Movement

Proteomic analysis of spermatozoa reveals caseins play a pivotal role in preventing short-term periods of subfertility in stallions

2022 ◽  
Author(s):  
Róisín Ann Griffin ◽  
Aleona Swegen ◽  
Mark A Baker ◽  
Rachel Ann Ogle ◽  
Nathan Smith ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Plasma Proteins ◽  
Low Fertility ◽  
High Fertility ◽  
Short Term ◽  
Sperm Binding ◽  
Merocyanine 540 ◽  
Seminal Plasma Proteins ◽  
Assessment Strategies

Abstract Stallions experience transient fluctuations in fertility throughout the breeding season. Considering pregnancy diagnoses cannot be ascertained until ~14 days post-breeding, the timely detection of decreases in stallion fertility would enhance industry economic and welfare outcomes. Therefore, this study aimed to identify the proteomic signatures reflective of short-term fertility fluctuations, and to determine the biological mechanisms governing such differences. Using LC–MS/MS, we compared the proteomic profile of semen samples collected from commercially “fertile” stallions, during high- and low-fertility periods. A total of 1702 proteins were identified, of which, 38 showed a significant change in abundance (p ≤ 0.05). Assessment of intra- and inter-stallion variability revealed that caseins (namely κ-, α-S1-, and α-S2-casein), were significantly more abundant during “high-fertility” periods, while several epididymal, and seminal plasma proteins (chiefly, epididymal sperm binding protein 1 [ELSPbP1], horse seminal plasma protein 1 [HSP-1] and clusterin), were significantly more abundant during “low-fertility” periods. We hypothesised that an increased abundance of caseins offers greater protection from potentially harmful seminal plasma proteins, thereby preserving cell functionality and fertility. In vitro exposure of spermatozoa to casein resulted in decreased levels of lipid scrambling (Merocyanine 540), higher abundance of sperm-bound caseins (α-S1-, α-S2-, and κ-casein), and lower abundance of sperm-bound HSP-1 (p ≤ 0.05). This study demonstrates key pathways governing short-term fertility fluctuations in the stallion, thereby providing a platform to develop robust, fertility assessment strategies into the future.

scholarly journals Characterization of an Antiviral Component in Human Seminal Plasma

2021 ◽  
Vol 12 ◽  
Author(s):  
Ran Chen ◽  
Wenjing Zhang ◽  
Maolei Gong ◽  
Fei Wang ◽  
Han Wu ◽  
...  
Keyword(s):  
Viral Infection ◽  
Cell Line ◽  
Seminal Plasma ◽  
Hela Cells ◽  
Sexual Transmission ◽  
Antiviral Effect ◽  
Rnase A ◽  
Proteinase K ◽  
Human Seminal Plasma

Numerous types of viruses have been found in human semen, which raises concerns about the sexual transmission of these viruses. The overall effect of semen on viral infection and transmission have yet to be fully investigated. In the present study, we aimed at the effect of seminal plasma (SP) on viral infection by focusing on the mumps viral (MuV) infection of HeLa cells. MuV efficiently infected HeLa cells in vitro. MuV infection was strongly inhibited by the pre-treatment of viruses with SP. SP inhibited MuV infection through the impairment of the virus’s attachment to cells. The antiviral activity of SP was resistant to the treatment of SP with boiling water, Proteinase K, RNase A, and DNase I, suggesting that the antiviral factor would not be proteins and nucleic acids. PNGase or PLA2 treatments did not abrogate the antiviral effect of SP against MuV. Further, we showed that the prostatic fluid (PF) showed similar inhibition as SP, whereas the epididymal fluid and seminal vesicle extract did not inhibit MuV infection. Both SP and PF also inhibited MuV infection of other cell types, including another human cervical carcinoma cell line C33a, mouse primary epididymal epithelial cells, and Sertoli cell line 15P1. Moreover, this inhibitory effect was not specific to MuV, as the herpes simplex virus 1, dengue virus 2, and adenovirus 5 infections were also inhibited by SP and PF. Our findings suggest that SP contains a prostate-derived pan-antiviral factor that may limit the sexual transmission of various viruses.

scholarly journals Inhibition of neutrophil chemotaxis by pig seminal plasma in vitro: a potential method for modulating post-breeding inflammation in sows

Reproduction ◽  
2001 ◽  
pp. 567-572 ◽  
Author(s):  
KJ Rozeboom ◽  
G Rocha-Chavez ◽  
MH Troedsson
Keyword(s):  
Blood Plasma ◽  
Seminal Plasma ◽  
Potential Method ◽  
Suppressive Effect ◽  
Sample Volume ◽  
Positive Control ◽  
Polymorphonuclear Neutrophil ◽  
Heat Stable ◽  
Dose Dependent

The aim of this study was to determine the regulatory role of pig seminal plasma in post-breeding uterine inflammation. Polymorphonuclear neutrophil (PMN) chemotaxis of lipopolysaccharide (LPS)-activated blood plasma or heat-inactivated blood plasma plus LPS containing increasing concentrations of seminal plasma was assessed in chemotactic chambers. Seminal plasma was diluted serially with McCoy's medium to concentrations of 50.0, 25.0, 12.5, 6.2 or 3.1% (v/v) and added to normal or heat-inactivated pig blood plasma that was activated with LPS before or after incubation in a 37 degrees C waterbath for 30 min. Chemotaxis was determined using blood-derived PMNs and was expressed as a percentage of the positive control of LPS-activated blood plasma. A linear dose-dependent suppression of chemotaxis by seminal plasma was observed for blood plasma activated before or after addition of seminal plasma. Compared with the positive control, concentrations of seminal plasma < 6.2% failed to suppress PMN chemotaxis (P < 0.05). A dose-dependent suppressive effect of seminal plasma on heat stable chemotactic components of pig blood plasma was also observed (P < 0.05). A marked suppression was observed at concentrations of seminal plasma > 12.5% of the sample volume (P < 0.05). These results indicate that seminal plasma suppresses chemotactic blood plasma components regardless of formation sequence (pre- or post-activation) or source (normal or heat-inactivated blood plasma). These results indicate that seminal plasma may be necessary in diluted boar semen used for artificial insemination to regulate post-breeding inflammation in sows.

14 EFFECT OF PORCINE SEMINAL PLASMA AND EGG YOLK ON CHEMOTAXIS AND PHAGOCYTOSIS OF NEUTROPHILS DERIVED FROM PERIPHERAL BLOOD OF PIGS AND COWS

2011 ◽  
Vol 23 (1) ◽  
pp. 113
Author(s):  
J.-C. Li ◽  
H. Funahashi
Keyword(s):  
Seminal Plasma ◽  
Peripheral Blood ◽  
Egg Yolk ◽  
Chemotactic Activity ◽  
Polymorphonuclear Neutrophil ◽  
Dependent Manner ◽  
Phagocytosis Assay ◽  
Concentration Dependent Manner ◽  
Phagocytotic Activity

The aim of this study was to determine the effects of porcine seminal plasma (0 to 20%, vol/vol) and egg yolk (0 to 20%, vol/vol) on chemotaxis and phagocytosis of porcine and bovine polymorphonuclear neutrophil (PMN) in vitro. Chemotaxis was determined using a blind well chamber. The phagocytosis assay was performed according to (Matthijs et al. 2000 J. Reprod. Fertil. 120, 265–273) with modification in pigs and cows. The serum-stimulated chemotactic activity of PMN (porcine, 1126.1 ± 14.4 cells/mm2 and bovine, 1067.1 ± 9.5 cells/mm2) was reduced (n = 4; P < 0.05) in the presence of 5% pig seminal plasma (pig, 1009.3 ± 12.4 cells/mm2 and cow, 800.0 ± 17.3 cells/mm2). More than 5% (vol/vol) of pig seminal plasma significantly decreased the chemotaxis of PMN in pigs and cows. The presence of 1% and higher concentrations (vol/vol) of seminal plasma reduced (n = 4; P < 0.05) leukocyte phagocytosis in pigs (37.1 ± 1.4 v. 41.7 ± 1.0%) and cows (36.4 ± 1.6 v. 42.1 ± 1.1%) compared with controls, and higher concentrations decreased the phagocytotic activity of PMNs in a concentration-dependent manner (n = 4; P < 0.05). Interestingly, 20% egg yolk increased (n = 4; P < 0.05) chemotaxis of PMN in pigs (953.5 ± 11.6 v. 789.9 ± 13.1 cells/mm2) and cow (988.6 ± 14.6 v. 790.4 ± 19.4 cells/mm2) compared with controls. The egg yolk increased (n = 4; P < 0.05) phagocytosis ability of porcine PMN (35.3 ± 1.6 v. 24.9 ± 2.4%) compared with the control. However, 20% egg yolk did not affect phagocytotic ability of PMN in cows (15.1 ± 1.6 v. 15.8 ± 1.0%). These results demonstrated that porcine seminal plasma, regardless of whether the species was pigs or cows, reduced chemotactic and phagocytotic activities of PMN. However, egg yolk significantly increased chemotactic activity of PMN in pigs and cows, while the egg yolk increased phagocytosis of PMN in pigs, but not in cows. In addition, porcine seminal plasma and caffeine reduced the egg yolk-induced increase in chemotactic and phagocytotic activities of PMN in both species. Supported by JSPS Grants-in-Aid (B20380154).

scholarly journals Seminal Plasma, Sperm Concentration, and Sperm-PMN Interaction in the Donkey: An In Vitro Model to Study Endometrial Inflammation at Post-Insemination

2020 ◽  
Vol 21 (10) ◽  
pp. 3478 ◽  
Author(s):  
Jordi Miró ◽  
Henar Marín ◽  
Jaime Catalán ◽  
Marion Papas ◽  
Sabrina Gacem ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Artificial Insemination ◽  
In Vitro Model ◽  
Sperm Concentration ◽  
Polymorphonuclear Neutrophils ◽  
Low Fertility ◽  
Fertility Rates ◽  
Vitro Model ◽  
Instrumental Role

In the donkey, artificial insemination (AI) with frozen-thawed semen is associated with low fertility rates, which could be partially augmented through adding seminal plasma (SP) and increasing sperm concentration. On the other hand, post-AI endometrial inflammation in the jenny is significantly higher than in the mare. While previous studies analyzed this response through recovering Polymorphonuclear Neutrophils (PMN) from uterine washings, successive lavages can detrimentally impact the endometrium, leading to fertility issues. For this reason, the first set of experiments in this work intended to set an in vitro model through harvesting PMN from the peripheral blood of jennies. Thereafter, how PMN, which require a triggering agent like formyl-methionyl-leucyl-phenylalanine (FMLP) to be activated, are affected by donkey semen was interrogated. Finally, we tested how four concentrations of spermatozoa (100 × 106, 200 × 106, 500 × 106 and 1000 × 106 spermatozoa/mL) affected their interaction with PMN. We observed that semen, which consists of sperm and SP, is able to activate PMN. Whereas there was a reduced percentage of spermatozoa phagocytosed by PMN, most remained attached on the PMN surface or into a surrounding halo. Spermatozoa not attached to PMN were viable, and most of those bound to PMN were also viable and showed high tail beating. Finally, only sperm concentrations higher than 500 × 106 spermatozoa/mL showed free sperm cells after 3 h of incubation, and percentages of spermatozoa not attached to PMN were higher at 3 h than at 1 h, exhibiting high motility. We can thus conclude that semen activates PMN in the donkey, and that the percentage of spermatozoa phagocytosed by PMN is low. Furthermore, because percentages of spermatozoa not attached to PMN were higher after 3 h than after 1 h of incubation, we suggest that PMN-sperm interaction plays an instrumental role in the reproductive strategy of the donkey.

scholarly journals Effects of components of semen extenders on the binding of stallion spermatozoa to bovine or equine zonae pellucidae

Reproduction ◽  
2012 ◽  
Vol 143 (5) ◽  
pp. 577-585 ◽  
Author(s):  
Marco A Coutinho da Silva ◽  
George E Seidel ◽  
Edward L Squires ◽  
James K Graham ◽  
Elaine M Carnevale
Keyword(s):  
Zona Pellucida ◽  
Binding Assay ◽  
Skim Milk ◽  
Milk Proteins ◽  
Sodium Caseinate ◽  
Sperm Binding ◽  
Threefold Increase ◽  
Series Of Experiments ◽  
Semen Extender ◽  
Positive Effect

The effects of semen extender components on the ability of stallion sperm to bind to the zona pellucida (ZP) and the suitability of using bovine ZP for a ZP-binding assay for stallion sperm were investigated in a series of experiments. In Experiment I, binding of stallion sperm to both bovine and equine ZP was significantly increased when a skim milk-based extender (EZM) was used. In Experiment II, a threefold increase in sperm binding to ZP was observed when sperm were diluted in EZM compared with diluents, which contained no milk (TALP, LAC, and EmCare). In Experiment III, centrifuging the sperm through Percoll did not increase sperm binding to the ZP but did remove any positive effect of EZM on sperm–ZP binding. In Experiment IV, exposure of either sperm or ZP to EZM before co-incubation did not increase sperm binding to ZP. In Experiment V, sperm diluted in TALP containing skim milk, EZM, or INRA96 bound more efficiently to the ZP than sperm diluted in TALP without milk proteins. In Experiment VI, sodium caseinate, native phosphocaseinate, and caseinoglycopeptide increased sperm binding to the ZP. In conclusion, diluents containing milk or milk proteins markedly enhanced the number of sperm bound to both equine and bovine ZP.

Inefficient in vitro killing of virulent or nonvirulent Salmonella typhimurium by murine polymorphonuclear neutrophils

1984 ◽  
Vol 30 (10) ◽  
pp. 1264-1270 ◽  
Author(s):  
Ellen Jo Baron ◽  
Richard A. Proctor
Keyword(s):  
Salmonella Typhimurium ◽  
Bactericidal Activity ◽  
In Vitro Studies ◽  
Polymorphonuclear Neutrophils ◽  
Polymorphonuclear Neutrophil ◽  
In Vitro System ◽  
Transient Decrease ◽  
In Vitro Incubation ◽  
Lag Period

The bactericidal capability of murine peritoneal polymorphonuclear neutrophils against virulent and nonvirulent Salmonella typhimurium was examined in an in vitro system. Although preincubation of the bacteria in specific murine antiserum elicited greater chemiluminescence from phagocytizing neutrophils than did incubation in normal murine serum, antiserum did not enhance ingestion, as less than 5% of the challenge was taken up by neutrophils under any of the conditions studied. Nonvirulent salmonellae showed a transient decrease in viable numbers early during in vitro incubation with or without intact neutrophils. Virulent salmonellae, however, were able to multiply without a lag period except when these bacteria were pretreated with antiserum and incubated in association with intact murine neutrophils. Results of these in vitro studies suggest that the murine polymorphonuclear neutrophil and antisalmonella antibody must act together to effect neutrophil-associated bactericidal activity against virulent salmonellae, and thus, that the neutrophil alone does not play a major role in the protection of unvaccinated, sensitive mice from disease caused by S. typhimurium.

Differences between antigenic determinants of pig and cat zona pellucida proteins

Reproduction ◽  
2000 ◽  
pp. 15-23 ◽  
Author(s):  
K Jewgenow ◽  
M Rohleder ◽  
I Wegner
Keyword(s):  
In Vitro Fertilization ◽  
Zona Pellucida ◽  
Antigenic Determinants ◽  
Vitro Fertilization ◽  
Contraceptive Vaccine ◽  
Sperm Binding ◽  
Dependent Inhibition ◽  
Dose Dependent Inhibition ◽  
Dose Dependent

Despite many efforts, the control of reproduction in feral cat populations is still a problem in urban regions around the world. Immunocontraception is a promising approach; thus the present study examined the suitability of the widely used pig zona pellucida proteins (pZP) for contraception in feral domestic cats. Purified zona pellucida proteins obtained from pig and cat ovaries were used to produce highly specific antisera in rabbits. Antibodies against pZP raised in rabbits or lions were not effective inhibitors of either in vitro sperm binding (cat spermatozoa to cat oocytes) or in vitro fertilization in cats, whereas antibodies against feline zona pellucida proteins (fZP) raised in rabbits showed a dose-dependent inhibition of in vitro fertilization. Immunoelectrophoresis, ELISA and immunohistology of ovaries confirmed these results, showing crossreactivity of anti-fZP sera to fZP and to a lesser extent to pZP, but no interaction of anti-pZP sera with fZP. It is concluded that cat and pig zonae pellucidae express a very small number of shared antigenic determinants, making the use of pZP vaccine in cats questionable. A contraceptive vaccine based on feline zona pellucida determinants will be a better choice for the control of reproduction in feral cats if immunogenity can be achieved.

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