Equine seminal plasma and sperm membrane: Functional proteomic assessment.
Abstract: During ejaculation, a large amount of seminal plasma proteins interact with the sperm membrane, leading to a series of biochemical and structural changes implicated in sperm function and gamete interaction. However, the roles of the majority of these proteins remain unknown. This study aimed to investigate the proteome and functionality of the major equine proteins of seminal plasma and the sperm membrane. Seminal plasma and enriched-membrane proteins (150 μg) were separated by two-dimensional gel electrophoresis, and the respective maps were analyzed. Protein identification was performed by in-gel digestion and tandem mass spectrometry (GeLC-MS/MS). Samples were also submitted to in-solution digestion (complex protein mixture) and identified by shotgun analysis by LC-MS/MS; bioinformatic tools were used to investigate protein functions. Seminal plasma and sperm membrane extract maps contained 91.0 ± 8.2 spots and 245.3 ± 11.3 spots, respectively, within the 3-10 pH range. In total, the most abundant proteins identified in 2D maps and in complex protein mixtures included 24 proteins for seminal plasma and 33 for sperm membrane extract, with a high degree of confidence (P < 0.05). Of these, HSP1, CRISP3 and KLK1E2 were the most abundant in seminal plasma; HSP1 was highly abundant in sperm membrane extract, in many isoforms, which is related to membrane destabilization and may compromise sperm preservation. HSP1-polybromo-1 interactions suggested a role in DNA stabilization. Prosaposin was identified in seminal plasma and may play a role in the fertilization process. IZUMO4, a member of the IgSF family involved in the prefertilization stages, was identified in 2D gel and MS/MS analysis of sperm membrane extract. Ten proteins of seminal plasma were found to interact with the sperm membrane and were related to binding and catalytic activities (clusterin, CRISP3, epididymal sperm-binding protein 1, kallikrein1E2, seminal plasma protein A3, and HSP1). Additionally, other identified proteins were associated with DNA integrity, capacitation and recognition of pregnancy. These findings indicate that the binding of specific proteins to the plasma membrane during ejaculation may influence sperm survival after cryopreservation and may play a role in decreasing the quality in stallions with toxic seminal plasma. Elucidation of these interactions is an important step in understanding the biological processes related to equine fertility and facilitates future investigations on the selection and application of low freezability semen strategies.
Copyright © 2020 Elsevier Inc. All rights reserved.
Publication Date: 2020-06-14 PubMed ID: 32679458DOI: 10.1016/j.theriogenology.2020.06.014Google Scholar: Lookup
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- Journal Article
Summary
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This research investigates the role of proteins in horse seminal plasma and the sperm membrane, focusing particularly on how these proteins interact and potentially impact sperm function and the process of fertilization.
Proteomic Assessment Method
- The researchers conducted a proteomic assessment of seminal plasma and the sperm membrane of equine.
- Proteins from the seminal plasma and the sperm membrane were separated using a technique known as two-dimensional gel electrophoresis. This allowed them to create a map of the proteins.
- The protein identification involved in-gel digestion and tandem mass spectrometry (GeLC-MS/MS), this approach can identify and quantify proteins clearly and specifically.
- Additionally, samples were submitted to in-solution digestion and identified by an analysis known as shotgun by LC-MS/MS.
- The functionality and roles of these proteins were further investigated with the help of bioinformatics tools.
Findings of the Study
- In the 3-10 pH range, the researchers identified approximately 91.0 ± 8.2 spots in the seminal plasma and 245.3 ± 11.3 spots in the sperm membrane extract map.
- 24 proteins for seminal plasma and 33 for sperm membrane extract were identified to be the most abundant.
- Hsp1 was highly abundant both in seminal plasma and in the sperm membrane extract. This protein was linked to membrane destabilization and might influence the sperm preservation.
- A protein named Prosaposin identified in the seminal plasma is suggested to possibly play a role in the fertilization process.
- IZUMO4, another protein that was identified during the analysis, belonging to the IgSF family, is involved in prefertilization stages.
- Furthermore, the researchers identified that ten proteins from the seminal plasma interact with the sperm membrane. Notably, these proteins were related to binding and catalytic activities which include clusterin, CRISP3, epididymal sperm-binding protein 1, kallikrein1E2, seminal plasma protein A3, and HSP1.
- Additional proteins were found to be associated with DNA integrity, capacitation, and recognition of pregnancy.
Affect on Equine Fertility
- The study concluded that these interactions between specific proteins and the plasma membrane during ejaculation can potentially influence sperm survival and how it reacts to cryopreservation. Essentially, these proteins may play a crucial role in the quality of the sperm, particularly in cases of stallions with toxic seminal plasma.
- Understanding these interactions is crucial as it aids in comprehending the biological processes linked to equine fertility, offering guidance for future research and potential interventions regarding low freezability semen strategies.
Cite This Article
APA
Guasti PN, Souza FF, Scott C, Papa PM, Camargo LS, Schmith RA, Monteiro GA, Hartwig FP, Papa FO.
(2020).
Equine seminal plasma and sperm membrane: Functional proteomic assessment.
Theriogenology, 156, 70-81.
https://doi.org/10.1016/j.theriogenology.2020.06.014 Publication
Researcher Affiliations
- Department of Veterinary Surgery and Animal Reproduction, School of Veterinary Medicine and Animal Science, FMVZ, São Paulo State University (UNESP), Botucatu, SP, Brazil.
- Department of Veterinary Surgery and Animal Reproduction, School of Veterinary Medicine and Animal Science, FMVZ, São Paulo State University (UNESP), Botucatu, SP, Brazil.
- Department of Veterinary Surgery and Animal Reproduction, School of Veterinary Medicine and Animal Science, FMVZ, São Paulo State University (UNESP), Botucatu, SP, Brazil.
- Department of Veterinary Surgery and Animal Reproduction, School of Veterinary Medicine and Animal Science, FMVZ, São Paulo State University (UNESP), Botucatu, SP, Brazil.
- Department of Veterinary Surgery and Animal Reproduction, School of Veterinary Medicine and Animal Science, FMVZ, São Paulo State University (UNESP), Botucatu, SP, Brazil.
- Department of Veterinary Surgery and Animal Reproduction, School of Veterinary Medicine and Animal Science, FMVZ, São Paulo State University (UNESP), Botucatu, SP, Brazil.
- Department of Veterinary Clinic and Surgery, School of Veterinary Medicine, Federal University of Minas Gerais (UFMG), Belo Horizonte, Brazil.
- Department of Veterinary Surgery and Animal Reproduction, School of Veterinary Medicine and Animal Science, FMVZ, São Paulo State University (UNESP), Botucatu, SP, Brazil.
- Department of Veterinary Surgery and Animal Reproduction, School of Veterinary Medicine and Animal Science, FMVZ, São Paulo State University (UNESP), Botucatu, SP, Brazil. Electronic address: papa@fmvz.unesp.br.
MeSH Terms
- Animals
- Chromatography, Liquid / veterinary
- Female
- Horses
- Male
- Pregnancy
- Proteomics
- Semen
- Seminal Plasma Proteins
- Spermatozoa
- Tandem Mass Spectrometry / veterinary
Conflict of Interest Statement
Declaration of competing interest The authors state that they have no conflicts of interest to declare.
Citations
This article has been cited 11 times.- Gouletsou PG, Tsangaris GT, Katsarou EI, Bourganou MV, Barbagianni MS, Venianaki AP, Bouroutzika E, Anagnostopoulos AK, Fthenakis GC, Katsafadou AI. Proteomics Evaluation of Semen of Clinically Healthy Beagle-Breed Dogs. Vet Sci 2022 Dec 15;9(12).
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- Zhang M, Bromfield EG, Helms JB, Gadella BM. The fate of porcine sperm CRISP2 from the perinuclear theca before and after in vitro fertilization†. Biol Reprod 2022 Nov 14;107(5):1242-1253.
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- Said S, Maulana T, Iskandar H, Kaiin EM, Khaerunnisa I, Putra WPB, Hasan F, Arifiantini RI. Sperm protein profile and their correlation with frozen semen quality of indigenous Indonesian buffalo bulls. J Adv Vet Anim Res 2024 Dec;11(4):846-855.
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