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Cytochemical Reactions of Bovine Spermatozoa and Seminal Plasma

Departments of Zoology and Entomology and Animal Husbandry, Iowa State College, Ames¹

ABSTRACT

By using accepted cytochemical techniques the following reactions were observed in bovine spermatozoa: Acid phosphatase was present throughout with a slight reaction in the equatorial segment when sodium glycerophosphate was used as a substrate at pH 5.0. Alkaline phosphatase was present, with the exception of the neck, when the same substrate was employed at pH 9.0. Enzymatic activity was found in the neck, middle-piece and tail when adenosine triphosphate was used at pH 5.0. The occurrence of an enzyme acting upon hexose diphosphate (fructose-l,6-diphosphate) at pH 9.0 was noted in the head, neck, middle-piece and tail. Gram's stain and the Feulgen reaction were positive throughout the head, indicating the presence of both ribonucleic and desoxy-ribonucleic acids. Metachromatic staining was not obtained with toluidin blue, indicating the absence of mucopolysaccarides. Fat, including phospholipid, was localized in the middle-piece. The acrosome was characterized by a slight positive reaction with the periodic acid-Schiff reaction which was not due to glycogen, glycolipid or mucopolysaccharide. Furthermore, the protoplasmic droplet which occasionally was present on the tail gave an intense reaction with the periodic acid-Schiff procedure.

Seminal plasma gave the following reactions: Acid phosphatase was positive with sodium glycerophosphate as a substrate at pH 5.0 and alkaline phosphatase was characterized by a weak response with hexose diphosphate (fructose-1,6-diphosphate) at pH 9.0. Gram's stain and toluidin blue gave no reactions. Baker's acid hematein test for phospholipid was negative, but the control preparations were characterized by a bluish-grey stippling and a positive test was obtained with Sudan black B. No reaction was observed with Bodian's protargol method. A slight positive reaction was obtained with the periodic acid-Schiff reaction.

This project is supported in part by funds furnished by Dairy Genetics, Des Moines, Ia.; Eastern Iowa Artificial Breeding Association, Cedar Rapids, Ia.; and Northwest Iowa Federated Breeders Coop., Sheldon, Ia. Project 936. Journal paper no. J-1967.