Human Spermatozoa Quantitative Proteomic Signature Classifies Normo- and Asthenozoospermia

Abstract

Scarcely understood defects lead to asthenozoospermia, which results in poor fertility outcomes. Incomplete knowledge of these defects hinders the development of new therapies and reliance on interventional therapies, such as in vitro fertilization, increases. Sperm cells, being transcriptionally and translationally silent, necessitate the proteomic approach to study the sperm function. We have performed a differential proteomics analysis of human sperm and seminal plasma and identified and quantified 667 proteins in sperm and 429 proteins in seminal plasma data set, which were used for further analysis. Statistical and mathematical analysis combined with pathway analysis and self-organizing maps clustering and correlation was performed on the data set.It was found that sperm proteomic signature combined with statistical analysis as opposed to the seminal plasma proteomic signature can differentiate the normozoospermic versus the asthenozoospermic sperm samples. This is despite the results that some of the seminal plasma proteins have big fold changes among classes but they fall short of statistical significance. S-Plot of the sperm proteomic data set generated some high confidence targets, which might be implicated in sperm motility pathways. These proteins also had the area under the curve value of 0.9 or 1 in ROC curve analysis.Various pathways were either enriched in these proteomic data sets by pathway analysis or they were searched by their constituent proteins. Some of these pathways were axoneme activation and focal adhesion assembly, glycolysis, gluconeogenesis, cellular response to stress and nucleosome assembly among others. The mass spectrometric data is available via ProteomeXchange with identifier PXD004098. Molecular & Cellular

A quantitative global proteomics approach to understanding the functional pathways dysregulated in the spermatozoa of asthenozoospermic testicular cancer patients

“…Our bioinformatic results showed that the “cell movement of sperm” pathway was deactivated as the proteins associated with the sperm motility (AKAP4, ATP1A4, ATP2B4, GAPDHS, ROPN1, and SPAG6) were underexpressed in the asthenozoospermic TC patients (Table ). Earlier studies have also reported decreased expression of sperm motility associated proteins in the asthenozoospermic samples (Amaral et al ., ; Saraswat et al ., ). Functional analysis revealed ATP1A4 was involved in pathways associated with fertilization.…”

Characterization of the Sperm Proteome and Reproductive Outcomes with in Vitro, Fertilization after a Reduction in Male Ejaculatory Abstinence Period

“…Sperm Collection for Protein Modifications and Mass Spectrometry-The inclusion criteria were (1) normal semen parameters (concentration, total motility, and morphology) according to the WHO Laboratory Manual for the Examination and Processing of Human Semen (10); (2) no sexually transmitted diseases; (3) no drugs used in the past three months; and (4) a recent pregnancy (Ͻ2 years). Detailed sperm preparation protocols were established as previously described (11). Briefly, liquefied semen was centrifuged at 2000 ϫ g for 20 min at 4°C to separate spermatozoa from seminal plasma.…”

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

“…Asthenozoospermia is a common cause of male infertility, characterized by reduced sperm motility. Several comparative sperm proteomic studies have been conducted to delineate the proteins and associated pathways implicated in the molecular pathophysiology of asthenozoospermia [63][64][65][66]. The reduction of sperm motility has been recognized as a consequence of several factors, such as energy metabolism dysfunction, structural defects in sperm-tail protein components and differential expression of proteins involved in sperm motility such as cytochrome c oxidase subunit 6B (COX6B), outer dense fiber 2 (ODF) and tubulin beta 2B (TUBB2B) [20].…”

“…The reduction of sperm motility has been recognized as a consequence of several factors, such as energy metabolism dysfunction, structural defects in sperm-tail protein components and differential expression of proteins involved in sperm motility such as cytochrome c oxidase subunit 6B (COX6B), outer dense fiber 2 (ODF) and tubulin beta 2B (TUBB2B) [20]. Saraswat et al identified altered expression of proteins (see Table 1) associated with axoneme activation and focal adhesion assembly, glycolysis, gluconeogenesis, cellular response to stress and nucleosome assembly [64]. Comparative analysis of sperm proteome of normozoospermic and asthenozoospermic subjects by two-dimensional PAGE MALDI MS/MS resulted in the identification of eight proteins with altered expression [67].…”