Colloid centrifugation removes seminal plasma and cholesterol from boar spermatozoa

2011 ◽  
Vol 23 (7) ◽  
pp. 858 ◽  
Author(s):  
R. Kruse ◽  
P. C. Dutta ◽  
J. M. Morrell
Keyword(s):  
Plasma Membrane ◽  
Seminal Plasma ◽  
Plasma Proteins ◽  
Single Layer ◽  
Sperm Cryopreservation ◽  
Before And After ◽  
Sperm Plasma Membrane ◽  
Seminal Plasma Proteins ◽  
Boar Spermatozoa ◽  
Layer Chromatography

The objective of the present study was to investigate the effect of Single-Layer Centrifugation (SLC) on boar spermatozoa, namely the effect of removal of seminal plasma proteins and cholesterol from the surface of spermatozoa. The presence of porcine seminal plasma proteins I and II (PSP-I/PSP-II) before and after SLC was studied using immunofluorescence, whereas the removal of cholesterol was shown qualitatively by thin-layer chromatography (TLC). Finally, the integrity of the sperm plasma membrane was observed by electron microscopy. It was shown that the seminal plasma proteins PSP-I and -II were removed from spermatozoa during SLC but could be restored by adding seminal plasma to the SLC-selected sperm samples. Some cholesterol was also lost from the spermatozoa during SLC but the plasma membrane itself appeared to be morphologically intact. Further studies are underway to examine the relevance of these findings to boar sperm cryopreservation and sperm fertility.

scholarly journals Single Layer Centrifugation Improves the Quality of Fresh Donkey Semen and Modifies the Sperm Ability to Interact with Polymorphonuclear Neutrophils

Animals ◽  
2020 ◽  
Vol 10 (11) ◽  
pp. 2128
Author(s):  
Marion Papas ◽  
Jaime Catalán ◽  
Sandra Recuero ◽  
Jane M. Morrell ◽  
Marc Yeste ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Membrane Potential ◽  
Mitochondrial Membrane Potential ◽  
Seminal Plasma ◽  
Plasma Proteins ◽  
Mitochondrial Membrane ◽  
Single Layer ◽  
Polymorphonuclear Neutrophils ◽  
Lipid Disorder ◽  
Seminal Plasma Proteins

This study sought to determine whether single layer centrifugation (SLC) of fresh donkey semen with Equicoll has any impact on sperm quality parameters and on the modulation of endometrial reaction following semen deposition using an in vitro model. Seventeen ejaculates from five jackasses were obtained using an artificial vagina and diluted in a skim-milk extender. Samples were either selected through SLC (Equicoll) or non-treated (control). Two experiments were performed. The first one consisted of incubating selected or non-selected spermatozoa at 38 °C for 180 min. Integrity and lipid disorder of sperm plasma membrane, mitochondrial membrane potential, and intracellular levels of calcium and reactive oxygen species were evaluated at 0, 60, 120, and 180 min. In the second experiment, polymorphonuclear neutrophils (PMN) isolated from jennies blood were mixed with selected and unselected spermatozoa. Interaction between spermatozoa and PMN was evaluated after 0, 60, 120, and 180 min of co-incubation at 38 °C. SLC-selection increased the proportions of spermatozoa with an intact plasma membrane and low lipid disorder, of spermatozoa with high mitochondrial membrane potential and with high calcium levels, and of progressively motile spermatozoa. In addition, selection through SLC augmented the proportion of phagocytosed spermatozoa, which supported the modulating role of seminal plasma proteins on sperm-PMN interaction. In conclusion, SLC of fresh donkey semen increases the proportions of functionally intact and motile spermatozoa, and appears to remove the seminal plasma proteins that inhibit sperm-PMN binding.

Adsorption of seminal plasma proteins by boar spermatozoa

1990 ◽  
Vol 34 (4) ◽  
pp. 691-700 ◽  
Author(s):  
K.W. Metz ◽  
Trish Berger ◽  
E.D. Clegg
Keyword(s):  
Seminal Plasma ◽  
Plasma Proteins ◽  
Seminal Plasma Proteins ◽  
Boar Spermatozoa

Seminal plasma proteins reduce protein tyrosine phosphorylation in the plasma membrane of cold-shocked ram spermatozoa

2002 ◽  
Vol 61 (2) ◽  
pp. 226-233 ◽  
Author(s):  
Rosaura Pérez-Pé ◽  
Patricia Grasa ◽  
Marta Fernández-Juan ◽  
Maria Luisa Peleato ◽  
José Álvaro Cebrián-Pérez ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Tyrosine Phosphorylation ◽  
Seminal Plasma ◽  
Plasma Proteins ◽  
Protein Tyrosine Phosphorylation ◽  
Protein Tyrosine ◽  
Seminal Plasma Proteins

Seminal plasma proteins inhibit in vitro- and cooling-induced capacitation in boar spermatozoa

2010 ◽  
Vol 22 (6) ◽  
pp. 893 ◽  
Author(s):  
Melissa L. Vadnais ◽  
Kenneth P. Roberts
Keyword(s):  
Mass Spectrometry ◽  
Seminal Plasma ◽  
Plasma Proteins ◽  
Acrosome Reaction ◽  
Heparin Binding ◽  
Total Proteins ◽  
Binding Properties ◽  
Seminal Plasma Proteins ◽  
Boar Spermatozoa

Dilute boar seminal plasma (SP) has been shown to inhibit in vitro capacitation and cooling-induced capacitation-like changes in boar spermatozoa, as assessed by the ability of the spermatozoa to undergo an ionophore-induced acrosome reaction. We hypothesised that the protein component of SP is responsible for this effect. To test this hypothesis, varying concentrations of total SP protein or SP proteins fractionated by heparin binding were assayed for their ability to inhibit in vitro capacitation, as well as cooling- and cryopreservation-induced capacitation-like changes. In vitro capacitation and cooling-induced capacitation-like changes were prevented by 10% whole SP, as well as by total proteins extracted from SP at concentrations greater than 500 μg mL−1. No amount of SP protein was able to prevent cryopreservation-induced capacitation-like changes. Total SP proteins were fractionated based on their heparin-binding properties and the heparin-binding fraction was shown to possess capacitation inhibitory activity at concentrations as low as 250 µg mL−1. The proteins in the heparin-binding fraction were subjected to mass spectrometry and identified. The predominant proteins were three members of the spermadhesin families, namely AQN-3, AQN-1 and AWN, and SP protein pB1. We conclude that one or more of these heparin-binding SP proteins is able to inhibit in vitro capacitation and cooling-induced capacitation-like changes, but not cryopreservation-induced capacitation-like changes, in boar spermatozoa.

scholarly journals Zinc-binding proteins from boar seminal plasma -- isolation, biochemical characteristics and influence on spermatozoa stored at 4°C.

2011 ◽  
Vol 58 (2) ◽  
Author(s):  
Marzena Mogielnicka-Brzozowska ◽  
Paweł Wysocki ◽  
Jerzy Strzeżek ◽  
Władysław Kordan
Keyword(s):  
Seminal Plasma ◽  
Plasma Proteins ◽  
Binding Proteins ◽  
Control Sample ◽  
Zinc Binding ◽  
Shielding Effect ◽  
Sperm Plasma Membrane ◽  
Native Electrophoresis ◽  
Seminal Plasma Proteins ◽  
Fast Flow

Affinity chromatography on Chelating Sepharose Fast Flow Gel-Zn(2+) was used for fractionation of boar seminal plasma proteins. Approximately 30% of total boar seminal plasma proteins showed affinity for zinc ions (ZnBP fraction). Native electrophoresis (PAGE) of ZnBP revealed six protein fractions which separated into 27 bands under denaturing conditions (SDS/PAGE). Two-dimensional electrophoresis (2D PAGE) showed 148 polypeptides with isoelectric points mostly in the basic and neutral pH range. The zinc-binding proteins comprise mainly 10-20 kDa polypeptides which are probably members of the spermadhesin family. ZnBP present in the incubation mixture of spermatozoa stored for 1 or 24 h at 4 °C allowed preservation of a higher percentage of cells exhibiting linear motility in comparison to a control sample stored in PBS. Presented results indicate that proteins binding Zn(2+) ions have a shielding effect on the sperm plasma membrane and acrosome of spermatozoa, protecting these structures against consequences of cold shock.

scholarly journals Seminal Plasma Proteins Prevent Detrimental Effects of Ram Sperm Cryopreservation and Enhance the Protective Effect of Lecithin

2017 ◽  
Vol 06 (02) ◽  
Author(s):  
SIgnacio Del Valle ◽  
Adriana Casao ◽  
Rosaura Perez Pe ◽  
William V Holt ◽  
Jose A Cebrian Perez ◽  
...  
Keyword(s):  
Protective Effect ◽  
Seminal Plasma ◽  
Plasma Proteins ◽  
Sperm Cryopreservation ◽  
Seminal Plasma Proteins ◽  
Ram Sperm

scholarly journals Difference of seminal plasma and sperm proteins in good and poor freezability boar ejaculates

2021 ◽  
Vol 53 (2) ◽  
Author(s):  
Janyaporn Rungruangsak ◽  
Junpen Suwimonteerabutr ◽  
Kakanang Buranaamnuay ◽  
Sariya Asawakarn ◽  
Naphat Chantavisoote ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Seminal Plasma ◽  
Plasma Proteins ◽  
Triosephosphate Isomerase ◽  
Progressive Motility ◽  
Sperm Proteins ◽  
Seminal Plasma Proteins ◽  
Boar Semen ◽  
High Level ◽  
Boar Spermatozoa

The present study was performed to compare the expression of sperm proteins, i.e. triosephosphate isomerase (TPI) and acrosin binding protein (ACRBP) and seminal plasma proteins, i.e. glutathione peroxidase 5 (GPX5) and fibronectin 1 (FN1), in boar semen with good, moderate and poor freezability. The study was conducted by determining the protein contents in 32 sperm samples and 38 seminal plasma samples of semen. The ejaculated semen was divided into two portions: the first portion was centrifuged to separate the pellet of sperm from the seminal plasma and the second portion was cryopreserved. After thawing, the ejaculates were classified into three groups according to their post-thawed sperm motility: good (60.2 ± 1.7%), moderate (29.3 ± 2.0%) and poor (16.6 ± 2.2%) freezabilities. The expressions of GPX5 and FN1 in seminal plasma and TPI and ACRBP in sperm were determined using Western blot analysis. It was found that, for sperm proteins, the level of TPI was negatively correlated with the post-thawed total sperm motility (r = -0.38, P = 0.029). For seminal plasma proteins, the level of FN1 in the seminal plasma was positively correlated with the post-thawed total sperm motility (r = 0.37, P = 0.021) and progressive motility (r = 0.39, P = 0.016). The expression of GPX5 was not correlated with any of the frozen–thawed sperm qualities (P > 0.05). In conclusions, boar semen containing a high level of FN1 in seminal plasma has better freezability. Frozen–thawed sperm motility was positively correlated with the level of FN1 in boar seminal plasma and negatively correlated with TPI in boar spermatozoa.

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