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. 2019 Jan 8;20(1):203.
doi: 10.3390/ijms20010203.

Proteomic Signatures Reveal Differences in Stress Response, Antioxidant Defense and Proteasomal Activity in Fertile Men with High Seminal ROS Levels

Tânia R Dias  1   2   3 , Luna Samanta  4   5 , Ashok Agarwal  6 , Peter N Pushparaj  7 , Manesh Kumar Panner Selvam  8 , Rakesh Sharma  9
Affiliations

Proteomic Signatures Reveal Differences in Stress Response, Antioxidant Defense and Proteasomal Activity in Fertile Men with High Seminal ROS Levels

Tânia R Dias et al. Int J Mol Sci. .

Abstract

Elevated levels of reactive oxygen species (ROS) are a major cause of male infertility. However, some men with high seminal ROS levels are still fertile. The main objective of this study was to understand the molecular mechanism(s) responsible for the preservation of fertility in those men. Semen samples from fertile men were divided into two groups: control (n = 10, ROS < 102.2 RLU/s/10⁶ sperm) and ROS+ (n = 10, ROS > 102.2 RLU/s/10⁶ sperm). Proteomic analysis of seminal plasma and spermatozoa was used to identify the differentially expressed proteins (DEPs) between the experimental groups, from which some proteins were validated by Western blot (WB). A total of 44 and 371 DEPs were identified between the study groups in the seminal plasma and spermatozoa, respectively. The identified DEPs were primarily involved in oxidoreductase, endopeptidase inhibitor, and antioxidant activities. We validated by WB the underexpression of NADH:ubiquinone oxidoreductase core subunit S1 (p = 0.01), as well as the overexpression of superoxide dismutase 1 (p = 0.03) and peroxiredoxin 4 (p = 0.04) in spermatozoa of ROS+ group. Our data suggest that fertile men with high ROS levels possess an effective antioxidant defense system that protects sperm proteins, as well as an active proteasomal system for degradation of defective proteins.

Keywords: Western blot; antioxidants; bioinformatics; chemiluminescence; differentially expressed proteins; proteomics; reactive oxygen species; seminal plasma; spermatozoa.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Schematic representation of the results obtained by proteomic analysis: (a) number of proteins identified in the seminal plasma and spermatozoa of fertile men (control) and men with high levels of reactive oxygen species (ROS+), as well as the number of differentially expressed proteins (DEPs) between the experimental groups; (b) expression profile of seminal plasma DEPs; and (c) expression profile of spermatozoa DEPs.
Figure 2
Figure 2
Localization of differentially expressed proteins (DEPs) in: (a) seminal plasma; and (b) spermatozoa. The number of DEPs that were overexpressed (grey), underexpressed (orange), unique to control (blue), and unique to ROS+ (yellow) are shown for seminal plasma and spermatozoa.
Figure 3
Figure 3
Graphical representation of Western blot results and respective representative blots for: (a) seminal plasma; and (b) spermatozoa proteins. Results are expressed as mean ± SEM and were considered significant for p < 0.05.
See this image and copyright information in PMC

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