Seminal plasma proteins inhibit in vitro- and cooling-induced capacitation in 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.

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Characteristics of selected seminal plasma proteins and their application in the improvement of the reproductive processes in mammals

Polish Journal of Veterinary Sciences ◽

10.2478/v10181-011-0074-z ◽

2011 ◽

Vol 14 (3) ◽

pp. 489-499 ◽

Cited By ~ 17

Author(s):

M. Mogielnicka-Brzozowska ◽

W. Kordan

Keyword(s):

Seminal Plasma ◽

Plasma Proteins ◽

Acrosome Reaction ◽

Reproductive Tract ◽

Chromatin Condensation ◽

Female Reproductive Tract ◽

Sperm Chromatin ◽

Heparin Binding ◽

Seminal Plasma Proteins ◽

Reproductive Processes

Characteristics of selected seminal plasma proteins and their application in the improvement of the reproductive processes in mammals Understanding the biochemical processes associated with ovum fertilization and knowledge about the structure and function of individual substances participating in these processes is crucial for the development of biotechnological methods to improve reproduction of animals and humans. Among many components of seminal plasma, proteins and peptides play a specific role in regulation of the fertilization process, particularly through their ability to bind various types of ligands such as polysaccharides, lipids and ions. Heparin-binding proteins regulate capacitation and acrosome reaction processes. Affinity of plasma proteins to mannans of the fallopian tube epithelium facilitates formation of spermatozoa reservoirs in the female reproductive tract. Ability to bind phosphorylcholine is one of the conditions for the coating of the seminal plasma proteins on the sperm membrane and also determines the formation of oligomeric forms of certain proteins. Zinc binding by seminal plasma proteins regulates sperm chromatin condensation state. It also affects motility of these cells and acrosome reaction. The interspecies analysis indicates significant structural and functional similarities, especially for the proteins with low molecular weight. Fertility associated proteins (FAPs) have been determined in the bull, stallion, boar, ram and dog. The contents of these proteins correlate with the indicators of the fertilizing abilities of sperm. In humans, several seminal plasma proteins were found which serve as diagnostic markers of spermatogenesis, seminiferous epithelium state, and azoospermia. To determine the semen ability for preservation, measurement of some seminal plasma protein content may also be used. Addition of specific plasma proteins to a spermatozoa solution undergoing the process of preservation may be used to retain the features of the cells responsible for efficient fertilization.

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Supplementation of Seminal Plasma-Heparin Binding Proteins to Capacitation Medium Increases In Vitro Acrosome Reaction Percentage in Beetal Bucks

Theriogenology Insight - An International Journal of Reproduction in all Animals ◽

10.30954/2277-3371.01.2020.2 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Navjot Singh Dhillon

Keyword(s):

Seminal Plasma ◽

Acrosome Reaction ◽

Binding Proteins ◽

Heparin Binding ◽

Plasma Heparin

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The Ubiquitin-Proteasome System Does Not Regulate the Degradation of Porcine β-Microseminoprotein during Sperm Capacitation

International Journal of Molecular Sciences ◽

10.3390/ijms21114151 ◽

2020 ◽

Vol 21 (11) ◽

pp. 4151

Author(s):

Lucie Tumova ◽

Michal Zigo ◽

Peter Sutovsky ◽

Marketa Sedmikova ◽

Pavla Postlerova

Keyword(s):

Seminal Plasma ◽

Plasma Proteins ◽

Protein Composition ◽

Surface Protein ◽

Ubiquitin Proteasome System ◽

Sperm Capacitation ◽

Sperm Surface ◽

Ubiquitin Proteasome ◽

Seminal Plasma Proteins

Sperm capacitation, one of the key events during successful fertilization, is associated with extensive structural and functional sperm remodeling, beginning with the modification of protein composition within the sperm plasma membrane. The ubiquitin-proteasome system (UPS), a multiprotein complex responsible for protein degradation and turnover, participates in capacitation events. Previous studies showed that capacitation-induced shedding of the seminal plasma proteins such as SPINK2, AQN1, and DQH from the sperm surface is regulated by UPS. Alterations in the sperm surface protein composition also relate to the porcine β-microseminoprotein (MSMB/PSP94), seminal plasma protein known as immunoglobulin-binding factor, and motility inhibitor. MSMB was detected in the acrosomal region as well as the flagellum of ejaculated boar spermatozoa, while the signal disappeared from the acrosomal region after in vitro capacitation (IVC). The involvement of UPS in the MSMB degradation during sperm IVC was studied using proteasomal interference and ubiquitin-activating enzyme (E1) inhibiting conditions by image-based flow cytometry and Western blot detection. Our results showed no accumulation of porcine MSMB either under proteasomal inhibition or under E1 inhibiting conditions. In addition, the immunoprecipitation study did not detect any ubiquitination of sperm MSMB nor was MSMB detected in the affinity-purified fraction containing ubiquitinated sperm proteins. Based on our results, we conclude that UPS does not appear to be the regulatory mechanism in the case of MSMB and opening new questions for further studies. Thus, the capacitation-induced processing of seminal plasma proteins on the sperm surface may be more complex than previously thought, employing multiple proteolytic systems in a non-redundant manner.

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Adsorption of seminal plasma proteins by boar spermatozoa

Theriogenology ◽

10.1016/0093-691x(90)90024-n ◽

1990 ◽

Vol 34 (4) ◽

pp. 691-700 ◽

Cited By ~ 26

Author(s):

K.W. Metz ◽

Trish Berger ◽

E.D. Clegg

Keyword(s):

Seminal Plasma ◽

Plasma Proteins ◽

Seminal Plasma Proteins ◽

Boar Spermatozoa

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The importance of bovine seminal plasma proteins in the sperm function and fertility

SPERMOVA ◽

10.18548/aspe/0009.12 ◽

2021 ◽

Vol 11 (2) ◽

pp. 83-95

Author(s):

María Alejandra Cardozo ◽

◽

Jaime Antonio Cardozo ◽

Fabian Rueda

Keyword(s):

Seminal Plasma ◽

Plasma Proteins ◽

Acrosome Reaction ◽

Protein Identification ◽

Seminal Plasma Proteins ◽

Biotechnological Processes ◽

And Oxidative Stress ◽

Social Sectors ◽

Sperm Functions

Bovine livestock is one of the most important economic and social sectors for many countries. In this sense, the development of strategies to improve reproductive bull fertility and reproduction rates is relevant. It's highlighted the role of seminal plasma proteins (SPP) in reproductive fertility, so it has found close relationships among studies on the structure and biological activity of SPP, with seminal quality, including viability, sperm motility, and morphology. In addition, they have been found to regulate sperm functions such as capacitation, acrosome reaction, and they are even related to protecting sperm against thermal and oxidative stress. Moreover, the methods of separation and protein identification and their contribution to characterizing the bovine SP proteome should be also highlighted. In this sense, the most recent studies have been directed towards developing supplements with SPP that improve quality sperm subjected to cryopreservation processes. Research has begun and should forward to establish how the networks or sets of proteins are related to the functioning and fertility of sperm, the search for biomarkers of fertility, and the use of proteins in biotechnological processes, to increase efficiency reproductive.

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

Biology of Reproduction ◽

10.1093/biolre/ioab225 ◽

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.

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Ram seminal plasma and its functional proteomic assessment

Reproduction ◽

10.1530/rep-18-0627 ◽

2019 ◽

Vol 157 (6) ◽

pp. R243-R256 ◽

Cited By ~ 12

Author(s):

T Leahy ◽

J P Rickard ◽

N C Bernecic ◽

X Druart ◽

S P de Graaf

Keyword(s):

Seminal Plasma ◽

Plasma Proteins ◽

Reproductive Tract ◽

Reproductive Technologies ◽

Female Reproductive Tract ◽

Sperm Function ◽

Seminal Plasma Proteins ◽

Ram Sperm

Ejaculation results in the confluence of epididymal spermatozoa with secretions of the accessory sex glands. This interaction is not a prerequisite for fertilisation success, but seminal factors do play a crucial role in prolonging the survival of spermatozoa bothin vitroandin vivoby affording protection from handling induced stress and some selective mechanisms of the female reproductive tract. Reproductive biologists have long sought to identify specific factors in seminal plasma that influence sperm function and fertility in these contexts. Many seminal plasma proteins have been identified as diagnostic predictors of sperm function and have been isolated and appliedin vitroto prevent sperm damage associated with the application of artificial reproductive technologies. Proteomic assessment of the spermatozoon, and its surroundings, has provided considerable advances towards these goals and allowed for greater understanding of their physiological function. In this review, the importance of seminal plasma will be examined through a proteomic lens to provide comprehensive analysis of the ram seminal proteome and detail the use of proteomic studies that correlate seminal plasma proteins with ram sperm function and preservation ability.

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Aggregated Forms of Bull Seminal Plasma Proteins and Their Heparin-Binding Activity

Collection of Czechoslovak Chemical Communications ◽

10.1135/cccc20040616 ◽

2004 ◽

Vol 69 (3) ◽

pp. 616-630 ◽

Cited By ~ 2

Author(s):

Petra Jelínková ◽

Helena Ryšlavá ◽

Jiří Liberda ◽

Věra Jonáková ◽

Marie Tichá

Keyword(s):

Molecular Weight ◽

Seminal Plasma ◽

Plasma Proteins ◽

Sodium Dodecyl ◽

Reversed Phase ◽

Binding Activity ◽

Size Exclusion ◽

Heparin Binding ◽

Aggregation State ◽

Seminal Plasma Proteins

Heparin-binding activity of bull seminal plasma proteins was shown to be dependent on their aggregation state. The protein fraction interacting with immobilized heparin was characterized by large polydispersity in the region of molecular weight of 60 000-10 000, while that not retained on the affinity carrier was present as aggregates with molecular weight >100 000. Components of heparin-binding and non-heparin-binding fractions were separated by RP HPLC (reversed-phase HPLC) and analyzed by SDS (sodium dodecyl sulfate) electrophoresis and N-terminal sequencing. Size exclusion chromatography of whole seminal plasma and heparin-binding proteins in the presence of D-fructose (as a component of seminal plasma) showed that the region of molecular weights of protein-associated forms was shifted to lower values. An increase of heparin-binding activity of bull proteins, as determined by ELBA (Enzyme-Linked Binding Assay), correlates with a decrease of their aggregation state. The modulation of the aggregation state of bull proteins by seminal plasma components and, in this way, also of their heparin-binding properties suggests possible mechanisms for capacitation mediated by these proteins.

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Electrophoretic profile of seminal proteins and their correlation with in vitro sperm characters in Black Bengal buck semen

Veterinary World ◽

10.14202/vetworld.2019.621-628 ◽

2019 ◽

Vol 12 (5) ◽

pp. 621-628 ◽

Cited By ~ 5

Author(s):

M. Karunakaran ◽

Vivek C. Gajare ◽

Ajoy Mandal ◽

Mohan Mondal ◽

S. K. Das ◽

...

Keyword(s):

Molecular Weight ◽

Seminal Plasma ◽

Plasma Proteins ◽

Membrane Integrity ◽

Sperm Proteins ◽

Sds Page ◽

Sperm Characters ◽

Functional Membrane ◽

Seminal Plasma Proteins

Aim: This study aimed to study the electrophoretic properties of seminal plasma and sperm proteins of Black Bengal buck semen and their correlation with in vitro sperm characters and freezability. Materials and Methods: Semen ejaculates from nine Black Bengal bucks were collected by artificial vagina (n=20/buck). Ejaculates were evaluated for in vitro sperm characters and electrophoretic profile of seminal protein. In vitro sperm characters were evaluated immediately after collection, after completion of equilibration period, and after freeze-thawing. For seminal protein studies, seminal plasma proteins were precipitated by ice-cold ethanol method, and sperm proteins were extracted by Triton X detergent extraction method. Discontinuous sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was performed to assess the molecular weight of seminal proteins. Correlation between in vitro sperm characters and protein bands was determined by Pearson's correlation coefficient, and two-way ANOVA was applied to find the individual buck differences. Results: Significant difference (p<0.01) among the bucks was noticed in the in vitro sperm characters evaluated at all the three stages of semen evaluation such as immediately after collection, after completion of equilibration period, and post-freeze thawing. Progressive loss of sperm motility, membrane integrity, and other in vitro sperm characters were noticed during cryopreservation. A total of ten protein bands in the molecular weight ranging from 17 to 180 kDa were found in the SDS-PAGE of seminal plasma proteins, while nine bands of 17-134 kDa were observed in sperm proteins. Seminal plasma proteins of molecular weight 75, 62-49, 20, and 17 kDa and sperm proteins of 75, 20, and 17 kDa were present in all the nine bucks (100%) screened, and variation among the bucks was noticed for the presence of other proteins. Seminal plasma protein of 180-134 kDa showed a negative correlation with individual motility (−0.716) and functional membrane integrity of sperm cells (−0.724) in post-freeze-thaw analysis and 48 kDa protein had a positive correlation with individual motility (0.649) and functional membrane integrity of sperm cells (0.664) in post-thaw analysis. Sperm proteins of 63 kDa had a negative correlation (−0.616) with sperm concentration in neat semen. Conclusion: Variation among the bucks was noticed in the in vitro sperm characters and semen freezability. Correlation between seminal proteins and in vitro sperm characters and semen freezability had been found which might be useful as a tool to select breeding bucks.

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