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Proteomic approaches for profiling negative fertility markers in inferior boar spermatozoa.

Kwon WS, Oh SA, Kim YJ, Rahman MS, Park YJ, Pang MG - Sci Rep (2015)

Bottom Line: Nineteen of these proteins exhibited decreased expression in large litter size samples and increased expression in the small litter group.We then identified signaling pathways associated with the differentially expressed protein markers.Glutathione S-transferase Mu3 and glutathione peroxidase 4 were related to the glutathione metabolic pathway and arginine vasopressin receptor 2 was linked to vasopressin R2/STAT.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal Science and Technology, Chung-Ang University, Anseong, Gyeonggi-do 456-756, Korea.

ABSTRACT
The ability to predict male fertility is of paramount importance for animal breeding industries and for human reproduction. Conventional semen analysis generally provides information on the quantitative parameters of spermatozoa, but yields no information concerning its functional competence. Proteomics have identified candidates for male fertility biomarkers, but no studies have clearly identified the relationship between the proteome and sperm fertility. Therefore, we performed a proteomic analysis to investigate small and large litter size boar spermatozoa and identify proteins related to male fertility. In this study, 20 proteins showed differential expression levels in small and large litter size groups. Nineteen of these proteins exhibited decreased expression in large litter size samples and increased expression in the small litter group. Interestingly, only one protein was highly expressed in the large litter size spermatozoa. We then identified signaling pathways associated with the differentially expressed protein markers. Glutathione S-transferase Mu3 and glutathione peroxidase 4 were related to the glutathione metabolic pathway and arginine vasopressin receptor 2 was linked to vasopressin R2/STAT. In summary, this is the first study to consider negative fertility biomarkers, and the identified proteins could potentially be used as biomarkers for the detection of inferior male fertility.

No MeSH data available.


Related in: MedlinePlus

Differentially expressed spots in small and large litter size spermatozoa by 2-D electrophoretic separation.Spots in boxes (upper panels) indicate individual spots from 3 replicates of small and large litter size spermatozoa. The symbols indicate replicate and litter size (S = Small litter size, L = Large litter size, 1 = First replicate gel, 2 = Second replicate gel, 3 = Third replicate gel). Graphs (lower panels) indicate spot densities. Data represent mean ± SEM, n = 3. Protein expression ratios denoted with an asterisk were significantly different (*P < 0.05).
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f1: Differentially expressed spots in small and large litter size spermatozoa by 2-D electrophoretic separation.Spots in boxes (upper panels) indicate individual spots from 3 replicates of small and large litter size spermatozoa. The symbols indicate replicate and litter size (S = Small litter size, L = Large litter size, 1 = First replicate gel, 2 = Second replicate gel, 3 = Third replicate gel). Graphs (lower panels) indicate spot densities. Data represent mean ± SEM, n = 3. Protein expression ratios denoted with an asterisk were significantly different (*P < 0.05).

Mentions: To analyze differences in male fertility at the protein level, we compared the protein expression profiles of spermatozoa from boars that yielded small and large litter sizes. 2-DE analysis revealed differences among proteins between low and high fertility groups (Fig. 1, Supplementary Fig. S1). It was important to assess the level of variation in the 2-DE evaluations of boar spermatozoa. To address this issue, detailed gel analyses were performed using the SigmaGel software (Jandel Scientific, San Rafael, CA) on three paired experiments (Fig. 1, Supplementary Fig. S1). Figure 1 depicts the 2-DE gels of spermatozoa from small and large litter size samples and illustrates that the majority of the protein spots were significantly different between the groups (>4-fold). We identified 357 ± 21 and 328 ± 17 protein spots in spermatozoa from small and large litter size groups, respectively. Because the standard errors of protein spot numbers for each group were not high, we suggest that possible variations have been minimized. Approximately 102 protein spots were consistently observed in each sample. Of these 102 spots, 25 spots showed differential expression in intensity between small and large litter size groups. There were 2 spots with decreased expression in the small litter size sample and increased expression in the large size group; 23 spots were decreased in the large litter size sample but increased in the small size group. One of the two proteins highly expressed in large litter size sperm, and 19 of the 23 proteins highly expressed in small litter size sperm, were identified by LC-MS/MS analysis (spots no. 1, 11, 15, 19, and 24 were not identified, Table 1).


Proteomic approaches for profiling negative fertility markers in inferior boar spermatozoa.

Kwon WS, Oh SA, Kim YJ, Rahman MS, Park YJ, Pang MG - Sci Rep (2015)

Differentially expressed spots in small and large litter size spermatozoa by 2-D electrophoretic separation.Spots in boxes (upper panels) indicate individual spots from 3 replicates of small and large litter size spermatozoa. The symbols indicate replicate and litter size (S = Small litter size, L = Large litter size, 1 = First replicate gel, 2 = Second replicate gel, 3 = Third replicate gel). Graphs (lower panels) indicate spot densities. Data represent mean ± SEM, n = 3. Protein expression ratios denoted with an asterisk were significantly different (*P < 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4562270&req=5

f1: Differentially expressed spots in small and large litter size spermatozoa by 2-D electrophoretic separation.Spots in boxes (upper panels) indicate individual spots from 3 replicates of small and large litter size spermatozoa. The symbols indicate replicate and litter size (S = Small litter size, L = Large litter size, 1 = First replicate gel, 2 = Second replicate gel, 3 = Third replicate gel). Graphs (lower panels) indicate spot densities. Data represent mean ± SEM, n = 3. Protein expression ratios denoted with an asterisk were significantly different (*P < 0.05).
Mentions: To analyze differences in male fertility at the protein level, we compared the protein expression profiles of spermatozoa from boars that yielded small and large litter sizes. 2-DE analysis revealed differences among proteins between low and high fertility groups (Fig. 1, Supplementary Fig. S1). It was important to assess the level of variation in the 2-DE evaluations of boar spermatozoa. To address this issue, detailed gel analyses were performed using the SigmaGel software (Jandel Scientific, San Rafael, CA) on three paired experiments (Fig. 1, Supplementary Fig. S1). Figure 1 depicts the 2-DE gels of spermatozoa from small and large litter size samples and illustrates that the majority of the protein spots were significantly different between the groups (>4-fold). We identified 357 ± 21 and 328 ± 17 protein spots in spermatozoa from small and large litter size groups, respectively. Because the standard errors of protein spot numbers for each group were not high, we suggest that possible variations have been minimized. Approximately 102 protein spots were consistently observed in each sample. Of these 102 spots, 25 spots showed differential expression in intensity between small and large litter size groups. There were 2 spots with decreased expression in the small litter size sample and increased expression in the large size group; 23 spots were decreased in the large litter size sample but increased in the small size group. One of the two proteins highly expressed in large litter size sperm, and 19 of the 23 proteins highly expressed in small litter size sperm, were identified by LC-MS/MS analysis (spots no. 1, 11, 15, 19, and 24 were not identified, Table 1).

Bottom Line: Nineteen of these proteins exhibited decreased expression in large litter size samples and increased expression in the small litter group.We then identified signaling pathways associated with the differentially expressed protein markers.Glutathione S-transferase Mu3 and glutathione peroxidase 4 were related to the glutathione metabolic pathway and arginine vasopressin receptor 2 was linked to vasopressin R2/STAT.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal Science and Technology, Chung-Ang University, Anseong, Gyeonggi-do 456-756, Korea.

ABSTRACT
The ability to predict male fertility is of paramount importance for animal breeding industries and for human reproduction. Conventional semen analysis generally provides information on the quantitative parameters of spermatozoa, but yields no information concerning its functional competence. Proteomics have identified candidates for male fertility biomarkers, but no studies have clearly identified the relationship between the proteome and sperm fertility. Therefore, we performed a proteomic analysis to investigate small and large litter size boar spermatozoa and identify proteins related to male fertility. In this study, 20 proteins showed differential expression levels in small and large litter size groups. Nineteen of these proteins exhibited decreased expression in large litter size samples and increased expression in the small litter group. Interestingly, only one protein was highly expressed in the large litter size spermatozoa. We then identified signaling pathways associated with the differentially expressed protein markers. Glutathione S-transferase Mu3 and glutathione peroxidase 4 were related to the glutathione metabolic pathway and arginine vasopressin receptor 2 was linked to vasopressin R2/STAT. In summary, this is the first study to consider negative fertility biomarkers, and the identified proteins could potentially be used as biomarkers for the detection of inferior male fertility.

No MeSH data available.


Related in: MedlinePlus