title: Cytochemical and electrophoretic study of the stallion epididymal glycoproteins.
abstract: It has been suggested that proteins produced in specific regions of the epididymis, mostly androgen dependent glycoproteins, are involved in the sperm maturation process. In the present work, the glycoconjugated distribution pattern and the electrophoretic characteristics of the stallion epididymal proteins were examined using lectin probes. The identification in the luminal fluid of some new proteins, probably synthesized and secreted by the epididymis, is an important initial step to investigate their interaction with the stallion sperm membrane. The binding of FITC-lectins (ConA, WGA, LPA, UEA, RCA, HPA) confirmed the presence of macromolecules containing carbohydrate residues in the epithelial cells with a distribution and relative density that was dependent on the epididymal region analyzed. The epithelium displayed affinity for more than one lectin, indicating diversity in the exposed sugar residues. The electrophoretic pattern of proteins obtained from ductus efferentes, corpus and cauda epididymis differed not only from those of the homologous blood serum, but also among the different epididymal regions. The most prominent bands correspond to 66, 55, 45 and 14 kDa proteins, present in different relative concentrations, in the three analyzed regions. A major band of 36 kDa was observed in the cauda epididymis region. The relative concentrations of protein bands of Mw 45, 36, 32, 20 and 18 kDa were significantly increased towards the distal regions of the ductus. The proteins of Mw 66, 55 and 14 kDa showed a relative higher concentration in the efferent ducts, decreasing to 25-30% in the cauda epididymal regions. The Mw 70, 66, 55, 45, 36, 32, 29, 23, 21, 18 and 14 kDa protein bands gave positive PAS reaction indicating that it corresponds to glycoproteins. Mannose residues were detected in the 70, 66, 55, 45, 36 and 32 kDa proteins. WGA-FITC binds to protein bands of Mw 70, 55, 45, 36, 32, 29, 25 and 24 kDa, suggesting the presence of N-linked glycoproteins. However, based on the resistance to the neuraminidase treatment, we suggest that the stallion epididymis contains both O- and N-glycoconjugates, probably in the N-acetyl O-diacetyl form. Although sperm maturation is an androgen-dependent process, no striking differences were detected in the SDS-PAGE obtained from animals in breeding and non-breeding seasons.
authors.affiliation: Faculty of Medicine, University of Chile, Santiago.
authors.forename: C
authors.initials: C
authors.lastname: Retamal
authors.forename: J
authors.initials: J
authors.lastname: Urzúa
authors.forename: E W
authors.initials: EW
authors.lastname: Alves
authors.forename: M L
authors.initials: ML
authors.lastname: López
mesh_terms: D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D004822:Epididymis / Q000737:chemistry* / Q000166:cytology* / Q000648:ultrastructure; D004847:Epithelial Cells / Q000737:chemistry / Q000166:cytology / Q000648:ultrastructure; D016650:Fluorescein-5-isothiocyanate; D006023:Glycoproteins / Q000032:analysis*; D006736:Horses; D037102:Lectins; D008297:Male; D008970:Molecular Weight
chemical_list: D006023:Glycoproteins; D037102:Lectins; D016650:Fluorescein-5-isothiocyanate