Proteomics analysis of intensive exercise-induced disorders of gametogenesis in the testis using isobaric tags for relative and absolute quantification (iTRAQ) analysis

Abstract

In the present study we screened a panel of regulatory proteins associated with gametogenesis disorders in the testis that are induced by intensive exercise. Four-week-old Sprague-Dawley male rats were randomly divided into three groups: a control group, a no-load exercise group and an intensive exercise group. Rats in the control group were free to move in their cage. Rats in the no-load exercise and intensive exercise groups swam for 60minday-1, six times each week, for a total 9-week exercise regimen; rats in the intensive exercise group swam with a load of 6% body mass. After the last exercise session (or at the end of the 9-week period), a sperm count, reproductive hormone assays, histological analysis of the testis and proteomics analysis were performed for each rat. Mean (±s.d.) sperm concentration was significantly lower in the intensive exercise group than in the control and no-load exercise groups (1.36±0.63 vs 2.12±0.53 and 2.57±0.48×106 spermatozoa mL-1 respectively; P<0.05). Serum testosterone concentrations were also significantly lower in the intensive exercise group (P<0.01), whereas gonadotrophin-releasing hormone, LH and FSH concentrations were slightly decreased in the intensive exercise group, but not significantly (P>0.05). Histological analysis showed that the number of spermatogenic cells in the seminiferous tubules was lower in the intensive exercise group than in the control and no-load exercise groups. Proteomics analysis identified 54 proteins that were differentially expressed between the control and intensive exercise groups (31 downregulated, 23 upregulated). Pathway enrichment analysis showed that ribosome and extracellular matrix-receptor interaction pathways play an important role in the signal transduction of testicular gametogenic disorders. Four differentially expressed proteins that were involved in the regulation of reproduction were identified by bioinformatics analysis and validated by targeted mass spectrometry analysis, namely vimentin, collagen α-1(I) chain, fatty acid-binding protein 9 and 40S ribosomal protein S3a. The data suggest that changes in the abundance of differentially expressed proteins after long-term intensive exercise affect the cycle and progression of spermatogenesis, resulting in spermatogenic disorders.

Proteomics-Based Detection of Immune Dysfunction in an Elite Adventure Athlete Trekking Across the Antarctica

“…Targeted proteomics procedures were conducted on DBS samples that were collected before and after, and twice daily during RAAM, and revealed large fold increases for specific immune-related proteins including complement component C7 (359%), complement C4b (231%), serum amyloid A4 protein (210%), inter-alpha-trypsin inhibitor heavy chain H4 (191%), and alpha-1-antitrypsin (188%). These data are consistent with results from multiple human and animal studies showing that immune-related proteins represent a large proportion of those generated during intensive and prolonged acute and chronic exercise training [8][9][10][11][12][13]. These proteins, when combined with psychological, performance, and nutrition intake data, can serve as useful NFOR and OTS biomarkers of immune dysfunction, training distress, exercise-induced muscle damage and exhaustion, and impaired performance capacity [14][15][16][17][18][19][20][21].…”

Chronic exercise training attenuates prostate cancer-induced molecular remodelling in the testis

“…The influence of EX on testis function also depends upon the type and intensity of the EX program. While high-intensity EX was found to be associated with reduced luteinizing hormone (LH), follicle-stimulating hormone (FSH), testosterone and semen parameters, for moderate-intensity EX there was no consensus among the published studies [19][20][21][22]. In subjects with testicular functional impairment caused by aging, obesity or drug treatment, EX was reported to prevent or counteract the decrease in testosterone serum levels and sperm counts [23][24][25].…”

iTRAQ-based comparative proteomics reveal an enhancing role of PRDX6 in the freezability of Mediterranean buffalo sperm

“…Compared with 2-dimensional SDS-PAGE and label free quanti cation, isobaric tags for relative and absolute quantitation (iTRAQ) has the advantages of robustness to variations in sensitivity and chromatographic reproducibility [9]. iTRAQ-coupled LC-MS/MS technology is currently being used to track various biomarkers in spermatogenesis [10,11], sperm maturation [12], concentration and motility changes of ejaculates [13], cryoinjury [14,15] and fertility [16,17]. The parallel reaction monitoring (PRM) technology is recently developed in the use of targeted mass spectrometry.…”