title: Major protein alterations in spermatozoa from infertile men with unilateral varicocele.
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.
authors.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.
authors.forename: Ashok
authors.initials: A
authors.lastname: Agarwal
authors.affiliation: Center for Reproductive Medicine, Glickman Urological & Kidney Institute, Cleveland Clinic, Mail Code X-11, 10681 Carnegie Avenue, Cleveland, OH, 44195, USA. sharmar@ccf.org.
authors.forename: Rakesh
authors.initials: R
authors.lastname: Sharma
authors.affiliation: Center for Reproductive Medicine, Glickman Urological & Kidney Institute, Cleveland Clinic, Mail Code X-11, 10681 Carnegie Avenue, Cleveland, OH, 44195, USA. damayanthi.d@gmail.com.
authors.forename: Damayanthi
authors.initials: D
authors.lastname: Durairajanayagam
authors.affiliation: Center for Reproductive Medicine, Glickman Urological & Kidney Institute, Cleveland Clinic, Mail Code X-11, 10681 Carnegie Avenue, Cleveland, OH, 44195, USA. ahmetayaz@gmail.com.
authors.forename: Ahmet
authors.initials: A
authors.lastname: Ayaz
authors.affiliation: Center for Reproductive Medicine, Glickman Urological & Kidney Institute, Cleveland Clinic, Mail Code X-11, 10681 Carnegie Avenue, Cleveland, OH, 44195, USA. zhihongcui_4@126.com.
authors.forename: Zhihong
authors.initials: Z
authors.lastname: Cui
authors.affiliation: Proteomics Research Core Services, Lerner Research Institute, Cleveland Clinic, Cleveland, OH, 44195, USA. willarb@ccf.org.
authors.forename: Belinda
authors.initials: B
authors.lastname: Willard
authors.affiliation: Proteomics Research Core Services, Lerner Research Institute, Cleveland Clinic, Cleveland, OH, 44195, USA. banugopalan@yahoo.com.
authors.forename: Banu
authors.initials: B
authors.lastname: Gopalan
authors.affiliation: Center for Reproductive Medicine, Glickman Urological & Kidney Institute, Cleveland Clinic, Mail Code X-11, 10681 Carnegie Avenue, Cleveland, OH, 44195, USA. sabanee@ccf.org.
authors.forename: Edmund
authors.initials: E
authors.lastname: Sabanegh
mesh_terms: D000328:Adult; D002853:Chromatography, Liquid; D019295:Computational Biology; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007248:Infertility, Male / Q000150:complications / Q000378:metabolism*; D008297:Male; D013058:Mass Spectrometry; D011446:Prospective Studies; D011506:Proteins / Q000737:chemistry / Q000378:metabolism; D040901:Proteomics; D013094:Spermatozoa / Q000378:metabolism*; D014646:Varicocele / Q000150:complications / Q000378:metabolism*
chemical_list: D011506:Proteins