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Sperm and Seminal Plasma Proteomics: Molecular Changes Associated with Varicocele-Mediated Male Infertility

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Sperm and Seminal Plasma Proteomics: Molecular Changes Associated with Varicocele-Mediated Male Infertility Review Article Male reproductive health and infertility pISSN: 2287-4208 / eISSN: 2287-4690 World J Mens Health Published online Jul 26, 2019 https://doi.org/10.5534/wjmh.190018 Sperm and Seminal Plasma Proteomics: Molecular Changes Associated with Varicocele-Mediated Male Infertility Manesh Kumar Panner Selvam , Ashok Agarwal American Center for Reproductive Medicine, Cleveland Clinic, Cleveland, OH, USA Male infertility is a rising problem and the etiology at the molecular level is unclear. Use of omics has provided an insight into the underlying cellular changes in the spermatozoa of infertile men. Proteomics is one the promising omics techniques for biomarker screening that can provide complete information on molecular processes associated with male infertility. Vari- cocele is a pressing issue in the field of male infertility and the search for an appropriate diagnostic and therapeutic biomark- er is still ongoing. In this review, we discuss the reports on proteomic profiles of sperm and seminal plasma in male infertility and provide an in-depth insight into varicocele studies associated with male infertility. Keywords: Proteomics; Seminal plasma; Sperm; Varicocele This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. INTRODUCTION metabolites and endocrine factors are associated with varicocele pathophysiology (Fig. 1). The mechanisms Currently, infertility is one of the top most concerns associated with the pathophysiology of varicocele have related to male reproductive dysfunction and male been reviewed in detail by Agarwal et al [7] and Cho et infertility contributes to nearly 50% of overall infertil- al [6]. Furthermore, varicocele drastically alters semen ity cases [1]. The incidence of varicocele is about 21% to parameters [8,9] and these patients exhibit compro- 41% in men with primary infertility and 75% to 80% mised fertility [10]. in men with secondary infertility [2,3]. Varicocele is Laboratory evaluation of male infertility in varico- characterized by the enlargement of the pampiniform cele patients is based on basic semen analysis (sperm plexus, which may be due to the presence of malfunc- concentration, motility, vitality, and morphology) as tioning valves. In these patients, testicular function per the World Health Organization 2010 guidelines [11]. is affected due to the retrograde flow of blood. Thus, Additionally, advanced laboratory tests such as those varicocele has a detrimental effect on spermatogenesis for the quantification of reactive oxygen species (ROS) by inducing a state of testicular hyperthermia, hypoxia [12] and antioxidants in semen [12], oxidation–reduction and oxidative stress [4-6]. In addition, the reflux of potential in ejaculated semen by Male Infertility Oxi- Received: Jan 28, 2019 Revised: Jun 3, 2019 Accepted: Jun 18, 2019 Published online Jul 26, 2019 Correspondence to: Ashok Agarwal https://orcid.org/0000-0003-0585-1026 American Center for Reproductive Medicine, Cleveland Clinic, Mail Code X-11, 10681 Carnegie Avenue, Cleveland, OH 44195, USA. Tel: +1-216 444-9485, Fax: +1-216 445-6049, E-mail: [email protected], Website: CCF.org/ReproductiveResearchCenter Copyright © 2019 Korean Society for Sexual Medicine and Andrology https://doi.org/10.5534/wjmh.190018 Varicocele Endocrine Testicular Scrotal Reflux of Cadmium factor hypoxia hyperthemia metabolites accumulation Impaired spermatogenesis Elevated ROS Sperm membrane Sperm DNA Apoptosis of lipid peroxidation fragmentation spermatozoa Impaired sperm function Fig. 1. Pathophysiology of varicocele. CCF Decreased male reproductive potential C 2018 ROS: reactive oxygen species. dative System (MiOXSYS®; Aytu BioScience Inc., En- mis and accessory sex glands. Seminal plasma proteins glewood, CO, USA) [13], and sperm DNA fragmentation interact with and modulate sperm functions, such as (SDF) assessment by terminal deoxynucleotidyl trans- capacitation, hyperactivation and acrosome reaction ferase-mediated dUTP nick-end labelling (TUNEL) as- required for fertilization [17,23]. In varicocele condition, say [13] and sperm chromatin structure assay [14] are these sperm functions are compromised, and sperm/ used to identify the functionality of spermatozoa for seminal plasma proteomic analysis have reported al- further utilization in assisted reproductive technology. tered expression of sperm and seminal plasma proteins However, these tests lack the information on molecular [24-26]. changes at the subcellular level associated with the In this review, we discuss the proteins involved in poor fertilizing ability of spermatozoa. the regulation of sperm functions that are present in Advancement in the current omics techniques, espe- both the cellular (sperm) and fluidic component (semi- cially proteomics have revolutionized the field of sperm nal plasma) of semen. In addition, we have discussed molecular biology. Proteomics, a high throughput the future of proteomics as a potential clinical tool for platform, is used to identify and select non-invasive the diagnosis and management of varicocele patients. biomarkers for the diagnosis of male infertility [15-17]. This has facilitated the identification of cellular and BACKGROUND OF SPERM AND molecular pathways that are being dysregulated in the SEMINAL FLUID PROTEOMICS spermatozoa of infertile patients [18]. Post-translational modification (PTM) of sperm proteins provide valuable Semen is a highly complex biofluid that contains information pertaining to the biomolecules associated proteins and peptides with varied functions. The main with the fertilization potential of spermatozoa [19,20]. components of semen include spermatozoa (cellular) Altered expression of sperm proteins in infertile pa- and seminal fluid (enriched with proteins). Sperm tients indicate compromised spermatogenesis or defects proteomics came into the limelight because of the in vital sperm functions, such as capacitation, hyper- transcriptionally and translationally inert property activation and acrosome reaction, which are essential of spermatozoa [27]. Characterization of proteins in for the fertilization process [21,22]. Another important spermatozoa and seminal plasma provides insight into component of semen is seminal plasma, which is con- the functions of specific proteins related to fertility stituted by the secretions derived from testes, epididy- [28]. Sperm proteins regulate the molecular pathways 2 www.wjmh.org Manesh Kumar Panner Selvam and Ashok Agarwal: Proteomics of Varicocele-Mediated Male Infertility such as protein and energy metabolism, PTMs, DNA 10% of the seminal plasma proteins are contributed by damage and oxidative stress response [19,29,30]. Sper- seminal extracellular vesicles (including epididymo- matogenesis involves complex processes that ultimately somes and prostasomes) [15,32]. So far, 2,064 proteins produce the male gamete with specialized functions. have been identified in the seminal plasma [32]. About The developmental process of spermatozoa is regulated 70% of these proteins were also identified in sperma- by protein–protein interaction [31]. Amaral et al [30] tozoa [29,36]. Altered expressions of seminal plasma identified 6,198 proteins in spermatozoa and 30% of proteins have a direct effect on spermatozoa homeo- these proteins originate from the testis. Protein charac- stasis and functions. Comparative proteomic approach terization studies revealed that a total of 898, 984, and allows the identification of the underlying molecular 532 proteins were present in the sperm head, tail and causes associated with the pathology of spermatozoa. both locations, respectively [32]. Protein profiling using Expression of semen proteins varies from one condi- 2-dimensional (2D)-gel matrix-assisted laser desorption/ tion to another. Proteomics analysis demonstrated the ionization time-of-flight (MALDI-TOF) detected that differential expression of proteins in semen and its the sperm proteins were distributed in cytoplasm (37%), potential use as non-invasive biomarkers in infertile mitochondria (19%), nuclear (5%), tail and flagella (3%), men with abnormal semen parameters. In azoospermic and acrosome (2%) [33]. For the first time in human men, the proteins ACPP (prostatic acid phosphatase), spermatozoa, 27 proteins present in the 26S proteasome KLK3 (prostate-specific antigen, PSA), CLU (clusterin), complex was mapped from 1,760 proteins using 1-di- AZGP1 (zinc-alpha-2-glycoprotein), and PAEP (glycode- mensional (1D)-sodium dodecyl sulfate-polyacrylamide lin) were absent in seminal plasma [37,38]. Drabovich et gel electrophoresis (SDS-PAGE) combined with GeLC- al [39] validated TEX101 (testis-expressed protein 101) tandem mass spectrometry (MS/MS) technique [34]. as a biomarker in azoospermia and ECM1 (extracel- Bioinformatic tools predicted that the biological path- lular matrix protein 1) to distinguish non-obstructive ways, such as oxidative phosphorylation and glycolysis azoospermia from vasectomy. In the case of astheno- are influenced by sperm proteins [18,29,30]. zoospermia, PTPN14 (tyrosine-protein phosphatase non- Apart from sperm proteins, seminal plasma pro- receptor type 14) was dysregulated [40], whereas CST3 teins are also essential for the maintenance of sperm (cystatin-C) was downregulated, and KLK3 and SEMG1 functionality [35]. Seminal plasma
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