Objective: To examine the effect of epidermal growth factor (EGF) on human sperm capacitation and the involvement of protein phosphorylation in the regulation of the EGF action.
Design: Human sperm were capacitated in modified Krebs-Ringer's bicarbonate medium of Biggers, Whitten, and Whittingham in the presence of EGF and various agents known to phosphorylate the EGF receptor. The chlortetracycline fluorescence assay was used to monitor capacitated sperm. Capacitation was confirmed by the ability of sperm to undergo the acrosome reaction in response to solubilized mouse zonae pellucidae (ZP).
Results: In 15 minutes, the appearance of the clear perimeter pattern increased significantly in the sperm treated with 100 ng/mL EGF compared with the controls. In EGF-treated sperm, the percent clear perimeter pattern remained stable for 3 hours without affecting the acrosome reaction pattern and the motility. Epidermal growth factor stimulated the appearance of the clear perimeter pattern at concentrations > 100 pg/mL. The stimulation by EGF was attenuated by the treatment with genistein, 12-O-tetradecanoylphorbol-13-acetate, or thapsigargin. In sperm that were incubated in the presence of 100 ng/mL EGF for 30 minutes and further induced the acrosome reaction by mouse ZP, the percent acrosome reaction pattern increased significantly compared with the controls.
Conclusion: Epidermal growth factor stimulates human sperm capacitation by activating the tyrosine kinase of the EGF receptor which is regulated by multisite phosphorylation.