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RESEARCH ARTICLE

Human seminal fibronectin fragmentation patterns and their domain immunoreactivities in leucocytospermic patients

Ewa M. Kratz A E , Marcin Wójtowicz A , Magdalena Przybysz D , Ricardo Faundez B C and Iwona Kątnik-Prastowska A
+ Author Affiliations
- Author Affiliations

A Department of Chemistry and Immunochemistry, Wrocław Medical University, Bujwida 44a, 50-345 Wrocław, Poland.

B Embryology Laboratory InviMed – European Centre of Motherhood, Rakowiecka 36, 02-532 Warsaw, Poland.

C Department of Large Animal Diseases and Clinic, Division of Animal Reproduction, Andrology and Biotechnology of Reproduction, Faculty of Veterinary Medicine, Warsaw University of Life Sciences, Nowoursynowska 100, 02-797 Warsaw, Poland.

D Deceased. Formerly of Department of Chemistry and Immunochemistry, Wrocław Medical University, Bujwida 44a, 50-345 Wrocław, Poland.

E Corresponding author. Email: ewa.kratz@umed.wroc.pl

Reproduction, Fertility and Development 26(7) 1044-1051 https://doi.org/10.1071/RD13049
Submitted: 14 February 2013  Accepted: 15 July 2013   Published: 7 August 2013

Abstract

The aim of the work was to analyse fibronectin (FN) domain immunoreactivities and profiles of FN fragmentation in seminal plasmas of fertile normozoospermic and infertile leucocytospermic male patients. ELISA with domain-specific monoclonal antibodies and immunoblotting were used in these measurements. Immunoblotting of normal and leucocytospermic seminal plasmas revealed the presence of twelve FN bands of ~70–196 kDa with nearly identical FN profiles under reducing and non-reducing conditions. The epitopes of the cell-, fibrin-, collagen-binding FN domains and the extra domain A (EDA) FN segment retained the ability to bind their specific monoclonal antibodies, whereas the fibrin–heparin domain (N-terminal end) and the area around the disulfide bridges (C-terminal end) of the FN polypeptide did not show any reactivities with their respective specific antibodies. The mean values of cell- (338.4 ± 138.4 and 398.3 ± 310 mg L–1), fibrin- (79.1 ± 38.5 and 145.2 ± 188.8 mg L–1) and collagen-binding (19 ± 19.8 and 50.9 ± 73.4 mg L–1) FN domain immunoreactivities and the relative amount of EDAFN did not show any significant differences between the normal and leucocytospermic groups. The high values of standard deviations for the FN domain immunoreactivities in the leucocytospermic group probably results from different aetiology of leucocytospermia. The profile of FN fragmentation and alterations of FN domain immunoreactivities in seminal plasma may influence their engagement in the fertilisation process. The analysis of seminal FN molecular status would be helpful for selecting the highest quality spermatozoa for use in assisted reproduction techniques.

Additional keywords: fibronectin domains, fibronectin fragments, human seminal plasma, leucocytospermia.


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