Abstract
BackgroundPoor semen quality is one of the main causes of infertility. We have generated a set of monoclonal antibodies to human sperm and used them to investigate sperm quality. Some of these antibodies found differences in the expression of proteins between normal sperm and pathological sperm displaying severe defects. One of them was the Hs-14 antibody.The aim of this paper was to determine the target protein of the Hs-14 monoclonal antibody and to investigate the expression of the Hs-14-reacting protein on the sperm of asthenozoospermic men with sperm motility defect and of healthy normozoospermic men.MethodsIndirect immunofluorescence, one-dimensional and two-dimensional polyacrylamide gel electrophoresis, immunoblotting and mass spectrometry.ResultsThe Hs-14 antibody binds fibronectin, β-tubulin and valosin-containing protein - new name for this protein is transitional endoplasmic reticulum ATPase (TERA). Since the Hs-14 reaction with TERA remained the strongest at the highest antibody dilution, and Hs-14 consistently labelled the same spot or band as the monospecific anti-TERA antibody on immunoblots, we assume that TERA is an Hs-14-specific protein. Binding of fibronectin and β-tubulin might represent nonspecific cross-reactivity or Hs-14 reaction with similar epitopes of these proteins.A significant difference (P < 0.001) in immunofluorescence staining with Hs-14 was found between the normozoospermic and asthenozoospermic men.ConclusionThe Hs-14 antibody enables discrimination between sterile or subfertile asthenozoospermic and fertile normozoospermic men. Decreased levels of TERA in men can be used as a biomarker of reduced fertility.Electronic supplementary materialThe online version of this article (doi:10.1186/s12610-015-0025-0) contains supplementary material, which is available to authorized users.
Highlights
La pauvre qualité de la semence est l’une des causes d’infertilité
L’anticorps Hs-14 permet de différencier les hommes stériles ou subfertiles asthénozoospermiques des hommes fertiles normozoospermiques
Our antibodies Hs-8, Hs-14 and Hs-36 reliably bound to the acrosomes of spermatozoa in normozoospermic men (60–80 % cells labelled), while their binding was lower in ejaculates with pathological spermiograms (30–40 % cells labelled) [8]
Summary
La pauvre qualité de la semence est l’une des causes d’infertilité. Nous avons généré une série d’anticorps monoclonaux contre le sperme humain et nous l’avons utilisée pour examiner la qualité du sperme. We have generated a set of monoclonal antibodies to human sperm and used them to investigate sperm quality Some of these antibodies found differences in the expression of proteins between normal sperm and pathological sperm displaying severe defects. Using the set of monoclonal antibodies that we generated in our laboratory we were able to perform a systematic and continuous analysis of ejaculates for the needs of assisted reproduction These antibodies can detect poor quality sperm samples even in cases when the parameters of ejaculates meet the requirements of WHO classification for normozoospermics. The ability of our anti-acrosomal antibodies to recognize defective spermatozoa was confirmed by assessment of sperm in the mice that were exposed to pollutants [9, 10] Both findings undoubtedly suggest the importance of the detected proteins in fertilization. It is obvious that sperm testing must be comprehensive—optimally using a panel of antibodies
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