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Major protein alterations in spermatozoa from infertile men with unilateral varicocele.

Agarwal A, Sharma R, Durairajanayagam D, Ayaz A, Cui Z, Willard B, Gopalan B, Sabanegh E - Reprod. Biol. Endocrinol. (2015)

Bottom Line: Proteins were extracted and separated by 1-D SDS-PAGE.Bands were digested and identified on a LTQ-Orbitrap Elite hybrid mass spectrometer system.Bioinformatic analysis identified the pathways and functions of the differentially expressed proteins (DEP).

View Article: PubMed Central - PubMed

Affiliation: Center for Reproductive Medicine, Glickman Urological & Kidney Institute, Cleveland Clinic, Mail Code X-11, 10681 Carnegie Avenue, Cleveland, OH, 44195, USA. agarwaa@ccf.org.

ABSTRACT

Background: The etiology of varicocele, a common cause of male factor infertility, remains unclear. Proteomic changes responsible for the underlying pathology of unilateral varicocele have not been evaluated. The objective of this prospective study was to employ proteomic techniques and bioinformatic tools to identify and analyze proteins of interest in infertile men with unilateral varicocele.

Methods: Spermatozoa from infertile men with unilateral varicocele (n=5) and from fertile men (control; n=5) were pooled in two groups respectively. Proteins were extracted and separated by 1-D SDS-PAGE. Bands were digested and identified on a LTQ-Orbitrap Elite hybrid mass spectrometer system. Bioinformatic analysis identified the pathways and functions of the differentially expressed proteins (DEP).

Results: Sperm concentration, motility and morphology were lower, and reactive oxygen species levels were higher in unilateral varicocele patients compared to healthy controls. The total number of proteins identified were 1055, 1010 and 1042 in the fertile group, and 795, 713 and 763 proteins in the unilateral varicocele group. Of the 369 DEP between both groups, 120 proteins were unique to the fertile group and 38 proteins were unique to the unilateral varicocele group. Compared to the control group, 114 proteins were overexpressed while 97 proteins were underexpressed in the unilateral varicocele group. We have identified 29 proteins of interest that are involved in spermatogenesis and other fundamental reproductive events such as sperm maturation, acquisition of sperm motility, hyperactivation, capacitation, acrosome reaction and fertilization. The major functional pathways of the 359 DEP related to the unilateral varicocele group involve metabolism, disease, immune system, gene expression, signal transduction and apoptosis. Functional annotations showed that unilateral varicocele mostly affected small molecule biochemistry and post-translational modification proteins. Proteins expressed uniquely in the unilateral varicocele group were cysteine-rich secretory protein 2 precursor (CRISP2) and arginase-2 (ARG2).

Conclusions: The expression of these proteins of interest are altered and possibly functionally compromised in infertile men with unilateral varicocele. If validated, these proteins may lead to potential biomarker(s) and help better understand the mechanism involved in the pathophysiology of unilateral varicocele in infertile men.

No MeSH data available.


Related in: MedlinePlus

Top disease and function networks and involvement of differentially expressed proteins in free radical scavenging, neurological disease, skeletal and muscular disorder. Green color shows that these differentially expressed proteins (DEPs) were underexpressed and red shows overexpression of DEPs in unilateral varicocele group compared to the fertile group. The gradation of color reflects their intensity/ abundance of expression. Gradation of color reflects their intensity/ abundance of expression (e.g. brighter the red, the larger the protein expression).
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Fig7: Top disease and function networks and involvement of differentially expressed proteins in free radical scavenging, neurological disease, skeletal and muscular disorder. Green color shows that these differentially expressed proteins (DEPs) were underexpressed and red shows overexpression of DEPs in unilateral varicocele group compared to the fertile group. The gradation of color reflects their intensity/ abundance of expression. Gradation of color reflects their intensity/ abundance of expression (e.g. brighter the red, the larger the protein expression).

Mentions: The top networks involving the DEPs, identified by IPA and DAVID software functional annotations, are shown in Table 4. Networks of interactions of some of the DEP identified in our study and their possible roles in the various functions as well as their subcellular localization are shown in Figures 5, 6 and 7.Table 4


Major protein alterations in spermatozoa from infertile men with unilateral varicocele.

Agarwal A, Sharma R, Durairajanayagam D, Ayaz A, Cui Z, Willard B, Gopalan B, Sabanegh E - Reprod. Biol. Endocrinol. (2015)

Top disease and function networks and involvement of differentially expressed proteins in free radical scavenging, neurological disease, skeletal and muscular disorder. Green color shows that these differentially expressed proteins (DEPs) were underexpressed and red shows overexpression of DEPs in unilateral varicocele group compared to the fertile group. The gradation of color reflects their intensity/ abundance of expression. Gradation of color reflects their intensity/ abundance of expression (e.g. brighter the red, the larger the protein expression).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4383193&req=5

Fig7: Top disease and function networks and involvement of differentially expressed proteins in free radical scavenging, neurological disease, skeletal and muscular disorder. Green color shows that these differentially expressed proteins (DEPs) were underexpressed and red shows overexpression of DEPs in unilateral varicocele group compared to the fertile group. The gradation of color reflects their intensity/ abundance of expression. Gradation of color reflects their intensity/ abundance of expression (e.g. brighter the red, the larger the protein expression).
Mentions: The top networks involving the DEPs, identified by IPA and DAVID software functional annotations, are shown in Table 4. Networks of interactions of some of the DEP identified in our study and their possible roles in the various functions as well as their subcellular localization are shown in Figures 5, 6 and 7.Table 4

Bottom Line: Proteins were extracted and separated by 1-D SDS-PAGE.Bands were digested and identified on a LTQ-Orbitrap Elite hybrid mass spectrometer system.Bioinformatic analysis identified the pathways and functions of the differentially expressed proteins (DEP).

View Article: PubMed Central - PubMed

Affiliation: Center for Reproductive Medicine, Glickman Urological & Kidney Institute, Cleveland Clinic, Mail Code X-11, 10681 Carnegie Avenue, Cleveland, OH, 44195, USA. agarwaa@ccf.org.

ABSTRACT

Background: The etiology of varicocele, a common cause of male factor infertility, remains unclear. Proteomic changes responsible for the underlying pathology of unilateral varicocele have not been evaluated. The objective of this prospective study was to employ proteomic techniques and bioinformatic tools to identify and analyze proteins of interest in infertile men with unilateral varicocele.

Methods: Spermatozoa from infertile men with unilateral varicocele (n=5) and from fertile men (control; n=5) were pooled in two groups respectively. Proteins were extracted and separated by 1-D SDS-PAGE. Bands were digested and identified on a LTQ-Orbitrap Elite hybrid mass spectrometer system. Bioinformatic analysis identified the pathways and functions of the differentially expressed proteins (DEP).

Results: Sperm concentration, motility and morphology were lower, and reactive oxygen species levels were higher in unilateral varicocele patients compared to healthy controls. The total number of proteins identified were 1055, 1010 and 1042 in the fertile group, and 795, 713 and 763 proteins in the unilateral varicocele group. Of the 369 DEP between both groups, 120 proteins were unique to the fertile group and 38 proteins were unique to the unilateral varicocele group. Compared to the control group, 114 proteins were overexpressed while 97 proteins were underexpressed in the unilateral varicocele group. We have identified 29 proteins of interest that are involved in spermatogenesis and other fundamental reproductive events such as sperm maturation, acquisition of sperm motility, hyperactivation, capacitation, acrosome reaction and fertilization. The major functional pathways of the 359 DEP related to the unilateral varicocele group involve metabolism, disease, immune system, gene expression, signal transduction and apoptosis. Functional annotations showed that unilateral varicocele mostly affected small molecule biochemistry and post-translational modification proteins. Proteins expressed uniquely in the unilateral varicocele group were cysteine-rich secretory protein 2 precursor (CRISP2) and arginase-2 (ARG2).

Conclusions: The expression of these proteins of interest are altered and possibly functionally compromised in infertile men with unilateral varicocele. If validated, these proteins may lead to potential biomarker(s) and help better understand the mechanism involved in the pathophysiology of unilateral varicocele in infertile men.

No MeSH data available.


Related in: MedlinePlus