Glycocalyx characterisation and glycoprotein expression of Sus domesticus epididymal sperm surface samples
Anna Fàbrega A C , Marta Puigmulé A , Jean-Louis Dacheux B , Sergi Bonet A and Elisabeth Pinart A
+ Author Affiliations
- Author Affiliations
A Biotechnology of Animal and Human Reproduction (TechnoSperm), Department of Biology, Institute of Food and Agricultural Technology, University of Girona, Campus Montilivi, s/n, 17071 Girona, Spain.
B Gametes Males et Fertilité, Physiologie de la reproduction et des comportements, UMR 6175 INRA-CNRS-Université de Tours, 37380 Nouzilly, France.
C Corresponding author. Email: anna.fabrega@udg.edu
Reproduction, Fertility and Development 24(4) 619-630 https://doi.org/10.1071/RD11064
Submitted: 10 March 2011 Accepted: 27 September 2011 Published: 25 November 2011
Abstract
The sperm surface is covered with a dense coating of carbohydrate-rich molecules. Many of these molecules are involved in the acquisition of fertilising ability. In the present study, eight lectins (i.e. Arachis hypogae (peanut) agglutinin (PNA), Lens culimaris (lentil) agglutinin-A (LCA), Pisum sativum (pea) agglutin (PSA), Triticum vulgari (wheat) germ agglutinin (WGA), Helix pomatia agglutinin (HPA), Phaseolus vulgaris (red kidney bean) leucoagglutinin (PHA-L), Glycine max (soybean) agglutinin (SBA) and Ulex europaeus agglutinin I (UEA-I)) were investigated to identify changes in the nature and localisation of glycoproteins in boar spermatozoa migrating along the epididymal duct. Complementary procedures included measurement of global lectin binding over the surface of the viable sperm population by flow cytometry, analysis of lectin localisation on the membrane of individual spermatozoa using fluorescence microscopy and the electrophoretic characterisation of the major sperm surface glycoprotein receptors involved in lectin binding. A significant increase was found in sperm galactose, glucose/mannose and N-acetyl-d-glucosamine residues distally in the epididymis. Moreover, the sperm head, cytoplasmic droplet and midpiece were recognised by most of the lectins tested, whereas only HPA and WGA bound to the principal piece and end piece of the sperm tail. Fourteen sperm surface proteins were observed with different patterns of lectin expression between epididymal regions. The sperm glycocalyx modifications observed in the present study provide an insight into the molecular modifications associated with epididymal maturation, which may be correlated with the degree of maturation of ejaculated spermatozoa.
Additional keywords: boar, epididymis, flow cytometry, immunocytochemistry.
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