title: Characteristics of testis-specific phosphoglycerate kinase 2 and its association with human sperm quality.
abstract: OBJECTIVE
  Is there an association between the expression of phosphoglycerate kinase (PGK) 2 in spermatozoa and sperm quality in both elderly men and young asthenozoospermia patients?
  
  
  CONCLUSIONS
  Spermatozoa from elderly men and young asthenozoospermia patients show decreased expression of PGK2, which has a close positive relationship with sperm quality.
  
  
  BACKGROUND
  PGK1 and PGK2 are involved in spermatogenesis and thought to be related to sperm motility. However, limited information is known about their temporal-spatial expression in human spermatogenesis and their relationship with sperm quality.
  
  
  METHODS
  This was a case-control study including 30 healthy young males (aged 28-31 years), 30 elderly men (aged 68-70 years), and 30 asthenozoospermic patients (aged 25-40 years, progressive motility <32%) who donated semen samples. Furthermore, young testes samples were obtained from five fathers (27-33 years old) who had died in car accidents, while aged testes samples were obtained from five elderly fathers (78-82 years old) who were prostate cancer patients.
  
  
  METHODS
  Semen samples from young adults, elderly men and asthenozoospermic patients were prepared, and their parameters were assessed by Computer-Aided Sperm Analysis (CASA). Sperm proteins were extracted for western blot analysis. Immunohistochemistry was used to characterize the cellular localization of PGK1 and PGK2 in testes samples. Sperm immunofluorescence quantification experiments identified the differential expression of PGK1 and PGK2 in sperm from young adults, elderly men and asthenozoospermic patients. Antibodies against PGK1 and PGK2 were used to test their influence on sperm motility and penetration into viscous media. A modified Kremer test using methyl cellulose was adopted to assess sperm function via penetration into viscous media.
  
  
  RESULTS
  Cellular localization analysis showed that PGK1 was mainly expressed in spermatogonia whereas PGK2 was mainly expressed in round spermatids. Expression levels of both PGKs were significantly decreased in the testis with ageing (P < 0.05). Western blot and immunofluorescence quantification showed markedly lower expression of PGK2 (P < 0.05) in sperm from elderly men or asthenozoospermic patients compared sperm from with healthy young men. Sperm functional analysis validated the close relationship between expression of PGK2 and sperm motility (staining percentage, r = 0.60, P < 0.05; intensity, r = 0.59, P < 0.05). Use of an anti-PGK2 antibody on sperm significantly decreased their ability to penetrate into a cervical mucus substitute (P < 0.05).
  
  
  CONCLUSIONS
  Before any clinical applications using PGK2 to assess sperm quality can be developed, more cases should be used to evaluate this approach.
  
  
  CONCLUSIONS
  The study provides new insights into the role of PGKs in male reproduction. The results also indicate that PGK2 is a promising molecular candidate for the assessment of sperm quality and the screening of male contraceptive targets.
  
  
  BACKGROUND
  This work was supported by grants from the National Natural Science Foundation of China (no. 81300533, 81370013 and 81000277) and Shandong Provincial Natural Science Foundation, China (ZR2013HQ002). The authors declare no competing financial interests.
authors.affiliation: Central Laboratory, Yantai Yu Huang Ding Hospital/Qingdao University, Yantai 264000, Shandong, P.R. China.
authors.forename: Xue-Xia
authors.initials: XX
authors.lastname: Liu
authors.affiliation: Department of Otorhinolaryngology Head and Neck Surgery, Yantai Yu Huang Ding Hospital/Qingdao University, Yantai 264000, Shandong, P.R. China.
authors.forename: Hua
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Central Laboratory, Yantai Yu Huang Ding Hospital/Qingdao University, Yantai 264000, Shandong, P.R. China.
authors.forename: Xiao-Fang
authors.initials: XF
authors.lastname: Shen
authors.affiliation: Central Laboratory, Yantai Yu Huang Ding Hospital/Qingdao University, Yantai 264000, Shandong, P.R. China male_reproduction@163.com.
authors.forename: Fu-Jun
authors.initials: FJ
authors.lastname: Liu
authors.affiliation: Central Laboratory, Yantai Yu Huang Ding Hospital/Qingdao University, Yantai 264000, Shandong, P.R. China.
authors.forename: Juan
authors.initials: J
authors.lastname: Liu
authors.affiliation: Reproduction Medical Center, Yantai Yu Huang Ding Hospital/Qingdao University, Yantai 264000, Shandong, P.R. China.
authors.forename: Wen-Juan
authors.initials: WJ
authors.lastname: Wang
mesh_terms: D000328:Adult; D000368:Aged; D053627:Asthenozoospermia / Q000378:metabolism*; D015415:Biomarkers / Q000378:metabolism; D016022:Case-Control Studies; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism*; D008297:Male; D010735:Phosphoglycerate Kinase / Q000378:metabolism*; D055101:Semen Analysis; D013091:Spermatogenesis; D013094:Spermatozoa / Q000502:physiology*; D013737:Testis / Q000378:metabolism*
chemical_list: D015415:Biomarkers; D007527:Isoenzymes; D010735:Phosphoglycerate Kinase; C104317:phosphoglycerate kinase, testis specific
keywords: PGK2; asthenozoospermia; sperm motility; sperm quality; spermatogenesis; testis