Proteomic analysis of seminal extracellular vesicle proteins involved in asthenozoospermia by iTRAQ

Abstract

Asthenozoospermia is a common cause of male infertility, but in most cases its etiology is unknown. The exocytic cell vesicles called seminal extracellular vesicles in the human seminal fluid have been reported to play a pivotal role in promoting the motility of spermatozoa, and functional disorder of seminal extracellular vesicles may cause male infertility. To determine whether abnormal seminal extracellular vesicles are involved in asthenozoospermia, the differential abundance proteins between normozoospermic (NSEV) and asthenozoospermic seminal extracellular vesicles (ASEV) samples were analyzed by iTRAQ coupled with two‐dimensional liquid chromatography–tandem mass spectrometry. A total of 3,699 proteins were identified in the seminal extracellular vesicles (false discovery rate <0.01). Overall, 11 proteins were significantly upregulated (>1.2) in ASEV and 80 were significantly downregulated (<0.833). Functional bioinformatic analysis showed that these proteins with differential abundance were mainly associated with transport, metabolism, and signal pathways. The changes of OPTN, SMYD2, EIF2B2, TRPV6, ACE, PRSS8, and PPAP2A in ASEV were verified by western blot analysis, and we found that the abundance of TRPV6 markedly reduced in the seminal extracellular vesicles and ejaculated spermatozoa of asthenozoospermic patients, which indicated trpv6 was important in sperm motility. This study provides deeper insight into the involvement of seminal extracellular vesicles in asthenozoospermia and should aid the search for novel biomarkers of male infertility.

Proteomics of Extracellular Vesicles: Update on Their Composition, Biological Roles and Potential Use as Diagnostic Tools in Atherosclerotic Cardiovascular Diseases

“…A quantitative proteomic analysis based on a stable isotopic labelling method using isobaric tags for relative and absolute quantitation (iTRAQ) of urinary EV proteins was performed to discover potential biomarkers for prostate cancer diagnosis, and then the candidate proteins were verified using multiple reaction monitoring (MRM) MS [ 137 ]. Similarly, iTRAQ labelling was also used to quantify human seminal EV proteins [ 138 ]. A stable isotope labelling with amino acids in cell culture (SILAC) combined with high-resolution MS was also applied to perform a comprehensive analysis of proteins in microsatellite unstable colorectal cancer-derived EVs [ 139 ].…”

Exosome Composition and Seminal Plasma Proteome: A Promising Source of Biomarkers of Male Infertility

“…Although the transfer of proteins to SPZ represents a common feature of seminal exosomes, it is actually discussed whether distinct populations of vesicles carry out different tasks [ 22 , 23 ]. Whatever the case, the analysis of seminal proteome has corroborated the involvement of this fluid in the control of sperm physiology [ 24 ], motility [ 25 ], morphology [ 26 ], concentration [ 27 ] and protection from oxidation [ 28 ]. Interestingly, Ca 2+ signaling machinery required for sperm motility may be directly achieved from SP [ 29 ].…”

Seminal Plasma Proteome as an Indicator of Sperm Dysfunction and Low Sperm Motility in Chickens

“…Increasing evidence indicates that seminal plasma proteins are associated with a wide range of semen quality traits, including sperm motility (7,8), sperm morphology (9), sperm concentration (10), and sperm freezing endurance (11).…”

“…The Orbitrap mass spectrometer enables the analysis of highcomplexity samples like seminal plasma. In the present study, we performed label-free proteomics analysis based on SDS-PAGE fractionation of seminal Chicken Sperm Motility Related Seminal Plasma Proteins 8 plasma proteins obtained by Orbitrap instruments, aiming to reveal the seminal plasma proteomic profile of BJY roosters and identify the proteins associated with sperm motility. Through functional enrichment analyses, we have identified the biological functions of the seminal plasma proteins and suggested the altered proteins as potential candidate molecular regulators for sperm functions.…”