Ribonucleic acid synthesis by spermatozoa from the rat and hamster

Abstract

Spermatozoa from the cauda of the epididymis of the hamster and rat were incubated with [5 3H]uridine and glucose. By using a procedure avoiding bacterial and other cellular contamination, sonic extracts were prepared and digested with deoxyribonuclease and Pronase. Radioactive RNA of high molecular weight was isolated by two methods: gel filtration on Sephadex G 75 columns and polyacrylamide gel electrophoresis in which it migrated in the region of 28S and 23S RNA markers. The macromolecules were alkali labile and hydrolysed by ribonuclease. From 3H radioactivity and E260 of the isolated RNA the rate of incorporation of uridine into RNA of spermatozoa was calculated to be 0.1-0.5 nmol/h per mg of RNA.