Polyamine profiles in rat testis, germ cells and Sertoli cells during testicular maturation

Polyamine cellular concentrations (putrescine, spermidine and spermine) in the rat testis and testicular cell types were determined by fluorescence spectroscopy of their dansyl derivatives. A method is described to separate dansylated polyamines by high performance liquid chromatography in less than 12 minutes. In rat Sertoli cells, polyamine concentrations (per mg DNA) were greater than those in germ cells and the testis. The concentrations of all three polyamines increased with age. Concentrations of spermidine and spermine in germ cells also increased with age and leveled off after 27 to 35 days. On the other hand, higher putrescine levels were found in the testis of young rats (13 to 22 days) while the greatest spermidine and spermine contents were observed in the testis from rats of 31 to 35 days old. Of great interest, Sertoli cells from all age groups studied released a relatively large quantity of putrescine and a smaller amount of spermidine, but no spermine, into culture media. The amount of polyamine released by Sertoli cells varied with the age of the animal. Sertoli cells from 27-day-old rats released the greatest quantity of putrescine on a per mg DNA basis. The release of putrescine increased after hypotonic treatment that removed contaminating germ cells from the remaining Sertoli cells. It is concluded that cellular polyamine levels in the rat testis, germ cells and cultured Sertoli cells and the amount of polyamines released by Sertoli cells were age-dependent during the first wave of spermatogenesis.