Differential motility parameters and identification of proteomic profiles of human sperm cryopreserved with cryostraw and cryovial

Abstract

Background Although sperm cryopreservation has been widely used in human reproductive medicine as an integral infertility management in infertility clinics and for banking sperm in sperm banks, the freezing/thawing protocols are not optimal. The freezing and thawing processes result in changes at both structural and molecular levels, some even detrimental, in human sperm when compared with fresh sperm. The change of sperm proteins after cryopreservation may play negative roles for fertilization and early embryo development. Conventionally, cryostraws (CS) and cryovials (CV) are the most widely used cryopreservation carriers (CPCs) for human sperm cryopreservation accompanied with the use of egg yolk free commercial media. However, the influence of cryopreservation on the proteomic profile of human sperm preserved with the two CPCs is unknown. Therefore the purpose of the present study was to compare the frozen-thawed motility, investigate the proteomic profile of human sperm cryopreserved with the two types of CPCs, and identify the susceptible proteins that play key roles for sperm function and fertility. Methods The present study compared the cryosurvival of human sperm frozen with the two different CPCs and identified the sperm proteomic changes by using the isobaric tags for relative and absolute quantification labeling technique coupled with 2D LC–MS/MS analysis after freezing and thawing. Results Our results indicated that sperm cryopreserved with CV showed higher values for percentage of motile sperm and forward activity rate than those with CS. Compared to fresh sperm, 434 and 432 proteins were differentially identified in human sperm cryopreserved with CS and CV, respectively. Conclusion The proteomic profiles of human sperm are greatly affected by cryopreservation with either type of CPC. GO analysis revealed that most of the differentially identified sperm proteins enriched in the extracellular membrane-bounded organelles, cytoplasm and cytosol. In addition, 106 susceptible proteins having known identities related to sperm functions were identified. In general, cryovial seems to be the preferred CPC for human sperm cryopreservation based on the post-thaw motility parameters and the effect on sperm proteomic profiles. These results are beneficial for the insight into the understanding of the cryoinjury mechanism of sperm and the development of human sperm cryopreservation strategies. Electronic supplementary material The online version of this article (10.1186/s12014-019-9244-2) contains supplementary material, which is available to authorized users.

Unraveling Subcellular and Ultrastructural Changes During Vitrification of Human Spermatozoa: Effect of a Mitochondria-Targeted Antioxidant and a Permeable Cryoprotectant

“…Therefore, we used label-free quantitative proteomics analysis to reveal the key proteins altered during the vitrification process. Earlier, proteomics approaches were used to study the alterations in proteins after cryopreservation of human sperm using slow-freezing methods (Wang et al, 2014;Bogle et al, 2017;Fu et al, 2019;Li et al, 2019). Wang et.al (2014) found only 27 proteins that differed in abundance between fresh and cryopreserved sperm due to using lowsensitivity two-dimensional polyacrylamide gel electrophoresis (2-DE).…”

“…Also, subcellular reasons-like changes in the functional proteinsassociated with the fertilization failure of the spermatozoa cannot be clearly explained with conventional semen analysis (Panner Selvam et al, 2019). The damage of spermatozoa proteins during spermatozoa cryopreservation undesirably affects fertilization and early embryonic development (Wang et al, 2014;Bogle et al, 2017;Gholami et al, 2018;Li et al, 2019). Recently, spermatozoa vitrification has been established as an alternative to the slowfreezing method of human spermatozoa.…”

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

“…Such techniques include tandem mass tags (TMTs), isobaric tag for relative and absolute quantitation (iTRAQ) labelling, stable isotope labeling by amino acids in cell culture (SILAC) and isotope-coded affinity tag (ICAT). Among these techniques, iTRAQ labelling is widely used in sperm proteomics [27,28]. Recently, an alternative label-free technique was developed to profile the proteins in sperm cells.…”

Anatomic characteristics of epididymis based on histology, proteomic, and 3D reconstruction

“…However, in fact, the concentration of sp24666 was found to increase significantly from Cor 5 to Cor 6 in our study. P13611‐associated proteins were highly expressed in spermatozoa that recognized the zona pellucida, 26 and the protein GO: 0009987 was highly expressed in highly active spermatozoa 27 . These proteins only increased 10‐fold in the initial stage of the vas deferens transition from the cauda and were not highly expressed in the caput, corpus, and cauda.…”