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: (a) gel filtration on Sephadex G-75 columns and (b) 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 0.1–0.5nmol/h per mg of RNA.