Maturational changes of the CD52-like epididymal glycoprotein on cynomolgus monkey sperm and their apparent reversal in capacitation conditions

• Published in: Molecular reproduction and development Vol. 57; no. 3; pp. 280 - 289
• Main Authors: Yeung, Ching-Hei, Schröter, Sabine, Kirchhoff, Christiane, Cooper, Trevor G.
• Format: Journal Article
• Language: English
• Published: New York John Wiley & Sons, Inc 01.11.2000; Wiley-Liss
• Subjects: Alemtuzumab; alpha-Fetoproteins - pharmacology; Animals; Antibodies, Monoclonal; Antibodies, Monoclonal, Humanized; Antibodies, Neoplasm; Antigens, CD - metabolism; Antigens, Neoplasm; Biological and medical sciences; Blotting, Western; CD52 Antigen; Cell differentiation, maturation, development, hematopoiesis; Cell physiology; decapacitation; epididymal secretion; Epididymis - metabolism; Fundamental and applied biological sciences. Psychology; Glycoproteins - metabolism; HE5; Macaca fascicularis; Male; Molecular and cellular biology; N-Acetylneuraminic Acid - metabolism; Neuraminidase - metabolism; Protease Inhibitors - pharmacology; Salts; Sperm Capacitation - physiology; sperm maturation; Sperm Maturation - physiology; sperm protein; Spermatozoa - metabolism; Spermatozoa - physiology; Staining and Labeling; Temperature; Flow cytometry; Secretion; Glycoprotein; Monkey; Male genital duct; Fecundation; Male genital system; Ripening; Reproduction; Vertebrata; Spermatozoid capacitation; Epididymis; Mammalia; Primates; Immunoblotting assay
• ISSN: 1040-452X; 1098-2795
• DOI: 10.1002/1098-2795(200011)57:3<280::AID-MRD10>3.0.CO;2-5

A major epididymal secretory protein in men has a colinear cDNA sequence with lymphocyte CD52, a sialylated glycoprotein. Immunostaining and flow cytometric detection of cynomolgus monkey sperm CD52 during epididymal maturation showed increases from 20 to 85% stained sperm from the caput to the corpus with staining intensities doubled. Freshly prepared cauda sperm showed only 10% staining while they markedly increased in percentage and intensity of staining upon incubation at 37°C under capacitating conditions, but not at 4°C. Western blotting of proteins from fresh cauda sperm revealed no less antigen than corpus sperm. Staining of ejaculated sperm exhibited similar increases during incubation. Further washing with a high salt medium before staining to remove any electrostatically‐bound molecules masking the antigen showed no effect. Incubation‐induced increases in antigen binding were accelerated by the addition of neuraminidase (0.25 and 0.5 U/ml), but not affected by the sialyl residue‐rich fetuin (5 mg/ml) competing for any endogenous neuraminidase. There were no concomitant decreases in the staining of sialic acid residues during capacitation‐incubation. These findings suggest a cryptic antigen epitope site as a consequence of sperm maturation and subsequent re‐exposure under capacitation conditions, but not due to the removal of sialic acid residues by endogenous neuraminidase. Involvement of endogenous proteases was also ruled out, as incubation in the presence of protease inhibitors did not hinder the increases but resulted in a dose‐dependent enhancement in staining, suggesting some protease‐sensitive unmasking process. In conclusion, the monkey epididymal secreted CD52 on sperm underwent changes in antigenic characteristics during sperm maturation which were reversed under capacitation conditions. Mol. Reprod. Dev. 57:280–289, 2000. © 2000 Wiley‐Liss, Inc.