Determination of Acrosomal Proteolytic Activity in Spermatozoa Collected from Quail Treated with Diethylstilbestrol, Using Gelatin-Substrate Slide Technique

Abstract

The present experiment was designed to ascertain whether gelatin-substrate slide technique devised for detection of the proteolytic activity of acrosomal contents can be used for quail sperm, and to determine whether treatment with the endocrine disrupting chemical, diethylstilbestrol (DES), has deleterious effect on the modulation of acrosomal proteolytic activity (APA) in quail sperm. It was observed on the slide under a phase-contrast microscope that a halo formed around a single spermatozoon resulting from localized proteolytic digestion of gelatin. The mean halo diameters of sperm showed a dose-dependent decrease with increasing levels of trypsin inhibitor, ovomucoid which drastically blocked the halo formation. These findings indicate that the gelatin-substrate slide technique may be a more effective and more simplified method for evaluation of APA in quail sperm. The mean halo diameters were significantly (P<0.05) low (52.2μm) in sperm collected from DES-injected quail compared with those of sperm without injection (86.3μm) implying that treatment with DES provides a decrease of APA in agreement with the acrosomal damage.

Keywords

• acrosomal proteolytic activity (apa)
• diethylstilbestrol
• gelatin-substrate slide
• quail sperm