Comparative Proteomics Reveal the Association between SPANX Proteins and Clinical Outcomes of Artificial Insemination with Donor Sperm

Abstract

Semen analysis is used for diagnosing male infertility and evaluating male fertility for more than a century. However, the semen analysis simply represents the population characteristics of sperm. It is not a comprehensive assessment of the male reproductive potential. In this study, 20 semen samples from human sperm bank with distinctive artificial insemination with donor sperm (AID) clinical outcomes were collected and analyzed using a two-dimensional differential in-gel electrophoresis (2D-DIGE); 45 differentially expressed protein spots were obtained, and 26 proteins were identified. Most differentially expressed proteins were related to sperm motility, energy consumption, and structure. These identified proteins included several sperm proteins associated with the nucleus on the X chromosome (SPANX) proteins. This prospective study aimed to investigate the association between the expression levels of SPANX proteins and the AID clinical outcomes. The proteins identified in this study provided a reference for the molecular mechanism of sperm fertility and revealed a predictive value of the SPANX proteins.

Proteomic Analyses of Human Sperm Cells: Understanding the Role of Proteins and Molecular Pathways Affecting Male Reproductive Health

“…Few studies have proposed that the use of sperm with round cells in the protein extraction process may contaminate sperm proteome. Separation of round cells and leukocytes are carried out using gradient centrifugation method and the pure fraction of the spermatozoa are used for proteomic analysis [21,[33][34][35][36][37]. Recently, Panner Selvam et al investigated the interference of round cell proteins in the proteome of sperm and their effect on biological pathways associated with sperm function [38,39].…”

Proteomics and metabolomics — Current and future perspectives in clinical andrology

“…Semen samples are subjected to different types of processing techniques such as single gradient centrifugation, double gradient centrifugation or multilayer gradient centrifugation to separate the spermatozoa from the seminal plasma. Several studies have used a density gradient centrifugation step to remove round cells and immature germ cells from semen samples to isolate a pure sperm fraction (Amaral, Paiva, et al., 2014; Intasqui et al., 2013; Martínez‐Heredia et al., 2006; Wang et al., 2014, 2018). Further, proteomic studies on round cells and spermatozoa revealed that the influence of round cell proteins as contaminant in the sperm proteome was insignificant or very negligible (Panner Selvam, Agarwal, Dias et al., 2019; Panner Selvam, Agarwal, Dias, Martins, & Samanta, 2019).…”

Presence of Round Cells Proteins do not Interfere with Identification of Human Sperm Proteins from Frozen Semen Samples by LC-MS/MS

“…Gradient centrifugation techniques are widely used to process the semen samples to obtain a pure population of spermatozoa. To date, some studies have used only pure cultures of sperm (without round cells) for global proteomic analysis [17,18,19,20,21], whereas others have used neat semen samples [20,22,23,24]. Thus, it is still unclear (1) whether the processing of semen samples have any interference in the sperm protein expression, and (2) what the contribution is of non-spermatogenic proteins in the evaluation of sperm proteomic profile.…”